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Microbialbiotechnologyinthelaboratoryandinpractce
3.
Inoculatefromappropriatedilutionsofthesoilextract
(Figure1.2.5)0.1mlperplateinthefollowingmedia:
agar,ZT,agarwithanthraceneorphenanthrene(0.1g/l),
mediumwithmilk,withTween80,withstarch.
Inoculateduplicatesfromeachdilution.Incubatethe
plates48hfor7daysat28°C.
4.
Performmacroscopicobservationofthecultures,count
thenumberofmicroorganismsin1gofsoil.
5.Isolationofsinglemicrobialcolonies:
a)
producingproteases(lighteningaroundthecolonieson
plateswithmilkcontainingmedium);singlecolonies
shouldbetransferredtoplateswithmilkandastreak
cultureshouldbecarriedout;
b)
Actinomycetes;singlecoloniesshouldbetransferred
tomaltoseplatesandinoculatedusingtheloop.
After7days,examinetheantagonisticpropertiesof
theselectedsoilactinomycetesinrelationtothetest
bacteria:Staphylococcusaureus,Escherichiacoli,
Pseudomonasaeruginosa;
c)
microorganismspotentiallycapableofdecomposing
petroleum-basedcompounds(anthraceneand
phenanthrene);singlecoloniesshouldbetransferred
toauniversalmedium.
Additionally,amicroscopicpreparationofthe
isolatedmicroorganismshouldbemade.
Isolatesofmicroorganismscapableofproducing
proteolyticenzymesthatproduceasubstancethatis
antagonistictothetestedbacteriashouldbesecured
forfurtheranalyses.
6.
Evaluationofproteolyticenzymeactivityof
microorganismsisolatedfromthesoil(seeSection4.1.3).
7.
Evaluationoftheactivityofsubstancesthatare
antagonistictothebacteriabeingtested,producedby
microorganismsisolatedfromthesoil(seeSection4.1.5).
8.
Evaluationoftheabilityofisolatedstrains(bacteriaand
fungi)todegradepetroleum-basedcompounds:
a)
streakingofselectedcoloniesfromplatesonuniversal
(bacteria)andZTslants(fungi);
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