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127 Cards in this Set
- Front
- Back
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protoza
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unicellular eukaryote, non-photosynthetic chemoheterotrophs
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What are the two stages of protozas?
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cysts- survival
and trophozoites- feeding, reproduction |
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How are protozoa classified?
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By motility
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What is the importance of protozoas?
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They cause devasting diseases- Chagas disease, Entamoeba histolytica, natural insecticides
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Cyanobacteria
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oxygenic photosynthetic bacteria
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Describe the morphology of Cyanobacteria
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unicellular, colonial, filamentous
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What is the importance of Cyanobacteria?
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some fix nitrogen in Heterocysts ( important fertilizer providers) N2> NH4
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Algae
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eukaryotic photoautotrophs, photosynthetic machines
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Phycology
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study of algae
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Describe the morphology of algae
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unicellular, colonial, filamentous
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Why is algae important?
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Agar
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What are the different algaes?
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green algae, red algae, brown algae, diatoms, dinoflagellates
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green algae
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terrestrial plants- precursor
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red algae
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agar source
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brown algae
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kelp, seaweed
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diatoms
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silica cell walls, diatomaceous earth
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dinoflagellates
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plankton, red tides cause neurotoxin
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Transformation
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uptake of naked DNA by a competent cell
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what are some naturally competent organisms in Trans?
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Bacillus, S. pneumoniae, Acinetobacter spp.
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what are some artifically competent organisms in Trans?
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E.coli (important in Biotech)
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In the Trans experiment what were the two organisms used?
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Acinetobacter spp. ADP-1(NC)
and Acinetobacter spp. ADP-6(NC) |
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Acinetobacter spp ADP-1
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Trans experiment ADP-1 strain wild type can grow on POB p-hydroxybenzoate
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Acintobacter spp. ADP-6
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Trans experiment ADP-strain
mutant lacks enzyme protocatechuate oxygenase required to breakdown POB |
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GMM
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trans.- glucose minimal media for control growth
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What media did we use to lyse the organism in the Trans experiment?
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TENS solution to lyse ADP-1
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TENS
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Tris buffer- pH buffer to help preserve DNA
EDTA- chelating agent, to remove cations NaOH- breaks cell walls SDS- detergent to solubilize cell membrane |
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How is ADP-6 transformed?
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ADP-1 lysate cells on GMM
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How can E.coli be artificially competent?
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through chemical treatment
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What enzyme is responsible for the breakdown of POB?
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protocatechuate oxygenase
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What media was used during the transformation experiment?
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GMM- Glucose Minimal Media used for control (ADP-6 grows)
POB- p-hydroxybenzoate media |
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What grows on POB in trans experiment?
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ADP-1 grows and ADP-6 can grow if there is an uptake of lysate cells
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Conjugation
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unidirectional transfer of genetic material requires cell-cell contact (pilus)
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Pilus
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cell-cell contact
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Conj which is the prototroph?
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Pseudomonas putida 503CA
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Conj- which is the auxotroph?
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Pseudomonas putida PAW-15
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Plasmid
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closed-circular, extrachromosal DNA capable of self-replication
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What is the TOL plasmid
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a conjugable plasmid contains genes for catabolism of m-tolulate
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Which strain had the TOL plasmid?
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Pseudomona putida PAW-15
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Which was the donor strain in the conj experiment?
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Pseudomonas putida PAW-15
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Which was the recipient strain in the conj experiment?
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Pseudomonas putida 503CA
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conjugation- Leu-, Tol +
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PAW-15
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conjugation- LEu+ Tol -
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503CA
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what media was used for the conjugation experiment?
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M-tol(m-toluate) none grows
M-tol + leucine PAW-15 grows GMM PAW-15 will not grow |
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How does Pseudomonas putida 503CA synthesize amino acids?
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Through anabolism
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What kind of transfer happened for 503CA to grow from a donor plasmid from PAW-15?
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Vertical
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What are two ways of dna transfer?
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chromosomal and plasmid transfer
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Bacteriophage
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viruses that infect bacteria
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Coliphage
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phage that infect E.coli
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what are the bacteriophage infection pathways?
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lysis- death of host cell
and lysogeny-prophage |
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lytic phages
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produce clear plaques dur to killing of all cells in the zone
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lysogenice phage
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give cloudy plaques because of the growth of lysogens within the zone
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Each plaque produced by a single viral particle or
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PFU/ml plaque forming unit
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# of bacteriophage in a sample is known as a
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titer
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What type of E.coli was used in the sewage filtrate (coliphages)
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E.coli B
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Viruses
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noncellular, obligate intracellular parasites, DNA or RNA (capsid
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Bacteria
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sensitive to antibotices, ATP metabolism, ribosomes,plasma membrane
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virus-bacteria=
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coliphages
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What was the point of the sewage filtrate experiment?
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to observe infection of E.coli and the plaques that formed
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Why did we add bacteria twice in the sewage filtrate?
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to creater larger lawns of cells
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Colisure test
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qualitative test absence/presence of E.coli
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Advantages of colisure test
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easy to use
rapid and accurate results 4hrs |
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What is the marker for fecal contamination in the U.S.?
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E.coli
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What enzyme is present in coliforms?
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B-galactosidase
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What enzyme is present in E.coli?
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B-glucuronidase
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What is the indicator in the colisure test?
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yellow - for all coliforms and E.coli
Red- positive for total coliforms and E.coli |
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What was the uv light used for in the colisure exp?
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E.coli fluoresces under long wave 366nm
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What wave of uv light does E.coli fluoresce under?
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long wave 366nm
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Water analysis membrane filter
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quantitative and reliable method for detection of coliforms
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what is the pore size of filters from the membrane setup?
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0.45um to retain bacteria
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Advantages of water membrane filter
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higher reproducible results, greater sensitivity to larger volumes
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Media used in the membrane filter setup:
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EMB- Eosine methylene blue
selective and diffrential media MacConkey and MUG detects B-glucuronidase (E.coli) |
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What are the inhibitors and indicators for EMB?
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inhibitors- eosin
and indicators methylene blue |
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What is the substrate for EMB?
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lactose
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What are the results for EMB?
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Positive- fermentes pink or dark red
E.coli metallic green sheen Negative- non fermenters- clear |
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What does MacConkey and MUG detect?
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B-glucuronidase (E.coli)
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What are inhibitors for MacConkey and MUG?
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crystal violet and bile
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Indicators for MacConkey and MUG
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pH indicator neutral red for lactose fermentation
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What is the wavelength used to intrepret the results of MacConkey and MUG?
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366 nm
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Glucose and Lactose
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determine fermentation
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Indicators for glucose and lactose
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positive- acid yellow color
gas- bubble negative- red color stays |
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Enterotube
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rapid identification of Enterobacteriacase
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What are the two requirements for enterotubes?
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gram - rods
and oxidase - |
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MPN
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most probable number of organisms
estimate |
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Media used for MPN
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BGBLB- diffrential media, lactose as fermentable sugar
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BGBLB
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Brillant green bile lactose broth
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Inhibitors for BGBLB
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brillant green and bile for non-coliforms
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BGBLB results
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pos- yellow lactose fermentation
neg- green |
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Commensalism
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one benefits and other not harmed or helped +/0
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organisms used in Microbial relationship experiments
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Clostridium sporogenes- gram +rods (benefits)
Staphylococcus spp.- gram + cocci (not affected) |
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Antagonism
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one benefits and other is harmed +/-
(production of substance (antibiotics) |
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Antibiotics
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natural chemicals produced by soil organisms
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Antagonism exp organisms used-
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Streptomyces griseus gram + actinmycetes branched and filamentous prokaryotes
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Streptomyces griseus
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round chalky colonies
give unique earthy smell- geosmin |
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Geosmin
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organism that gives Streptomyces griseus earthy smell
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Why did we use water agar in the antagonism exp?
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so the organism would grow on top
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Bacterial count in food and milk importance
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milk good growth media for microorganisms, # indicates freshness of milk
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bacterial count in meats=
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relationship to meat spoilage
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dilutions
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to get a countable # of organisms (30-300)
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Epidemiology
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study when and where diseases occur and how they're transmitted to the human population and who is spreading the disease
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Pandemic
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epidemic disease- worldwide
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Endemic
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constantly present in the population
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Sporadic
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Occasionally
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Communicable diseases
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transmissible to other persons through direct contact and indirect contact
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pathogen
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Serratia marcescens red
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normal microbiota
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Micrococcus luteus yellow
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SIM
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hydrogen sulfide, idole, motility
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Sulfide
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H2S production, reduction of sulfur
ferrous salts reacts with H2s pos- black precipitate |
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Indole
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differentiate btwn enteric bacteria which are able to cleave indole from Tryptophanase enzyme
Pos- red ring (rosindole dye) |
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What reagant did we add to Indol?
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Kovac's reagant
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What enzyme is sometimes cleaved in Indole?
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Tryptophanase enzyme
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Motility
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ability of an organism to move from inoculation site to cause turbidity
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Urea
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determine the presence of urease enzyme
pH 6.8 orange neg pos ph 8.1 ammonia bright pink |
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MR
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methyl red ability of organisms to ferment glucose
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indcator for MR
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methyl red
pos red pH 4.5 neg yellow pH 6,7 |
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VP
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Voges- Proskauer
producing neutral products acetoin detection |
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what solution did we add to Voges- Proskauer?
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Barrit's solution A and B
alpha naphthol and KOH |
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Indicators of voges proskaure
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positive- red color on top of the broth
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citrate
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using as sole carbon source for media
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What must an organism have to take up citrate?
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transport system or Citrate permase
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what is the indicator for ctirate
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positive Bromothymol blue
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Gram stain
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gram + purp
gram - pink crystal violet, iodine, decolorizer, safranin |
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Oxidase test
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detects for cytochrome oxidase enyzme complex in etc
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indicators for oxidase test
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pos purp
neg yellow tetramethyl-p-phenylenediamine |
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Starch hydrolysis
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starch used as carbon source to determine the presence of amylases
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starch
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glucose, maltose, dextrin
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indicator for starch hydrolysis
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iodine ( test for the disappearance of starch)
pos- clear halo around colony |
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catalse test
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hydrogen peroxide to organism to test for catalse activity
pos bubbling |
