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BIOL2P98D3Spring2014
PrinciplesofMicrobiology
Dr.CarolynnEPietrangeliPhD
Lecture1June16,2014
CHSC2P98PrinciplesofMicrobiology
SpringD3June16July182014
INSTRUCTOR: Dr.CarolynnE.Pietrangeli,PhD
CONTACT: email: cpietrangeli@brocku.ca
Office:MCF212
OFFICEHOURS: Monday/Friday12PM1PM
Andbyappointment email
request
FORMAT: Lectures,3hourstwiceweekly
Monday/Friday9AM12PM
AS216
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Laboratories
Schedule:AsperpostingonSakai
LaboratoryDemonstrators
ChristeneCarpenterClelandMCF210
Extension5788ccarpenterclela@brocku.ca
MarkLukewichMCF213
Extension3398mlukewich@brocku.ca
Evaluation
Laboratory 35% SeeLaboratoryManual
MidtermExamination 20% FridayJuly49:00AM10:30AM
Discussion Forum will follow DiscussionForumwillfollow
FinalExamination 20% 2hourduration;Registrarsched
DiscussionForum 25% 5exercises/5markseach
SeeSakaiforInstructions
20%content
5%participation p p
ThereareNOmakeuporsupplemental examinationsinBIOL2P98
ThisappliestoLectureExaminations
(MidtermANDFinalExamination)ANDtothe
LaboratoryExamination
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BIOL2P98SPLaboratorySchedule
LaboratorySchedule
Weekof LaboratoryExerciseTitle
Jun 16 Lab 1 Lab Safety Techniques & Enumeration
LabsareinIH309
Jun16 Lab1 LabSafety,Techniques&Enumeration
Jun23 Lab2 BacterialGrowth&Identification
Jun30 CanadaDayweek NOLABS Assignment2onSakai
Jul07 Lab3 TheControlofMicrobialGrowth
Jul14 LabExam Basedonalllabmaterial
Labsections IH309
Section5 Monday 2:00pm 5:00pm
Section3 Tuesday 10:00am 1:00pm
Section1 Tuesday 2:00pm 5:00pm
Section2 Wednesday 2:00pm 5:00pm
ReadallAppendices&Lab1onSakai
CreateaflowchartforLab1
Bringalabcoat&safetyglasses/goggles,Lab1,
flowchartandPhotographicAtlas
DiscussionForums
OccureachFridayduringlastlecturehour(11AM12PM)
Eachweek,weevaluateacaserelevanttolecture/laboratory
content in Microbiology in assigned Small Groups contentinMicrobiologyinassignedSmallGroups
Eachcasewillbeaccompaniedbysetsofquestionstobediscussed
andworkedonbyeachGroup
EachGroupsubmitsaSINGLEelectronicReport(nopaper
submission)respondingtothecasequestionssetforthatGroup
ELECTRONICReport(nopapersubmission)dueby5PMonthe
T d f th f ll i k ( C O tli ) Tuesday ofthefollowingweek(seeCourseOutline)
TheGroupwillreportspecificcontributionsofeachmember
EachmemberoftheGroupearnstheSAMEGRADEfortheReport
DiscussionForumInstructions willbepostedonSakaibyThursday,
June19 includesGrouplistings
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DiscussionForumParticipation
20%ofthetotalgradeearnedforthe
Discussion Forums (5% of the final grade in DiscussionForums(5%ofthefinalgradein
BIOL2P98)isderivedfromparticipationin
DiscussionForumexercises
Attendancewillberecordedbysigninsheet
atthebeginningofeachDiscussionForum
Di i G ill b k d t d ib DiscussionGroupswillbeaskedtodescribe
thespecificcontributionsofeachmemberto
eachGroupReport
Resources
RequiredTextbook
BrockBiologyofMicroorganisms.FourteenthEdition.MadiganMT,Martinko
JM Bender KS Buckley DH Stahl DA Pearson Boston 2012 JM,BenderKS,BuckleyDH,StahlDA.PearsonBoston2012
Providesabasicreviewofconceptstobediscussedduringthecourse
DoesNOTsubstituteforcourselecturesorDiscussionForumcontent
Textbookchapterswillnotbeassigned;studentswillmatchthelecture
contenttorelevantchaptersinthetextbook
TextbookcontentwillbesupplementedbyreadingsmadeavailableonSakai.
Postingsofadditionalreadingswillbeannouncedduringlecturesandon
Sakai
RecommendedTextbookforLaboratory
APhotographicAtlasfortheMicrobiologyLaboratory.Leboffe MJ,PierceBE
4thEdition.MortonPublishing2011
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AcademicIntegrity
AReflectionofPersonalIntegrity
Manythingsmaybestolenfromusbyother
peopleoverthecourseofourlives
Integrityisonethingwecanstealonlyfrom
ourselves
Integrity
Respecting it Respecting yourself Respectingit=Respectingyourself
Preserveit
Beproudofupholdingit
Microbes(Microorganisms)
Singlecelledorganismsvisibleonlyundermagnification,
classifiedin1/3domains:Bacteria/Archaea /
Eukaryotes.AlsoincludesnonlivingViruses/Prions
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Microbiology =Thestudyofmicroorganisms
andtheirinteractionwithotherlivingorganisms
WhatareWeStudying?
MicrobesandTheirImpactonUS!
Bacteria
Protista
A b t
Viruses
Fungi
Prions
Parasites
Protozoa
l h
Amoebaproteus
Bacterium
Lactobacillussake
Helminths
Arthropods
Fungus
Lentinula edodes
Virus
Retrovirus
Prion
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WhyStudyMicrobiology?
Toelucidatebasiclifeprocesses
Microbes excellentmodelsforunderstanding
cellular processes in unicellular and multicellular cellularprocessesinunicellularand multicellular
organisms
Toapplyfindingsinmicrobestohuman
Medicine
Agriculture
Industry
N i l d f Nationaldefense
Theroleoftheinfinitelysmallinnatureis
infinitelylarge LouisPasteur
Plague:Yersinia pestis Polio:Poliovirus
AIDS:HIV1
Anthrax:Bacillusanthracis
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BroaderreachingEffectsof
MicrobiologyResearch
Molecularbiologyandgenetics TheHumanGenome
Project
HumanTherapeutics
Pharmaceuticalsandbiologics
Genetherapy
Ecology
Agriculture
RecombinantInsulin
g
Livestock
Vegetablecrops
Pestcontrol
WhatWillWeLearn?
BythesuccessfulcompletionofBIOL2P98,wewillbe
ableto: able to:
Listtheprincipalmorphological/ultrastructural,
biochemicalandmolecularfeaturesclassifying
microbesasbacteria,archaea orviruses
Describeselectedpathwaysinmicrobialmetabolism
includingreproduction,energyproduction/utilization g p , gy p /
andbiosynthesis
Explainhowmicrobesareexploitedingenetic
engineering,recombinantandcloningtechnologies
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OurPrimaryLesson:
TheValueofADAPATATION
Adjustmentinbehaviour,physiologyand/or j , p y gy /
structureresultinginameasurablechangein
relationshipwiththeenvironment:Plasticity
Breadth
Mechanisms Mechanisms
Counteradaptation
AdaptationReflects
EnvironmentandGenetics
PhysiologicalAdaptation:changein
geneexpressioninresponsetoenvironmental
change;Example endospore formation
GeneticAdaptation:overgrowthofa
mutanttobecomethepredominanttypein
thepopulation;Example antibioticresistance
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DoesMicrobialAdaptationReallyAffectUs?
MRSASuperbug
Opportunistic infections Opportunisticinfections
Autoimmunity
Tumour formation
MRSA:MethicillinresistantStaphylococcusaureus
OpportunisticPneumocystis jirovici
EpsteinBarrvirusassociatedBurkitts Lymphoma
Microbialvs.HumanAdaptation
OddSparringPartners!
Them:
Rapidreproduction
Rampant,random
mutation
Us: Us:
TheImmuneSystem
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InfectiousDiseasesareResponsiblefor
OnethirdofAllDeathsWorldwide
1/3InfectiousDisease
40%
TB Measles
Pneumonia
54MDeathsGlobally Diarrhea Malaria
All i D l i W ld
40%
AllinDevelopingWorld
India/SubSaharanAfrica
Disproportionatetargetingof
Children
http://www.who.int/mediacentre/factsheets/fs310/en/index2.html
LetsBackUpaMinute
HowdoweknowthatMicroorganisms
Interactwithhumans
Causedisease
??
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SpontaneousGeneration
Spontaneousgeneration: livingorganismscanoriginate
fromnonlivingmatter
Pneuma = vital heat; a property of nonliving things Pneuma vitalheat;apropertyofnonlivingthings
Lifeoriginatingfromputrefyingmatterdependsonthe
relativeamountsofpneuma andthe5elements
FrancescoRedi
16261697
Redis Experiment:1668
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TheLongandWindingRoad:
LinkingMicroorganismstoDisease
Antonie vonLeeuwenhoek
TheMicroscope:Firstobservationanddescriptionofmicroorganisms
Prescott
8thed.Fig.1.11b
7thed.Fig.1.3b.
WhatAboutSpontaneous
GenerationforMicroorganisms?
JohnNeedham(17131781)
Broth with microbes in flasks Boiled Clears Flask opened
TheCreationofLifefromNonlivingMatter
BrothwithmicrobesinflasksBoiledClearsFlaskopened
Result:cloudybroth;microorganismgrowth
Needham1748
Lazzaro Spallanzani (17291799)
BrothwithmicrobesinflasksBoiledClearsFlasksealed
Result:clearbroth;microorganismnegative
Whatdoweconcludefromtheseobservations?
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Steam
Dustandmicroorganismstrappedinbend
Liquid cooled slowly
Extendedtime
Nonsterile liquidpouredintoflask Liquidsterilized
Liquidcooledslowly
Sterilitymaintained
Shorttime
ENOUGH,already!
HerestheProof!
Figure1.16a
2012 Pearson Education, Inc.
Flasktippedtoensurecontact
betweenmicroorganismladendust
andsterileliquid
Sterilitybroken
Microorganismgrowth
LouisPasteur(18221895)
Mouse
becomesill
Injecthealthymouse
Inoculumfrom
cowwith
anthrax
SpontaneousGenerationtoGermTheory
Bacilli+spores
culturedfromspleen
What do we conclude from
these observations?
RobertKoch
18431910
Mouse
becomesill
Injecthealthy
mouse
RobertKoch1876
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KochGivesUsPureMicrobiologicalCultures
PotatoSliceCultures:(L)Fingerswab/(R)Nasalswab
PotatoLactoseUIAgar
MixofSaccharomyces,Brettanomyces
andPediococcus
Deoxycholate citrateagar
KochsPostulates 1884
Howdoweestablishacausalrelationshipbetweena
microorganismandaspecificdisease?
1. The microorganism must be present in every case of the 1. Themicroorganismmustbepresent ineverycaseofthe
disease butabsent fromhealthy potentialhosts
2. Thesuspectedmicroorganismmustbeisolatedand
growninapureculture
3. Thesamediseasemustresultwhentheisolated
microorganismisinoculated intoahealthyhost
4. Thesamemicroorganismmustbeisolated againfrom
thediseasedhost
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CaseInPoint TheSpanishFlu
19181920
3% world population 3%worldpopulation
dead(50M)
27%infected(500M)
CausativeAgent:B.influenzae?(H.influenzae)
Isolate%ofSpanishFluCases
B influenzae 5090% Binfluenzae 50 90%
Streptococci 76%
Pneumococci 61%
______________________________________________________________
MultiplevariedstrainsofBinfluenzae wereisolated
fromSpanishfluvictims
i fl i l d f h l h Binfluenzae wasisolatedfrommanyhealthy
individuals
VaccinationwithchloroformkilledBinfluenzae didnot
reproduciblyprotectagainstinfluenza
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RelevanceofKochsPostulatesToday
Goldstandardinmedicalmicrobiology gy
Requireanimalmodelsthatfaithfullyreflect
humandisease
Diseasesnottestable:cholera/rickettsias /
chlamydias
LessonsofHistoryin
MicrobiologyResearch TakingthePrize
2008 BarrSinoussi,Montagnier HIV;zur Hausen HPV
1997 Prusiner Prions 1997 Prusiner Prions
1989 Bishop,Varmus Retroviraloncogenes
1983 McClintock Transposons Mobilegeneticelements
1975 Baltimore,Dulbecco,Temin Tumour virushostgenetic
interactions
1966 Rous Tumourinducingviruses
1958 Lederberg geneticrecombination;Beadle,Tatum genetic
controlofenzymesynthesis
1952 Waksman StreptomycinagainstTB
1905 Koch CauseofTB
1901 VonBehring Effectivenessofserumagainstdiptheria
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WhereareTheMicrobesTakingUsToday?
BasicMicrobiology
Biotechnology Biochemistry
AppliedMicrobiology
Biotechnology
Systematics
Biochemistry
Virology Genetics
Medical/
Immunology
Agricultural/
Industrial
Aquatic/
Marine
Microbial
Ecology
TaxonomicClassification
Taxonomy:
TheScienceof
namingorganisms
Domains:Archaea /Bacteria/Eukarya
Carolus Linnaeus
(17071778)
Systema Naturae
Kingdoms:Plants/Animals/Fungi
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Greennonsulfur
bacteria
Euryarchaeota
Entamoebae
Slime
molds
Animals
Fungi
BACTERIA ARCHAEA EUKARYA
Mitochondrion
Gram
positive
bacteria
Proteobacteria
Chloroplast
Cyanobacteria
Flavobacteria
Thermotoga
Thermodesulfobacterium
Methanosarcina
Methano
bacterium
Crenarchaetoa
Thermoproteus
Pyrodictium
Thermococcus
Pyrolobus
Marine
Crenarchaeota
Methanopyrus
Thermoplasma
Extreme
halophiles
Fungi
Plants
Ciliates
Flagellates
Trichomonads
Methano
coccus
Figure2.17
Thermodesulfobacterium
Aquifex
LUCA Microsporidia
Diplomonads
(Giardia)
2012 Pearson Education, Inc.
LastUniversal
CommonAncestor
WhatCriteriadoweUsetoClassify
Microorganisms?
Bacteria
Archaea
Methanopyrus
Protista
Amoebaproteus
Bacteria
Viruses
Fungi
Prions
Parasites
Protozoa
Fungi
Lentinula edodes
Virus
Bacteria
Lactobacillussake
Helminths
Arthropods
Lentinula edodes
Retrovirus
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MicrobialClassification
Morphology whatdoesitlooklike?
Physiology howdoesitsustainlife?
Biochemistry what does it produce? Biochemistry whatdoesitproduce?
Molecularanalysis nucleicacidsignature
EvolutionofScientificVoyerism
16651668 1809/1824/1826/18311839 1855 1873
Hookevan LamarckTurpinSchwann Schneider
L h k D t h t B Vi h LeeuwenhoekDutrochet BrownVirchow
FromCorkTo
Chromosomes
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ProkaryotesandEukaryotes
Prokaryote:Beforenucleus
Eukaryote:Truenucleus
WhatstheSame?
Nucleicacids DNA/RNA
C ll b
Nucleobase
Guanine
Cellmembrane
Basicchemicalbuildingblocks:carbohydrates/
proteins/purines andpyrimidines /minerals/fats/
vitamins
Ribosomes
Requireenergyforbasicmetabolism
Highlyregulatedphysiologicsystems
Adenosine
triphosphate
ATP
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...WhatsDifferent?
Eukaryotes Prokaryotes
Nucleus Truenuclearmembraneand
nucleoli
Nonuclearmembraneornucleoli
Chromosome(s) Multiplelinearchromosomes Single,circular,nohistones
withhistones
Celldivision Mitosis Binaryfission
Sexual
reproduction
Meiosis Conjugation
Diameter 10100m 0.22.0m
Membranebound
organelles
Present Absent
organelles
Ribosomes 80S/70Sinorganelles 70Sonly
Cellwall If present,simpleinstructure
nopeptidoglycan
Usuallypresent;complex
peptidoglycan composition
Plasmamembrane Contains receptorscomprised
ofcarbohydrates/sterols
Lackscarbohydrates/sterols
http://www.phschool.com/science/biology_place/biocoach/cells/common.html
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BacteriaandVirusesCompared
Bacteria
Prokaryoticcellularliving
Viruses
Noncellularprotists
organisms
Geneticmaterialismost
oftensinglecircular
chromosomecomprisedof
DNA;freefloatingincell
Produceacute,usually
GeneticmaterialmaybeDNAor
RNA;encasedinproteincoat
Requirehostcellstoreproduce;
mayincorporateintohostDNA
andcauselatentdisease
Mayproduceprotracted(long
lasting) progressive systemic
localizeddiseases
responsivetoantibiotic
therapy
lasting),progressivesystemic
diseases chronichepatitis,
kidneydecline/failure,tumours
Notresponsivetoantibiotic
therapy
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MacrophagesInfectedwith
Mycobacteriumtuberculosis
TCellUnderAttackbyHIV1 y
BacterialForm:Cocci,Bacilli,Spirilla
Lactobacillus
Enterococcus
Treponema
pallidum
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Nutrientand
wastetransport
Respiration
Plasma
membrane
Ribosome
Protein
synthesis
Pili
Attachment
Conjugation
Motility
Flagellum
membrane
Plasmids
Extrachromosomal DNA
Chromosome Chromosome
DNA;Geneticarsenal
CellWall
Shape
Protection
fromosmotic
stress
BacterialCell
Ultrastructure
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HepatitisCVirus(HCV) HumanImmunodeficiencyVirus1(HIV1)
HowdoWeSurvivewithMicrobes?
HowdoTheySurviveUs?
Adaptation
Physiological Adaptation: change in PhysiologicalAdaptation:changein
geneexpressioninresponsetochangesin
theenvironment
GeneticAdaptation:overgrowthofa
m tant to become the predominant t pe in mutant tobecomethepredominanttypein
thepopulation