1
Q
Objective lens
A
- 4 different objective lens
- 4x
- 10x
- 40x
- 100x (oil immersion objective)
2
Q
total magnification
A
- magnification by the objective lens x magnification by the ocular lens (always 10x)
- ex. (4)(10)= 40x
3
Q
condenser
A
- gathers light
- conducts light into the slide
4
Q
x and y mechanical stage knobs
A
- use this to move the cells into field
- move the bright light into field by using these
5
Q
ocular lens
A
- stays constant
- 10x
6
Q
oil immersion
A
- before moving to 100x place an oil immersion drop onto the slide
- you will be a bright circle before
7
Q
coarse focus
A
-never use 40x and above
8
Q
10x
A
use to locate the object
9
Q
cocci/coccus
A
spherical
10
Q
bacilli/bacillus
A
rod shaped
11
Q
sprillum
A
loose spiral
12
Q
spirochete
A
tight spiral
13
Q
bacteria
A
- exists in 3 shapes- spherical, rod, spiral
- no nucleus
- prokaryote
- *shape and arrangement are the main characteristics of bacteria
14
Q
arrangements of bacteria
A
- chains -> streptococci, streptobacilli
- groups of two -> diplococci, diplobacilli
- staphylococci -> cluster
15
Q
media
A
- aka nutrients
- media is used to grow microbes
- have nutrients for bacteria to grow
- includes:
- glucose- energy
- peptones- breakdown products of proteins -> good source of amino acids
- yeast extract has vitamins
16
Q
liquid medium
A
- nutrient broth
- use when you want to make a large amount of culture
17
Q
solid medium
A
- nutrient agar
- jelly like
- agar comes from red algae and it is a polysaccharide
- agar is a solidifying agent
- nutrient agar slant- slanted surface is used to grow culture
- solid medium has glucose, peptones, yeast in addition to agar
18
Q
making nutrient agar
A
- granules of agar are added to nutrient broth
- boil the mixture until the agar goes into the solution
- cool solution to 40C -> becomes a solid
19
Q
agar: good solidifying agent
A
- stays solid in the incubator
- microbes cannot metabolize agar (breakdown)
- allows us to study colony morphology
- solid allows us to isolate a specific species of microbe
20
Q
sterilizing media
A
- use autoclave
- media must be free of microbes
- autoclave must have temperature of 121C, pressure of 15psi, for 15 minutes
- heat labile (sensitive) solutions are sterilized by using filtration
21
Q
filtration
A
- sterilization
- used for solutions that are sensitive to heat (enzymes)
- microbes that are in the solution stay on the surface of the filter
- separates the microbes from the solution
- does not kill microbes
- .22um
22
Q
platyheminthes
A
flatworms
- two types: trematodes (flukes) & cestodes (tapeworms)
- flat body
- hermaphroditic- male and female reproductive organs
- both testes and ovaries
- complex life cycle
- they need a intermediate host and definitive host
23
Q
nematodes
A
roundworms
24
Q
trematodes
A
- flukes
- flat leaf body
- a type of platyheminthes
25
intermediate host
- larvae of the worm (immature worm)
| - asexual reproduction takes place
26
definitive host
- adult worm (mature worm)
| - sexual reproduction takes place
27
Clonorchis sinensis
- trematode (fluke)
- chinese liver fluke
- shaped like a leaf
- lives in the liver of infested people
- *shaped like flower vase
- *operculum- lid like structure on one side and smooth on the other
28
cestodes
- tapeworms
- flatworms (platyheminthes)
- scolex- head
- segmented body
- segment- proglottids
- hermaphroditic
- each proglottid has testes and ovaries
- can be very long
29
taenia saginta
- beef tapeworm
- adult- human intestinal tract
- eggs are released in the feces
- ingested by cattle
- larvae come out in the intestinal tract
- cysts in the muscle
- cyst (sleeping, resting larva)
- undercooked, contaminated meat ingested by humans
- scolex comes out of the cyst
- attaches to the intestinal wall
- makes proglottids (longer)
30
cyst
- larvae sleeps in the cyst
| - dormant
31
tapeworm characteristics of ova
- granule center
| - striated ring
32
nematodes
- roundworms
- dioecious
- male and female worms
33
entirobius vermicularis
- pinworm
- nematode
- intestinal tract of humans
- female worm comes out of anus
- releases its eggs
- eggs- clothing, bedding, airborne
- ingested by another host
- eggs reach intestinal tract
- larvae come out
- grows into an adult worm
- intense itching in the perianal area
34
nematode, pinworm, entirobius vermincularis characteristics of ova
- elongated
- granule center
- oval shape
- one side is flat while the other is more curved
35
ascaris lumbricoides
- roundworm
- adult- intestinal tracts
- eggs released in the feces
- ingested in contaminated food and water
- larvae come out in the intestinal tract
- grow into adult worms
- eat semidigested food
- blockage (obstruction) of the intestinal tract
36
ascaris lumbricoides charactertistics of ova
- irregular surface (bumpy)
| - granule center
37
necator americanus
- hookworm
- adult- intestinal tract
- eggs- feces (eggs are not infected- no problem)
- larvae come out in the soil
- when people step on them, they penetrate the skin
- migrate to the intestinal tract
- attach to the intestinal wall
- feed on blood
- grow into adult worms
- anemia
- prevented by wearing shoes
- larvae is infected not eggs
38
necator americanus characteristics of ova
- lumps of granules
| - sac of potatoes
39
trichuris trichiura
- whipworm
- adult- intestine
- eggs- feces
- contaminated food or water
- larvae come out in the intestinal tract
- bloody diarrhea
40
trichuris trichiura characteristics of ova
- dark outline of structure
- football shape
- granule center
41
trichinella spiralis
- nematode, roundworm
- transmitted in contaminated, undercooked
- encysted pork (get infested depending on their environment (trash))
- larvae come out of the cyst in the intestinal tract
- grow into adult worms
- female releases larvae
- from cysts in the muscle
- diaphragm muscle (affects breathing)
- worm doesnt release eggs so we look for the cyst in the muscle to diagnose
- avoid by cooking meat well
42
protozoa
- eukaryotic
- unicellular
- two stages:
- trophozoite stage
- cyst stage
43
trophozoite stage
- metabolically active
- chemical rxns
- uses nutrients
- motile
- most cases this stage is found inside the host
44
cyst stage
- dormant stage
- resting
- no chemical rxn
- most cases found outside the host
45
entamoeba histolytica
- pathogenic protozoa
- cysts are released in the feces of infected
- cysts are smaller and have 4 nucleus
- transmitted in contaminated food or water
- when the cysts are ingested, they reach intestinal tract
- become trophozoites (larger and has one nucleus)
- they feed on cells lining the intestinal tract
- ingest RBC
- cause dysentery (blood diarhea)
46
trichomonas vaginalis
- pathogenic protozoa
- transmitted by sexual contact
- itching in the area
- single nucleus
- bunch of flagella on one end of the cell
- only has trophozoite stage (no cyst stage)
47
giardia lamblia
- pathogenic protozoa
- cysts are released in the feces
- cysts have oval shape, flagella are inside the cell, and at least two nuclei
- contaminated food and water
- cysts transform into trophozoites (have flagella)
- attach themselves to the intestinal wall and absorb nutrients
- diarrhea and weight loss
- diagnosed based on presence of cysts
48
trypanosoma gamiense
- pathogenic protozoa
- causes african sleeping sickness
- transmitted by tsetse fly
- injected into the bloodstream
- toxins damage certain parts of the nervous system
- causes confusion, drowsiness, coma, death
- there is no cyst stage (exists of trophozoites)
- long slender body
- long single flagellum
- wavy membrane
- one nucleus
49
trypanosoma cruzi
- pathogenic protozoa
- causes chagas disease
- transmitted by kissing bug
- protozoa gets into the blood stream
- toxins damage certain parts of the nervous sytstem
- causes mega colon, mega esophagus (enlargement) -> constipation
- has a flagellum and nucleus
- dark blue dot on one end of the cell is the mitochondrial DNA
- smaller and thinner than trypanosoma gamiense
50
kinetoplast
mitochondrial DNA
51
balantidium coli
- pathogen protozoa
- cysts are released in the feces
- contaminated water or food
- become trophozoites in the intestinal tract
- can cause dysentery (bloody diarrhea)
- cyst stage- macronucleus
52
plasmodium falciparum
- pathogen protozoa
- causes malaria
- transmitted by mosquito
- forms a ring stage in the RBC
- diagnosis is based on the presence of the ring stage
53
aseptic procedure
- necessary bc mircrobes are ubiquitous
- used to make pure cultures
- prevents us from contaminating ourselves with microbes
54
pure culture
- only one species of microbe
| - needed to study specific microbes
55
steps of aseptic procedure for serratia marcescens
- we need a sterile nutrient agar slant, culture of serratia marcescens (red), bacti-cinerator, inoculating loop, labels, gloves
1. sterilize inoculating loop in the bacti-cinerator for 5s and cool for 5s
2. hold the tubes in nondominant hand and decap with the other
3. transfer serratia marcescens into steril nutrient agar slant
4. cap the tubes
5. sterilize inoculating loop in the bacti-cinerator
6. incubate at 30C for 24-48 hours
7. pure culture is made!
56
bacti-cinerator
- takes 10 mins to heat to orange
- sterilizes the inoculating loop
- wait 5 secs
57
aseptic procedure for pure broth culture of E. coli
1. sterilize inoculating loop in the bacti-cinerator for 5s and cool for 5s
2. hold the tubes in nondominant hand and decap with the other
3. dip loop into E. coli and then dip into sterile broth -> shake
4. cap the tubes
5. sterilize inoculating loop in the bacti-cinerator
6. incubate at 30C for 24-48 hours
7. cloudy pure culture is made!
58
Escherichia coli culture
- E. coli
- broth is cloudy
- teeming with bacteria
59
streaking for isolation
- isolates specific species of bacteria from a sample that has many different species of microbes
- done to study colony morphology
- can be used to see if a culture is pure or contaminated
- gradually dilution culture
60
petri plate- nutrient agar plate
- smaller plate has the agar
- larger plate is a lid
- always label the bottom (small plate)
- plates are incubated upside down
- smaller plate face up
61
streaking for isolation procedure
1. divide the bottom of the plate into 3 sections
2. label
3. sterilize loop
4. hold culture
5. remove cap
6. insert loop
7. cap
6. transfer the culture to the section 1 of plate in a zig zag manner
7. cap
8. sterilize
9. zig zag from 1 to 2 about four times and then spread throughout 2
10. cap
11. sterilize
12. zig zag from 2 to 3 about 4 times and then zig zag throughout section 3
13. cap
14. sterilze
15. incubate at 30C for 24-48 hours
16. isolated colony in section 3
62
colony morphology
- shape, color, texture
| - try to identify genus and species
63
confluent growth
-section 1 has growth all over the medium
64
isolated colony
section 3
- separate and identical
- we know the culture isnt contaminated
65
colonies close together
- section 2
- colonies are touching
- not isolated
66
wet mount procedure
- take a slide
- place a drop of sample in the center
- place a coverslip over sample
- observe under 10x and 40x objective lens
67
hay infusion wet mount
- taking a few strands of hay
- soak the hay in water for a couple of weeks
- water becomes brown liquid
- teeming with microbes
- bacilli
- cocci
- alive
68
pond water wet mount
- algae
- green
- sometimes worms, diatoms
69
amoeba wet mount
- pseudopod (plasma membrane folds outward, used to crawl) are apparent
- granules- floating and moving
- cytoplasmic streaming- flowing of granules (moving)
- ameba is crawling
70
euglena wet mount
- flagellum
| - green- chloroplasts
71
paramecium wet mount
- protozoa
- cilia
- cell is elongated
- cytosome- mouthlike structure, groove
72
borrelia burgorferi
-lymes disease
73
things that affect gram staining
- prolonged discoloration - G+ -> G-
- old culture- G+ -> G-
- too much heat fixing- G+ -> G-
- thick smear- G- -> G+
74
colony
- millions of identical cells clump
- come from the same single cell
- not touch
Microbiology
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