P1 Bacterial Cultivation (LECTURE) Flashcards
prelims
1
Q
Refers to the increase in the number of bacteria rather than in size
- bacteria grow in number and do not grow in size
- This growth is affected by various factors such as optimum growth requirements, dynamics of growth, including the use of a medium that can be artificially prepared in the laboratory
A
Bacterial Growth
2
Q
Bacterial Growth Requirements
A
- Nutrition
- Environmental
3
Q
NUTRITIONAL REQUIREMENTS:
(C.ES.ES.N.W.ME.S.A/SGR.GR.TR.PHR)
A
- Carbon
- Energy Source
- Electron Source
- Nitrogen
- Water/Moisture/Humidity
- Mineral Elements
- Salt
- Additional/ Special Growth Requirements
- Gaseous Requirement
- Temperature Requirement
- pH Requirement
4
Q
Needed for the synthesis of cellular components
A
Carbon
5
Q
Carbon is at least ____ of weight
A
50%
6
Q
Carbon Dioxide from the air
A
Autotroph
7
Q
Organic compounds in the culture media (glucose)
A
Heterotroph
8
Q
Energy source is ______ of weight?
A
4%
9
Q
Light?
A
Phototroph
10
Q
Chemical Energy?
A
Chemotroph
11
Q
Electron Source
(I.O.N.F) “INOFF”
A
- Inorganic molecule (Fe2+)
- Organic molecule
- NADH reduced from NAD
- FADH2 reduced from FAD
- Inorganic molecule - litotroph
- Organic molecule - organotroph
12
Q
Needed for the synthesis of proteins
A
Nitrogen
- Free nitrogen from the air
- Nitrogenous compounds in the culture media (e.g., peptone, yeast, beef extract)
13
Q
Nitrogen is (___) of weight?
A
14%
14
Q
Water/ Moisture/ Humidity is (_________) of bacteria
A
70%
15
Q
Needed as co-factors in various metabolic process of the bacteria
A
Mineral Elements
16
Q
Bacteria can tolerate salt concentration below (____)?
A
6%
17
Q
certain bacteria that can survive high salt concentration environment
A
Halophilic bacteria or Halophiles (salt loving)
18
Q
Bacterias that are Halophilic or Halophiles (salt loving):
(S.E.V.B)
A
- Staphylococcus spp.
- Enterococcus spp.
- Vibrio spp. (except: Vibrio cholerae & Vibrio mimicus)
- Bacillus spp.
19
Q
Bacteria that are very difficult to grow
A
Fastidious
- requires special or additional requirements to grow in culture media
20
Q
Example of blood loving organism?
A
Haemophilus spp.
21
Q
Requires both X & V factor
A
Haemophilus spp.
22
Q
Additional/ Special Growth Requirements
Culture media:
* intact RBC
* x factor only
A
Blood Agar Plate
23
Q
Culture media: Blood Agar Plate
1st priority:
A
5% defribinated sheep’s blood
24
Q
Culture media: Blood Agar Plate
2nd priority:
A
horse blood
25
# Culture media: Blood Agar Plate
3rd priority:
Rabbit's blood
26
# Culture media: Blood Agar Plate
Least priority:
Human blood
| (must be type O – no presence of antigens)
27
Since blood agar plate lacks V factor, use (____)?
Chocolate Agar Plate
28
contains NAD – v factor
| * lysed RBC (heat)
* has v and x factor
Chocolate Agar Plate
29
hydrolyzes the NAD
NADase
30
hydrolyzed NAD means?
| NAD = Nicotinamide Adenine Dinucleotide
No V factor
31
Degradation product of Hgb (hemoglobin)
X Factor (Hemin/ Hematin)
32
# Gaseous Requirement
Bacteria that grow, live, and survive in the presence of oxygen
Aerobe
33
# Gaseous Requirement
Absolutely requires oxygen to grown, live, and survive
Strict/Obligate Aerobe
34
# Gaseous Requirement
Example of Strict/Obligate Aerobe bacteria?
## Footnote
(M.M.P.N.B.F.B.L) - spp.
* *Micrococcus spp.*
* *Mycobacterium spp.*
* *Pseudomonas spp.*
* *Neisseria spp.*
* *Brucella spp.*
* *Francisella spp.*
* *Bordetella spp.*
* *Leptospira spp.*
35
# Gaseous Requirement
Bacteria that have the ability to grow, line and survive in small concentration of oxygen environment
Facultative Anaerobe
36
# Gaseous Requirement
Example of Facultative Anaerobe bacteria?
## Footnote
(S.S.FE)
* *Staphylococcus spp.*
* *Streptococcus spp.*
* *Family Enterobacteriaceae*
37
# Gaseous Requirement
Bacteria that prefers small concentration of oxygen environment
* approximately (_____)?
Microaerophilic
* 2%-10%
38
# Gaseous Requirement
Example of Microaerophilic bacteria?
## Footnote
(C.H.A.S) - spp.
* *Campylobacter spp.*
* *Helicobacter spp.*
* *Arcobacter spp.*
* and some *Streptococcus spp.*
39
# Gaseous Requirement
Bacteria that grow, live and survive in the absence of oxygen
Anaerobe
40
# Gaseous Requirement
Absolutely do not requires oxygen to grown, live, and survive
Strict/ Obligate Anaerobe
41
# Gaseous Requirement
Example of Strict/Obligate Anaerobe bacteria?
## Footnote
(C/CN.B.FB.PS.PR) - spp.
* many *Clostridium spp.* (*Clostridium noyvi*)
* Most *Bacteroides spp.*
* *Fusobacterium spp.*
* *Peptostreptococcus spp.*
* *Porphyromonas spp.*
42
# Gaseous Requirement
Bacteria that do not require oxygen but may ***tolerate*** or ***withstand*** limited exposure to oxygen
Aerotolerant Anaerobe
43
# Gaseous Requirement
Example of Aerotolerant Anaerobe bacteria?
## Footnote
(C [CP,BF])
(PB & LB)
* Some *Clostridium spp.* (*Clostridium perfringens*, *Bacteroides fragilis*)
* Most strains of *Proprionibacterium* & *Lactobacillus*
44
# Gaseous Requirement
Bacteria that requires 5%-10% CO2 to grow
Capnophilic
45
# **Gaseous Requirement:** Capnophilic
**N.H.A.CB.E.K.SP GROUP**
* *Neisseria spp.*
* *Haemophilus spp.*
* *Aggregatibacter spp.*
* *Cardiobacterium spp.*
* *Eikenella spp.*
* *Kingella spp.*
* *Streptococcuse pneumoniae*
46
Most pathogenic bacteria would grow at temperature between 35-37C
## Footnote
* hence incubator in the laboratory is usually set and maintained within this temperature range for routine isolation of pathogens
Temperature Requirement
47
# **Temperature Requirement**
* Mesophilic
* Psychrophilic/Cryophilic
* Thermophilic
* Hyperthermophilic/ Extremely Thermophilic:
* Eurithermophilic:
* Sternothermophilic:
* 20-40C
* 0-20C
* 50-60C
* 80-110C
* wide range
* narrow range
48
most pathogenic bacteria can grow in a neutral or slightly alkaline environment
## Footnote
hence most culture media used in routine isolation of pathogens is adjusted to this (____)
pH Requirement
## Footnote
(pH range)
49
pH requirement (___)
pH 7.0 – 7.5
50
# pH Requirement
Acid loving bacteria
Acidophilic
51
# pH Requirement
Example of acid loving bacteria?
## Footnote
(LB.A)
*Lactobacillus acidophilus*
| normal flora of vagina
52
# pH Requirement
Alkali loving bacteria
Alkaliphilic
53
# pH Requirement
Example of alkali loving bacteria?
## Footnote
(GV)
Gardnerella vaginalis
54
# pH Requirement
Osmophilic bacteria
High Osmotic Pressure
55
# pH Requirement
Example of osmophilic bacteria
## Footnote
(AB spp.)
*Archaebacteria spp.*
56
# Bacterial Growth Phase
* Generation time
* Fast Growing Bacteria
* Slow Growing Bacteria
* Time to replicate & Refers to the stages of bacterial growth
* 20 minutes
* 24 hours
57
4 Bacterial Growth Phase:
1. Lap Phase
2. Log/Logarithmic/Exponential Phase
3. Maximum stationary/Plateau phase
4. Decline/Death phase
58
# Bacterial Growth phase:
Bacteria are still adjusting to the new environment hence there is no cell division that occurs in this phase
## Footnote
they are activity synthesizing DNA & proteins that are necessary for cell division
Lag Phase
59
# Bacterial Growth phase
The number of generation per hour
growth rate constant
60
# Bacterial Growth Phase
Phase where there is a sudden increase of bacteria because of rapid generation or doubling time.
## Footnote
* also the phase where bacteria are most metabolically active hence most susceptible to the action of antimicrobial agents
Log/Logarithmic/Exponential phase
61
# Bacterial Growth phase
After essential nutrients are depleted, toxic products accumulate or oxygen becomes limiting, the rate of cell division equates the rate of cell death
Maximum stationary/Plateau phase
62
# Bacterial Growth Phase
Due to unfavorable environment for growth, bacterial cell division decreases while cell death becomes more accelerated
Decline/Death phase
63
# Bacterial Growth phase: answer whether it has C.D and Increase in Number
Lag phase
| C.D = Cell Division
## Footnote
(YES OR NO)
* **Cell Division:** NO
* **Increase in Number:** NO
64
# Bacterial Growth phase: answer whether it has C.D and Increase in Number
Log/Logarithmic/Exponential phase
| C.D = Cell Division
## Footnote
(YES OR NO)
* **Cell Division:** YES
* **Increase in Number:** YES
65
# Bacterial Growth phase: answer whether it has C.D and Increase in Number
Maximum stationary/Plateau phase
| C.D = Cell Division
## Footnote
(YES OR NO)
* **Cell Division:** YES
* **Increase in Number:** NO
66
# Bacterial Growth phase: answer whether it has C.D and Increase in Number
Decline/Death phase
| C.D = Cell Division
## Footnote
(YES OR NO)
* **Cell Division:** YES
* **Increase in Number:** NO
67
artificial preparation in the laboratory which contains basic foundation of nutrients and a solidifying agent (if needed) to support the growth of microorganisms
## Footnote
* Additional substances may be added to enrich the media for growth of microorganisms that are very difficult to grow (FASTIDIOUS)
CULTURE MEDIA
68
# **TERMINOLOGIES:** Culture Media
**CULTURE**
* Noun:
* Verb:
* growth of microorganisms
* to growth/ to cultivate microorganism
69
# **TERMINOLOGIES:** Culture Media
Introducing the microorganism to the culture media
Inoculate/Plant/Cultivate
70
# **TERMINOLOGIES:** Culture Media
Transfer of microorganisms from one culture media to another
Transplant/ Subculture
71
# **Classification of Culture Media**
Composed of known and exact amounts of pure chemical substances
| * *according to Composition*
## Footnote
(S/CD)
Synthetic/Chemically Defined
72
# **Classification of Culture Media**
Composed of complex materials that are rich in vitamins and nutrients that are not usually represented by a chemical formula such as peptones, beef or yeast extract, plant extracts etc.
| * *according to Composition*
## Footnote
(N-S, NCD, C)
Non-Synthetic/Non-Chemically Defined/Complex
73
# **Classification of Culture Media**
Live cells harvested from organs of humans and animals that supports the growth of obligate intracellular organisms that cannot grow in artificially prepared culture media
| * *according to Composition*
## Footnote
(TCM)
Tissue Culture Media
74
Tissue Culture Media used in Microbiology Laboratory:
## Footnote
(VCL, MCCCL, CE, AC, HCL, H2CL)
* Vero Cell Line
* Mc Coy Cell Line
* Chicken Embryo
* A549 Cells
* HELA Cell Line
* Hep-2 Cell Line
75
# Tissue Culture Media used in Microbiology Laboratory
Kidney cells of an African Green Monkey
| source
Vero Cell Line
76
# Tissue Culture Media used in Microbiology Laboratory
Mouse cell line
| source
Mc Coy Cell Line
77
# Tissue Culture Media used in Microbiology Laboratory
Fertilized chicken egg
| source
Chicken Embryo
78
# Tissue Culture Media used in Microbiology Laboratory
Human lung carcinoma
| source
A549 Cells
79
# Tissue Culture Media used in Microbiology Laboratory
Human cervical carcinoma
| source
HELA Cell Line
80
# Tissue Culture Media used in Microbiology Laboratory
Human epithelial cells of larynx carcinoma
| source
Hep-2 Cell Line
81
# Classification of Culture Media according to Physical State/Consistency
a culture medium that doesn’t contain a solidifying agent
Liquid
82
# Classification of Culture Media according to Physical State/Consistency
A culture medium that contains 0.5% - 1% agar
Semi-solid
83
# Classification of Culture Media according to Physical State/Consistency
Used for observation of hydrogen sulfide gas production, **indole** production and motility
| under semi-solid
## Footnote
(SIMM)
Sulfide Indole Motility (SIM) Medium
84
# Classification of Culture Media according to Physical State/Consistency
A culture medium which contains 1.5% - 3%T (2%-3%) agar
Solid
85
# Classification of Culture Media according to Manner of:
Usually contained in a container that can be made of glass (pyrex) or disposable plastic (Petridish)
| Dispensing/Formation
Plated
86
# Classification of Culture Media according to Manner of:
Usually, container in glass tubes such as Wassermann tubes with different volume capacity (3mL, 5mL, 10mL) or in a tube with a **flat bottom** and **a screw cap**
| Dispensing/Formation
Tubed
87
# Classification of Culture Media according to Manner of:
Culture media contained in a glass bottle that is usually used for blood culture
| Dispensing/Formation
Bottled
88
# **Classification of Culture Media according to Function/ Use**
Contains basic nutritional requirements to support the growth of non-fastidious microorganisms
## Footnote
This is also used a base medium in the preparation of other culture media
General Purpose/Primary/Basic/Basal/ Supportive/General Isolation Culture Media
89
# **Classification of Culture Media according to Function/ Use**
Contains the basic nutritional requirement to support the growth of non-fastidious microorganisms with additives, **enriching** substances, or supplements to support the growth of fastidious microorganisms
## Footnote
(e.g. Blood Agar & Chocolate Agar Plate)
Enriched Culture Media
90
# **Classification of Culture Media according to Function/ Use**
A primary media used to support or favor the selective growth of pathogens in a specimen, such as stool or sputum, where the number of normal flora outnumber the pathogens
| * Use commonly stool specimen
## Footnote
(EB)
Enrichment Broth
91
Examples of Enrichment Broth:
## Footnote
(APW, SFB, THIOB, GNB,T-HB)
* Alkaline Peptone Water (APW)
* Selenite F Broth
* Thioglycolate Broth (THIO)
* GN Broth (Gram Negative Broth)
* Todd-Hewitt Broth
92
Used to selectively favor the growth of *Vibrio* while inhibiting all other normal intestinal flora due its high pH
Alkaline Peptone Water (APW)
93
Used to selectively favor the growth of *Salmonella* while inhibiting all other normal intestinal flora
Selenite F Broth
94
An enrichment broth for anaerobic bacteria but should be used solely in the isolation of anaerobic bacteria since it can also grow aerobes and facultative anaerobes
## Footnote
(THIOB)
Thioglycolate Broth (THIO)
95
Used to selectively favor the growth of Salmonella and Shigella while inhibiting all other normal intestinal flora
GN Broth (Gram Negative Broth)
96
A liquid enrichment recommended for the production of Streptococcal haemolysin and the cultivation of Streptococci prior to serological grouping
Todd-Hewitt Broth
97
A primary isolation culture media which maintains the viability of bacteria allowing rapid multiplication if there is an anticipated delay in bringing the specimen collected bedside or remotely to the laboratory
## Footnote
(TCM)
Transport Culture Media (TCM)
98
Favors the growth of the organism of interest using inhibitors added in the culture media
## Footnote
(SCM)
Selective Culture Media
99
Contains indicators which changes in color as a result of a product produced be a chemical reaction in the components of the media such as glucose
## Footnote
(DCM)
Differential Culture Media
100
# **NOTE**:
it is important to remember and understand that **not all selective culture media** are ***differential*** but **all differential culture media** are ***selective***
101
# **INHIBITORS IN CULTURE MEDIA**
* **Dyes:**
* **Chemicals:**
* **Antibiotics:**
| Inhibitors for Gram Positive Bacteria
## Footnote
(CV,E,MB,BG,ETC)
(BMS,BS[SD],TS,C,ETC.)
(V)
* Crystal violet, eosin, methylene blue, brilliant green, etc.
* Bismuth sulfite, bile salts (sodium desoxycholate), thiosulfate, citrate, etc.
* Vancomycin
102
# **INHIBITORS IN CULTURE MEDIA**
* **Dyes:**
* **Chemicals:**
* **Antibiotics:**
| Inhibitors for Gram Negative Bacteria
## Footnote
(BF & T [BA])
* Basic fuchsin and
* thionine for *Brucella abortus*
103
# **INHIBITORS IN CULTURE MEDIA**
* **Antibiotics:**
| Inhibitors for Fungi
## Footnote
(N.A.AB)
* Nystatin
* Anisomycin
* Ampothericin B.
104
# **INHIBITORS IN CULTURE MEDIA**
* **Indicators in Culture Media**:
* Dyes and chemical substances such as pH indicators
105
# **Eosin Methylene Blue (EMB) Agar**
* **Original Color:**
* **Selective for:**
* **Inhibitors:**
* **Differential Indicators:**
| (DV)
(GNEB)
(E&MB)
(E&MB)
## Footnote
MOST COMMONLY AND ROUTINELY USED SELECTIVE & DIFFERENTIAL CULTURE MEDIA IN THE LABORATORY
* Dark violet
* Gram negative enteric bacilli
* Eosin and Methylene Blue
* Eosin and Methylene Blue
106
**RLF:** Rapid Lactose Fermenter
| Number of hrs of incubation
able to ferment lactose for **18-24 hours** incubation
107
**LLF:** Late Lactose Fermenter
| number of hrs of incubation
able to ferment lactose for **36-72 hours** incubation
108
# **RLF**
medium sized dark violet colonies with greenish metallic sheen
| (EC)
## Footnote
Differentiation of Family Enterobacteriaceae based on lactose fermentation
**Escherichia**
(*E. coli*)
109
# **RLF**
Appears medium sized dark violet colonies with/without dark center
| (EB)
## Footnote
Differentiation of Family Enterobacteriaceae based on lactose fermentation
*Enterobacter*
110
# **RLF**
Large sized mucoid dark violet colonies with or without dark center
| (KS)
## Footnote
Differentiation of Family Enterobacteriaceae based on lactose fermentation
*Klebsiella*
111
LLF bacteria:
## Footnote
(HSCB,SA,SS,YEC)
* *Hafnia, Serratia, Citrobacter*
* *Salmonella arizonae*
* *Shigella sonnei*
* *Yersinia enterocolitica*
112
NLF (appear colorless colonies) bacteria:
## Footnote
(PPME)
* **All** *Salmonella* **except** *S. arizonae*
* **All** *Shigella* **except** *S. sonnei*
* **All** *Yersinia* **except** *enterocolitica*
* *Proteus, Providencia, Morganella, Edwardsiella*
113
# **Mc Conkey (MAC) Agar**
* **Original Color:**
* **Selective for:**
* **Inhibitors:**
* **Differential Indicators:**
## Footnote
(LP)
(GNEB)
(CV, BS, C)
(NR)
* Light pink
* Gram negative enteric bacilli
* Crystal violet, bile salts, Citrate
* Neutral Red
114
# **Salmonella-Shigella (SSA) Agar**
* **Orginal color:**
* **Selective**
***For:***
***Inhibitors:***
* **Differential:**
***Indicators: pH indicators:***
***H2S indicator:***
***Sulfur Source:***
## Footnote
(LO)
(S & SG spp.)
(BG, BS, C)
(NR)
(FAC)
(ST)
* **Orginal color:** Light orange
* **For:** Salmonella and Shigella spp.
* **Inhibitors:** Brilliant Green, Bile Salts, Citrate
* **Indicators: pH indicator:** Neutral red
* **H2S Indicator:** Ferric ammonium citrate
* **Sulfur Source:** Sodium thiosulfate
115
# **Hektoen Enteric (HEA) Agar**
* **Orginal color:**
* **Selective**
***For:***
***Inhibitors:***
* **Differential:**
***Indicators: pH indicators:***
***H2S indicator:***
***Sulfur Source:***
## Footnote
(DG)
(GNEB)
(BS, C)
(BTB)
(FAC)
(ST)
* **Orginal color:** Dark Green
* **For:** Gram negative Enteric bacilli
* **Inhibitors:** Bile Salts, Citrate
* **Indicators: pH indicator:** Bromthymol Blue (BTB)
* **H2S Indicator:** Ferric ammonium citrate
* **Sulfur Source:** Sodium thiosulfate
116
# **Bismuth Sulfite (BSA) Agar**
* **Selective**
***For:***
***Inhibitors:***
***CHO Incorporated:***
## Footnote
(S spp)
(BMS)
(GL)
* ***For:*** Salmonella spp. (Salm. Typhi has distinct appearance)
* ***Inhibitors:*** Bismuth sulfite
* ***CHO Incorporated:*** Glucose
## Footnote
Salmonella typhi colonies appear as black colonies with silver metallic sheen
117
# **Brilliant Green (BGA) Agar**
* **Selective**
***For:***
***Inhibitors:***
***CHO Incorporated:***
## Footnote
(S spp/S t)
(BG)
(LC)
* ***For:*** Salmonella spp. except for Salmonella typhi
* ***Inhibitors:*** Brilliant green
* ***CHO Incorporated:*** Lactose
## Footnote
*Salmonella spp.*, colonies appear as white colonies resembling a snowflake surrounded by brilliant red medium
118
# **Thiosulfate Citrate Bile Salts Sucrose (TCBS) Agar**
* **Orginal color:**
* **Selective**
***For:***
***Inhibitors:***
* **Differential:**
***Indicators:***
## Footnote
(LG/OG)
(VB spp.)
(TS, C, BS)
(BTB)
* **Original Color:** Light Green/ Olive Green
* **For:** *Vibrio spp.*
* **Inhibitors:** Thiosulfate, Citrate, Bile Salts
* **Indicators:** Bromthymol Blue (BTB)
119
# **Mannitol Salt (MSA) Agar**
* **Orginal color:**
* **Selective**
***For:***
***Inhibitors:***
* **Differential:**
***Indicators:***
## Footnote
(L/SP)
(SP spp.)
(^ s 7.5%)
(PR)
* **Original Color:** Light/ Salmon Pink
* **For:** *Staphylococcus spp.*
* **Inhibitors:** High concentration of salts (7.5%)
* **Indicators:** Phenol Red
120
# **Lowenstein Jensen (LJ) Medium**
* **Orginal color:**
* **Selective**
***For:***
***Inhibitors:***
## Footnote
(LG)
(MB spp.)
(MG)
* **Original Color:** Light Green
* **For:** Mycobacterium spp.
* **Inhibitors:** Malachite Green
121
# **Lowenstein Jensen (LJ) Medium**
**Sputum Sample** needs to be:
## Footnote
(DC/DG)
(DCT)
1. Decongested/ Digested
2. Decontaminated
122
# **Lowenstein Jensen (LJ) Medium**
To dissolve the thick mucus/mucin that might be trapping the bacteria in the sample.
| N-acetyl-L-cysteine (NALC) is usually used
## Footnote
(DC/DG)
Decongested/Digested
123
# **Lowenstein Jensen (LJ) Medium**
To eliminate normal flora that contaminates the sample.
| NaOH is usually used
Decontaminated
124
**Selective Medium for Neisseria spp.**
## Footnote
(N. spp - GNC)
(CHA-AB)
* Neisseria spp., Gram negative cocci
* Usually composed of chocolate agar base with antibiotics
125
# **Selective Medium for Neisseria spp.**
* **Gram + Inhibitor:**
* **Gram - Inhibitor:**
* **Fungal Inhibitor:**
* ***Proteus spp.* Inhibitor**
| **Culture media:** *Thayer Martin Agar (TMA)*
## Footnote
(V,C,N, -)
* Vancomycin
* Colistin
* Nystatin
* no proteus spp. inhibitor
126
# **Selective Medium for Neisseria spp.**
* **Gram + Inhibitor:**
* **Gram - Inhibitor:**
* **Fungal Inhibitor:**
* ***Proteus spp.* Inhibitor**
| **Culture media:** *Modified Thayer Martin Agar (MTMA)*
* Vancomycin
* Colistin
* Nystatin
* Trimethoprim
127
# **Selective Medium for Neisseria spp.**
* **Gram + Inhibitor:**
* **Gram - Inhibitor:**
* **Fungal Inhibitor:**
* ***Proteus spp.* Inhibitor**
| **Culture media:** *Martin-Lewis Agar (MLA)*
* Vancomycin
* Colistin
* Anisomycin
* Trimethoprim
128
# **Selective Medium for Neisseria spp.**
* **Gram + Inhibitor:**
* **Gram - Inhibitor:**
* **Fungal Inhibitor:**
* ***Proteus spp.* Inhibitor**
| **Culture media:** *New York City Agar (NYCA)*
* Vancomycin
* Colistin
* Amphotericin B
* Trimethoprim
129
# **Selective Medium for Neisseria spp.**
* **Gram + Inhibitor:**
* **Gram - Inhibitor:**
* **Fungal Inhibitor:**
* ***Proteus spp.* Inhibitor**
| **Culture media:** *GC-LECT*
* Decreased concentration of Vancomycin; added w/ Lincomycin
* Colistin
* Amphotericin B
* Trimethoprim
130
**CULTURE MEDIA FOR ANTIBIOTIC SUSCEPTIBILITY/SENSITIVITY TESTING (AST)**
## Footnote
(MB, HP spp., MB spp.)
* Most bacteria
* *Haemophilus spp.*
* *Mycobacterium spp.*
131
# CULTURE MEDIA FOR ANTIBIOTIC SUSCEPTIBILITY/SENSITIVITY TESTING (AST)
Mueller Hinton Agar (MHA) and Mueller Hinton Broth (MHB)
Most Bacteria
132
# CULTURE MEDIA FOR ANTIBIOTIC SUSCEPTIBILITY/SENSITIVITY TESTING (AST)
Mueller Hinton with Chocolate Agar Base or Haemophilus Test Medium (HTM) Agar
## Footnote
(fastidious organism – difficult to grow)
*Haemophilus spp.*
133
# CULTURE MEDIA FOR ANTIBIOTIC SUSCEPTIBILITY/SENSITIVITY TESTING (AST)
Middlebrook 7H10 or 7H11 Medium
*Mycobacterium spp.*
134
**Characteristic/Biochemical Culture Media**
## Footnote
(SIMM,MR,VP,SCA,TSIA,LIA,MB,SUB/CUA)
* Sulfide Indole Motility Medium (SIM)
* Methyl Red (MR)
* Voges-Proskauer (VP)
* Simmon Citrate Agar (SCA)
* Triple Sugar Ion Agar (TSI)
* Lysine Iron Agar (LIA)
* Moeller’s Broth
* Stuart’s Urea Broth or Christensen Urea Agar
135
# **Characteristic/ Biochemical Culture Media**
Used for observation of hydrogen sulfide gas production
Sulfide Indole Motility Medium (SIM)
136
# **Characteristic/ Biochemical Culture Media**
there will be presence of black precipitate
| *Sulfide Indole Motility Medium (SIM)*
## Footnote
(HSGP)
hydrogen sulfide gas production
137
# **Characteristic/ Biochemical Culture Media**
can be observed by the appearance of red or pink ring formation
| *Sulfide Indole Motility Medium (SIM)*
## Footnote
(IP)
indole production
138
# **Characteristic/ Biochemical Culture Media**
presence of turbidity or spread of colonies along the side of inoculation
| *Sulfide Indole Motility Medium (SIM)*
## Footnote
(M)
motility
139
# **Characteristic/ Biochemical Culture Media**
Used for the detection of bacterial pathogen that metabolize glucose using the **Mixed Acid pathway**
## Footnote
* Add (__) as reagent
Methyl Red (MR)
140
# **Characteristic/ Biochemical Culture Media**
Used for the detection of bacterial pathogen that metabolizes glucose using the **Butylene Glycol Pathway**
## Footnote
* Add alpha-naptol as reagent
Voges-Proskauer (VP)
## Footnote
**Note:**
* Positive MR = Negative VP
* Negative MR = Positive VP
(to determine what pathway of glucose metabolism)
141
# **Characteristic/ Biochemical Culture Media**
Used for the detection of bacterial pathogen that can utilize citrate as a sole source of carbon
## Footnote
* Green slanted media (original color)
* Positive result (blue)
Simmon Citrate Agar (SCA)
142
# **Characteristic/ Biochemical Culture Media**
Used for the determination of bacterial pathogen’s ability to ferment glucose, sucrose or lactose.
## Footnote
* It can also detect **sulfide
production** (blackening of the agar)
* **gas production** (+ result = bubbles, cracks or spaces on the culture media)
Triple Sugar Ion Agar (TSIA)
143
# **Characteristic/ Biochemical Culture Media**
* **Red/red:**
* **Control**
* **Red/yellow:**
* **Yellow/yellow**:
* **Red/yellow with H2S:**
| ***Triple Sugar Ion Agar (TSI)***
* no sugar fermentation
* control
* Glucose fermented but lactose and sucrose not fermented
* Glucose fermented. Lactose and/or sucrose fermented
* Red/yellow with H2S
144
# ***Triple Sugar Ion Agar (TSI):*** percentage/parts
* **dextrose:**
* **sucrose:**
* **lactose:**
| the "parts" are ***SIMPLIFIED VERSION***
* 0.1% (1 part)
* 1.0% (10 parts)
* 1.0% (10 parts)
145
# **Characteristic/ Biochemical Culture Media**
Used for the determination of bacterial pathogen’s ability to decarboxylate or deaminate lysine.
## Footnote
* It can also detect glucose fermentation, sulfide production and gas production
Lysine Iron Agar (LIA)
146
# **Characteristic/ Biochemical Culture Media**
Used to detect lysine decarboxylation, ornithine decarboxylation, and arginine dihydrolysis
Moeller’s Broth
147
# **Characteristic/ Biochemical Culture Media**
Used to detect bacterial pathogen that hydrolyze urea substrate
Stuart’s Urea Broth or Christensen Urea Agar
