BACTE-BASICS

1. OXIDASE REAGENT:
- 1% tetramethyparaphenylenediamine dihydrochloride
- Indicator: Bromthymol blue
- Positive: purple
- Use Platinum wire or wooden sticks to transfer colony to avoid false positive
- Initially differentiates groups of gram-neg
- Oxidase neg: Enterobacteriaceae, Stenotrophomonas maltophilia, and Acinetobacter
spp
- Oxidase pos: Pseudomonas spp. and Aeromonas spp
2. Novobiocin: 5ug; S: >16mm
3. Optochin:5ug;
SUSCEPTIBLE if:
- >14 mm ZOI -6mm disk
- >16mm ZOI-10mm disk
4. PYRase:
- Detects the enzyme pyrrolidonyl peptidase/arylamidase or
L-pyroglutamyl-aminopeptidase (PYR)
- Substrate used is pyrrolidonylarylamide or L-pyrrolidonyl-b-naphthylamide
- Developing agent is p-dimethylaminocinnamaldehyde (DMACA)
- PYR (substrate) --PYRase→ beta-naphthylamide
- Beta-naphthylamide + DMACA → BRIGHT RED
- Helps identify S. pyogenes & Enterococcus (+)
5. INDOLE TEST:
- Presumptively identifies E. coli
- Substrate: tryptophan
- Enzyme detected: tryptophanase
- Product detected: indole
- Medium: SIM or peptone water
- Reagents:
- KOVAC’S: (+) PINK TO RED
1. PDAB
2. Isoamyl alcohol
 3. Conc. HCl
- EHRLICH: for anaerobes and nonfermenters
- SPOT INDOLE TEST: (+) BLUE
- p-dimethylaminocinnamaldehyde (DMACA)
- 37% hydrochloric acid (HCl)
- deionized water
6. CITRATE UTILIZATION:
- Medium: simmons citrate agar (slanted)
- Indicator: bromthymol blue
- Positive: blue (alkaline)
- Negative: green (acidic)
7. VOGES PROSKAUER:
Acetoin (from butylene glycol production) --40%KOH + air→ diacetyl
Diacetyl + guanidine in the peptone + 5% alpha naphthol (color intensifier)→ RED
COLOR
- Negative tubes must be held for 1 hr
8. UREASE
- Detects ammonium carbonate
- Urea --urease→ ammonia, water, CO2
- Media used:
- broth (Stuart’s urea broth)
- agar (Christensen’s urea agar)
- Helps identify Proteus spp., Corynebacterium urealyticum and Helicobacter pylori
- Enterobacter spp., exhibits weak urease positive result
9. MOTILITY
- Uses 1% triphenyltetrazolium
10. ONPG
- Positive color is YELLOW
- Detects galactose o-nitrophenyl
HIPPURATE HYDROLYSIS:
- Enzyme: hippuricase
- Substrate: hippurate
- Product: glycine
- Developing color: ninhydrin
 - (+) DEEP PURPLE COLOR:
- Gardnerella vaginalis
- Streptococcus agalactiae
- Campylobacter jejuni
- Listeria monocytogenes

OTHER TESTS:
● O-F TEST:
- pH indicators used:
1. Bromcresol purple: changes from purple to yellow
2. Andrades acid fuchsin: from pale yellow to pink
3. Phenol red: from red to yellow
4. Bromthymol blue: from green to yellow
● DECARBOXYLATION
- Requires an acid environment for activation
- ANAEROBIC
- Medium: Moeller decarboxylase base
- Products produce increase in pH
- (+) purple; (-) yellow
- Arginine → citrulline → ornithine
● DEAMINATION:
- AEROBIC
- End product: phenylpyruvic acid
- Reagent: 10% ferric chloride (detects phenylypyruvic acid)
- (+) GREEN COLOR
- Medium: phenylalanine deaminase agar (PDA)
- (+) proteus
11. BACITRACIN: 0.04IU
12. Sample from a pregnant women suspected of GBS is swabbed between 35 and 37 weeks of
gestation
13. S. anginosus group has a sweet odor of honeysuckle or butterscotch in pure culture or
high conc.
14. Enterococcus grow in extreme environmental conditions: bile, 6.5% NaCl, 45C, alkaline pH
15. Enterococcus exhibits pseudocatalase rxn; weak
16. Aerococcus and Enterococcus both grow in the presence of 6.5%NaCl and are both PYR
positive
17. All viridans are LAP positive and PYR negative
18. Pediococcus can also grow at 6.5%NaCl
19. Viridans is the most common cause of subacute bacterial endocarditis
20. Enterococcus causes UTI, bacteremia, endocarditis, wound infections
21. PYR is more specific to S. pyogenes than is bacitracin
22. Members of anginosus group is VP positive
23. Pediococcus and Leuconostoc can grow in 6.5% NaCl broth when incubated for 24 hrs
24. Bile solubility of S. pneumoniae is due to the enzyme amidase
25. P. aeruginosa: ventilator-associated pneumonia
26. P. aeruginosa: produces mucoid colonies (in px with CF) due to overproduction of alginate
27. Cetrimide agar: for P. aeruginosa
28. P. fluorescens, P. putida, and P. aeruginosa are pigmented and produce acid from xylose
which separates them from other members of Pseudomonas.
29. Pseudomonas are oxidase positive except P. luteola and P. oryzihabitans
30. P. stutzeri is ADH (-) and starch hydrolysis (+); wrinkled “Stutzeri Starch positive”
31. P. mendocina is ADH (+) and starch hydrolysis (-); nonwrinkled
32. P. pseudoalcaligenes is ADH (+) “Pseudo=Positive”
P. alcaligenes is ADH (-)
33. Acinetobacter: 2nd to P. aeruginosa in frequency in all nonfermenters in the lab
34. A. baumannii: saccharolytic → may resemble a lactose-fermenting organism when grown on
MacConkey
35. A. lwoffi: asaccharolytic
36. B. pseudomallei has a characteristic earthy odor on ashdown medium
37. Oligella urethralis: motile
38. Oligella ureolytica: nonmotile
39. Tween 80 stimulates the growth of Psychrobacter
40. Psychrobacter has a characteristics odor of roses in PEA
41. NFO w/ wrinkled colonies:
- P. stutzeri
- B. pseudomallei
- P. oryzihabitans
42. Brevundimonas vesicularis is esculin hydrolysis (+)
43. Brevundimonas diminuta is esculin hydrolysis (-) “Di s’ya positive”
44. Paracoccus yeei is O-shaped on gram stain
45. Chromobacterium violaceum produces a violet pigment called violacein which is
ethanol-soluble & water-insoluble
46. Delftia acidovorans: causes keratitis in soft contact lenses wearers
47. Elizabethkingia meningoseptica: causes meningitis or septicemia in newborns
48. Vibrio parahaemolyticus and V. alginolyticus have an ability to swarm on solid media
49. Vibriostatic compound O/129 or 2,4-diamino-6,7-diisopropylpteridine (sensitive: vibrios &
Plesiomonas; Resistant: Aeromonas)
50. String test: 0.5% sodium deoxycholate
51. Nonhalophilic: V. cholerae & V. mimicus
52. V. cholerae O1: causative agent of cholera
53. V. parahaemolyticus causes “summer diarrhea” in Japan; associated w/ consumption of
oysters
54. V. parahaemolyticus can be found only in coastal or estuarine areas despite being halophilic
55. V. parahaemolyticus is KANAGAWA TOXIN positive in Wagatsuma agar
56. V. vulnificus: 2nd most serious type of Vibrio-associated infections; lactose fermenter
57. V. alginolyticus: least pathogenic; most frequently isolated; strict halophile (able to tolerate
up to 10% NaCl)
58. Glycerol is toxic to most vibrios
59. Vibrios grow as nonlactose fermenters on MAC except for V. vulnificus (differentiate it from
Enterobacteriaceae using oxidase test)
60. TCBS yellow colonies (+):
- V. cholerae
- V. alginolyticus
61. TCBS green colonies (-):
- V. parahaemolyticus
- V. mimicus
- V. vulnificus
62. All Aeromonads can grow from 10-42C and can grow in almost all enteric media
63. Aeromonas caviae: most common cause of GI infections in neonates and pediatric px;
lactose-fermenter → often overlooked as E. coli
64. Media used for high recovery of Aeromonas is SBA + modified CIN II w/ 4ug cefsulodin
instead of 15ug
65. Aeromonas appears pink-centered in CIN I agar
66. Aeromonas: resistant to string test while Vibrios are sensitive
67. Inositol fermentation confirms Plesiomonas
68. Campylobacter jejuni: most common cause of bacterial gastroenteritis worldwide
69. H. pylori: major cause of type B gastritis
70. Campy blood agar: commonly used medium in isolating C. jejuni
71. Campy CVA: for better suppression of fecal biota
72. Use cary-blair for transport of campylobacter
73. Incubation at 42C: optimal growth
74. To observe the motility of Campylobacter, use Brucella or trypticase soy broth; don’t use
distilled water & saline
75. C. jejuni and other enteric campylobacters: runny looking colonies
76. Neisseria species are aerobic and capnophilic; grows anaerobically if alternative electron
acceptors (nitrites) are available
77. Optimal growth of Neisseria: humid environment
78. LOS endotoxin: major virulence factor of neisseria
79. Porin protein: for nutrients; por B is present in N. gonorrhoeae while por A and B are both
present in N. meningitidis
80. Protein II (opacity): for adherence
81. Protein III (reduction modified protein): blocks IgG
82. Strains of Neisseria in asymptomatic men require AHU (arginine, hypoxanthine, uracil)
83. Calcium alginate and cotton swabs inhibit Neisseria. Use dacron or rayon
84. Other organisms that may grow in selective gonococcal medium:
- Acinetobacter spp.
- Capnocytophaga spp.
- Kingella denitrificans
85. CHO utilization for differentiating Neisseria species is done in cystine trypticase agar (TCA)
86. NAATs: uses 1st morning urine; can detect both Neisseria & Chlamydia trachomatis
87. N. gonorrhoeae is inhibited by SPS; add gelatin for neutralization
88. Gamma-glutamyl aminopeptidase: positive for N. meningitidis (differentiates it from N.
gonorrhoeae)
89. ONPG differentiates N. lactamica (+) from N. meningitidis
90. Moraxella produces wagon wheel appearance on choc agar
91. Moraxella: asaccharolytic, DNAse & butyrate esterase positive
92. Tributyrin: substrate used to detect butyrate esterase activity
93. N. cinerea is asaccharolytic and grows on SBA (N. gonorrhoeae does not grow on SBA)
94. N. sicca: dry “breadcrumb-like” colonies
95. N. weaveri: weakly PAD (+)
96. Determines the validity of DNase media:
- M. nonliquefaciens
- M. canis
97. M. catarrhalis & N. mucosa: nitrate reduction positive
98. HACEK: nonmotile & pleomorphic
99. Produces V factor:
- Neisseria
- S. pneumoniae
- S. aureus
100. Capsule of Haemophilus influenzae serotype B is composed of ribose, ribitol, and
phosphate
101. Nontypeable H. influenzae (NTHi) strains are nonencapsulated and cause localized
infections.
102. Haemophilus aegyptius: Koch-Weeks bacillus; causes pink eye
103. H. influenzae biogroup aegyptius causes brazilian purpuric fever
104. H. aegyptius: enriched CHOC agar w/ 1% Isovotalex or vitox
105. H. ducreyi: enriched CHOC or nairobi biplate
106. H. influenzae: mousy/bleach-like odor
107. H. ducreyi: school of fish/fingerprint/railroad tracks pattern on gram stain
108. Haemophilus spp. do not require heme when grown anaerobically but still need NAD
109. H. aphrophilus & H. ducreyi: does not require factor V
1110. H. influenzae and H. haemolyticus both require factor X and V. Differentiate them through
hemolysis on rabbit blood agar. H. haemolyticus produces a B-hemolytic pattern.
111. H. parainfluenzae and H. paraphrophilus both require factor V only. Differentiate them by
using ornithine. H. parainfluenzae is ornithine (+) while H. paraphrophilus is ornithine (-).
112. Porphyrin test: differentiates heme-producing species
113. All haemophilus spp. with “para” are positive for sucrose & porphyrin test
114. All haemophilus spp. are positive for glucose except H. ducreyi.
115. Aggregatibacter aphrophilus: one of the most prevalent species in the HACEK group
involved in endocarditis; causes bone and joint infections
116. Aggregatibacter actinomycetemcomitans is star-shaped and is a major contributor to
periodontitis which can lead to the destruction of the alveolar bone
117. Cardiobacterium hominis: affects aortic valve; forms rosette in gram stain
118. Eikenella corrodens: chlorine bleach-like odor
119. Kingella : most common cause of osteoarthritis infection in children <4 yrs old
- Can grow on MTM
- Resists decolorization
120. Capnocytophaga: gliding motility; thin & fusiform
121. Pasteurella: feline bites; bipolar staining
- P. multocida: most common isolate
122. Brucella: category B select biological agents; biosafety level 3
- Cause undulant fever
- Can reside within mononuclear phagocytic cells
- Specimen of choice: blood or bone marrow
- Can grow on SBA, CHOC and MTM
123. Francisella: category A select biological agent
- Fastidious: needs cystine, cysteine, & thiosulfate
- Strictly aerobic
- Reportable disease
- Other names:
- Rabbit fever
- Deerfly fever
- Lemming fever
- Water rat trappers disease
- Market men;s disease
124. Legionella: survives inside the phagosome
- Resists chlorine → can enter the water supply
- Requires L-cysteine; no L-cysteine → no growth
- Legionnaires disease: urine antigen detection
- Pontiac fever: serology
- Organisms can be found outside of and within macrophages, and segmented
neutrophils
- Isolate using BCYE w/ L-cysteine (ground-glass appearance of the colonies)
- Most are biochemically inert
125. L. micdadei: weakly acid fast in TISSUES
126. Bordetella pertussis is inhibited by fatty acids, metal ions, sulfides, and peroxides which
are all found in media.
- Needs charcoal, blood, or starch which bind & neutralize inhibitory substances
127. Bordetella bronchiseptica causes Kennel cough
128. Specimens for Bordetella are nasopharyngeal aspirates or swabs. Swabs are inserted
through the nares
129. Transport media for Bordetella: “CAR”
- Casamino acid (1% casein hydrolysate)
- Amies with charcoal
- Regan Lowe transport medium
130. Bordetella demonstrates mercury droplets appearance on Regan Lowe medium
131. Nitrate reduction
- Sodium nitrate, HCl, sulfanilamide, N-naphthylenediamine dihydrochloride → RED
COLOR
- Rapid growers can be tested in 2 weeks; slow-growers are tested in 3-4weeks
- Positive:
- M fortuitum
- M. tuberculosis (+) control
- M. kansasii
- M. szulgai
132. TWEEN 80 HYDROLYSIS:
- Lipase splits tween 80 (polyoxyethylene sorbitan monooleate) → oleic acid &
polyoxyethylated sorbitol
- pH indicator used is neutral red
- (+) pink
133. M. chelonae is iron uptake (-); other rapid growers are (+)
134. Positive arylsufatase in 3 days:
- M. fortuitum
- M. chelonae
- M. xenopi
- M. triviale
135. Positive arylsulfatase in 14 days: “SM”
- M. marinum
 - M. szulgai
**enzyme breaks down phenolphthalein disulfate into phenolphthalein (which forms a red
color in the presence of sodium bicarbonate) and other salts
136. PYRAZINAMIDASE
- PZA → pyrazinoic acid & ammonia
137. MAC is tellurite reduction positive (3-4 days)
- Distinguishes it from other nonchromogenic
138. Heat stable catalase (+) ; heated to 68°C for 20 minutes
- MAC
- M. ulcerans
- M. genavense
- M. kansasii
- M. xenopi
- M. simiae
**SEMI-QUANTITATIVE: measures the height of the bubbles (</> 45mm)
139. Cervical lymphadenitis in children: MAC (Mahon)
- 2nd to MAC is M. chelonae
- M. scrofulaceum is the number 1 cause of cervical lymphadenitis in children (Ciulla)
140. Rapid growing localized cutaneous infection: M. fortuitum
141. Growth at 30-32C (poor growth at 37C)
- M. haemophilum
- M. ulcerans
- M. marinum
142. M. xenopi: grows at 42C
143. Serum albumin agar media: 7H10 & 7H11
144. Use 5% oxalic acid to decontaminate specimens contaminated w/ P. aeruginosa
145. Most commonly used is NALC or DTT + NaOH
146. Borrelia burdorgferi sensu lato complex: causes Lyme borreliosis
147. Leptospires are obligate aerobes
148. Media for leptospires:
- EMJH
- Fletcher semisolid medium
- Stuart liquid medium
149. Weil disease: severe systemic disease caused by leptospira interrogans serovar
icterohaemorrhagiae
150. Endemic relapsing fever by Borrelia recurrentis is caused by soft ticks from the genus
ornithodoros
Epidemic relapsing fever: louse
151. Borrelia burdorgferi sensu lato complex: by hard tick (Ixodes)
152.. Treponemes: graceful flexuous movements in liquid
153. Yaws: by Treponema pallidum subsp. pertenue
154. Pinta: by T. carateum
155. Bejel: by Treponema pallidum subsp. endemicum
156. Mycoplasma & Ureaplasma require cholesterol and fatty acids for growth
157. M. hominis: fried-egg appearance; no fried-egg appearance under anaerobic conditions
158. M. pneumoniae: causes walking pneumonia
159. Ureaplasma urealyticum: causes nongonococcal urethritis in men
160. Transport medium for Mycoplasma and Ureaplasma:
- SP4 (sucrose phosphate buffer, Mycoplasma base, horse serum, neutral red)
- Shepherd 10B broth or 2SP
- 10B broth: for genital specimens of Ureaplasma ueralyticum
161. M. pneumoniae & M. genitalium requires glucose for growth
162. M. hominis requires arginine
163. Ureaplasma requires urea; dies rapidly when urea is depleted
164. SPS is inhibitory to Mycoplasma; add gelatin for neutralization
165. Mycoplasma stain: Dienes or methylene blue stain
166. Inhibited by SPS:
- G. vaginalis
- Neisseria
- Streptobacillus moniliformis
- Peptostreptococcus anaerobius
- Mycoplasma
167. Vibrio and campylobacter: 42C optimum temp
168. H. pylori: stuart medium for transport
169. Shigella sonnei
- Lactose fermentation positive (late)
- ONPG positive
 - Ornithine positive
170. H2S positive in TSI: SPACEd
171. H2S positive in LIA: SACEd
172. Salmonella is H2S positive except Paratyphi A
173. B. anthracis:
- Medusa head
- Bamboo pole appearance
- Beaten egg white
- NONMOTILE (unique)
- Causes malignant pustule (black eschar)
174. VKC REACTION

Vancomycin Kanamycin Colistin

Prevotella R R V

Porphyromonas S R R

Fusobacterium R S S

C. ureolyticus R S S

Bacteroides R R R

175. Arcanobacterium haemolyticum and C. perfringens:

- Reverse CAMP rxn (+)
176. Erysipelothrix: gram positive and H2S (+)
- Causes erysipeloid
177. Pyrazinamidase:
- M. marinum (+); M. kansasii (-)
- M. tuberculosis (+); M. bovis (-)
178.

Hydrolysis of tween 80 Urease

M. gordonae + -

M. scrofulaceum - +

179. M. kansasii: hydrolysis of tween 80 is positive in 6 h rs

180. Prevotella grows on KVLB but not on BBE
181. C. ureolyticus does not grow on both KVLB and BBE
182. C. sordellii: the only urease positive clostridium
183. C. difficile has glutamate dehydrogenase
184.
MOTILE:
- C. botulinum
- C. tetani
- C. difficile
NONMOTILE:
- C. perfringens
185. Positive porphyrin test:
- H. parainfluenzae
- H. aphrophilus
- H. paraphrophilus
186. Transport for chlamydia:
- Sucrose phosphate glutamate buffer
187. Klebsiella granulomatis: causes granuloma inguinale
188. Lecithinase test/nagler’s test
189. Coxiella burnetii: no rashes
- Not arthropod-borne
190. Chlamydophila pneumoniae: associated w/ coronary heart disease
191. SPECIMEN FOR LEPTOSPIROSIS
First week of leptospirosis: blood
After the1st week: urine
192. PAD (+) is green
- 10% ferric chloride
193. Moeller’s medium: for decarboxylation
194. Paper strip test: 5% lead acetate
- For hydrogen sulfide production
195. The resolving power of most light microscopes allows bacterial cells to be distinguished
from one another but usually does not allow bacterial structures, internal or external, to be
detected
196. Phase-contrast microscopy does not use a fixed smear preparation, but instead is used to
view organisms and other cells in a wet preparation or wet mount
197. Photobleaching or fading is the permanent loss of fluorescence as a result of chemical
damage to the fluorochrome. Quenching is a result of the transfer of the light energy to nearby
molecules in the sample such as free radicals, salts of heavy metals, or halogens

198. Acridine orange binds to nucleic acids

199. Immunofluroescence: used to directly examine patient specimens for bacteria that are
difficult or slow to grow (e.g., Legionella spp., Bordetella pertussis, and Chlamydia trachomatis)
or to identify organisms already grown in culture
200. TEM passes the electron beam through objects and allows visualization of internal
structures. SEM uses electron beams to scan the surface of objects and provides
three-dimensional views of surface structures
201. Schaedler agar: Nonselective medium for the recovery of anaerobes and aerobes
Selective for Campylobacter spp. and Helicobacter spp.
202. Listeria monocytogenes and Yersinia enterocolitica, can grow at 4°C to 43°C but grow best
at temperatures between 20° and 40°C. Cold enrichment has been used to enhance the
recovery of these organisms in the laboratory
203. Most clinically relevant bacteria prefer a near-neutral pH range, from 6.5 to 7.5.
204. catalase reaction differentiates Listeria monocytogenes and corynebacteria
(catalasepositive) from other gram-positive, non–spore-forming bacilli

PS: sabihin nyo na lang sakin if may mali hehe thank you!