The document outlines methodologies for detecting Salmonella in various food samples, primarily focusing on incubation times, culture media, and biochemical confirmation tests. It includes specific protocols for raw flesh foods, highly contaminated foods, and animal feeds, as well as details on using chromogenic media to distinguish microorganisms based on enzymatic reactions. Additionally, it presents a range of biochemical tests and the Himedia product line for microbiological media tailored to different organisms.

13. IISSOO SSttaannddaarrdd 66557799
DDeetteeccttiioonn ooff SSaallmmoonneellllaa
Day
1
BPLS Agar
modified
24-48 h /
35-37°C
Any other
Salmonella
Agar
Day
2
Day
3
Day
4
Day
5
Day
6
For confirmation take from each plate 5 suspected colonies
and streak on Nutrient agar 18 - 24 h / 35 - 37 °C
Biochemical / Serological Confirmation Tests
TSI / Urea / Lysin / b-Gal / VP / Indol
10 ml BPW to
100 ml SC Broth
24 h at 35 - 37 °C 48 h
0.1 ml BPW to
10 ml RVS Broth
24 h / 42 °C
BPLS Agar
modified
24-48 h /
35-37°C
Any other
Salmonella
Agar
BPLS Agar
modified
24-48 h /
35-37°C
Any other
Salmonella
Agar
25 g/ml test sample
in 225 ml BPW
16 - 20 h / 35 - 37 °C

14. FDA-Bacteriological Analytical MMaannuuaall 88tthh EEdd.. 11999955
CChhaapptteerr 55.. SSaallmmoonneellllaa mmeetthhoodd ffoorr rraaww fflleesshh
ffooooddss ,, hhiigghhllyy ccoonnttaammiinnaatteedd ffooooddss aanndd aanniimmaall ffeeeeddss
Day
1
Day
2
Day
3
Day
4
Day
5
Day
6
25 g or 25 ml sample
in 225 ml Lactose Broth
1 ml Lactose Broth to
10 ml TT Broth
24 ± 2 h at 43 ± 0.2 °C
Subculture from RV and TT Broth to a set of 3 plates each
Bismuth Sulfite Agar
if negative after 24 h
Take from each plate suspected colony and streak on Triple
Sugar Iron Agar ( TSI ) and Lysin Iron Agar ( LIA ) 24± 2 h at 35
°C
Further Biochemical / Serological Confirmation Tests
24± 2 h
at 35 °C
Bismuth Sulfite
Agar # 5418
24± 2 h
at 35 °C
0.1 ml Lactose Broth to
10 ml RV Broth
24 ± 2 h at 42 ± 0.2 °C
22 - 26 h / 35 °C
XLD Agar
24 ± 2 h
at 35 °C
Hektoen Agar
24 ± 2 h
at 35 °C

15. Ba FDA-Bacctteerriioollooggiiccaall AAnnaallyyttiiccaall MMaannuuaall 88tthh EEdd.. 11999955
CChhaapptteerr 55.. SSaallmmoonneellllaa mmeetthhoodd ffoorr ootthheerr ffooooddss tthhaann
rraaww fflleesshh ffooooddss ,, hhiigghhllyy ccoonnttaammiinnaatteedd ffooooddss aanndd
aanniimmaall ffeeeeddss
1 ml Lactose Broth to
10 ml TT Broth
24 ± 2 h at 35 °C
1 ml Lactose Broth to
10 ml SC Broth
24 ± 2 h at 35 °C
Day
1
25 g or 25 ml sample
in 225 ml Lactose Broth
22 - 26 h / 35 °C
Day
2
Day
3
Day
4
Day
5
Subculture from SC and TT Broth to a set of 3 plates each
if negative after 24 h
Take from each plate suspected colony and streak on Triple Sugar Iron
Agar ( TSI ) and Lysin Iron Agar ( LIA ) 24± 2 h at 35 °C
Day
6
Further Biochemical / Serological Confirmation Tests
Bismuth Sulfite Agar
24± 2 h
at 35 °C
Bismuth Sulfite Agar
# 5418
24± 2 h
at 35 °C
XLD Agar
24 ± 2 h
at 35 °C
Hektoen Agar
24 ± 2 h
at 35 °C

63. Stomacher

71. Kultur media
Di dalam kapsul

72. Cara Pembuatan
Suspending
Heating
Digesting
Homogenating
for autoclave
Homogenating
after autoclave

73. Cara Pembuatan
Suspending
Heating
Digesting
Homogenating
for autoclave
Homogenating
after autoclave

74. Cara Pembuatan
Suspending
Heating
Digesting
Homogenating
for autoclave
Homogenating
after autoclave

75. Cara Pembuatan
Suspending
Heating
Digesting
Homogenating
for autoclave

76. Cara Pembuatan
Suspending
Heating
Digesting
Homogenating
for autoclave
Homogenating
after autoclave

77. Cara Pembuatan
Suspending
Heating
Digesting
Homogenating
for autoclave
Homogenating
after autoclave

83. Kata Chromogenic berasal dari
kata Chronos yang berarti warna
dan genesis yang berarti
pembuatan.
Jika disatukan mungkin kata
chromogenic berasal dari kata
proses pembuatan warna atau
proses pembuatan pigment.

84. Media mikrobiologi Chromogenik
adalah media mikrobiologi yang
digunakan untuk membedakan
satu mikroorganisme dengan
yang mikroorganisme lain
dengan adanya perubahan warna
yang jelas dengan bantuan
substate chromogenic.

85. Penggunaan Chromogenic
substrate seperti ONPG, X-Gal,
X-Glu bersama dengan
selektifitas dan sensitifitas
media merupakan dasar dari
prinsip sederhana dari media
mikrobiologi chromogenic.

86. Enzimatic Reaction
ß-D-Galactosidase
– 95 % of Coliforms
ß-D-Glucuronidase
–94-97 % of E.coli
Tryptophanase
– 99 % of E.coli are Indole positive

87. Table : Summary of possible enzyme activities, chromogenic substrates, and
selectivity system for microorganisms
Species Enzyme Substrate Selective Agents
Bacillus cereus β-glucosidase, specific Phospholipase C indoxyl-β-glucopyranoside, indoxyl-myo-
inositol-1-phosphate
polymyxin B
Campylobacter n/a n/a deoxycholate, cefoperazone,
amphotericin B
Candida Βacetylgalactosaminidase, alkaline phosphatase indoxyl-N-acetyl-β-D-glucosaminide,
indoxyl-phosphate
Chloramphenicol, Gentamicin
Clostridium perfingens β-glucosidase (plus sucrose fermentation) indoxyl-β-D-glucoside D-cycloserine, polymixin B
Coliforms/E. coli β-glucuronidase,
β-galactosidase
indoxyl-β-glucuronide, Indoxyl-β-
galactoside
bile salts, tergitol 7R, SDS,
novobiocin, cefsulodin
Cronobacter (E.sakazakii) α-glucosidase indoxyl-α-D-glucoside deoxycholate, crystal violet,
sodium thiosulfate
E. coli O157 β-glucosidase, α-galactosidase indoxyl-β-D-glucuronide, indoxyl-α-
galatoside
bile salts, SDS, crystal violet,
potassium tellurite, novobiocin,
cefixime

88. Enterococci β-D-glucosidase indoxyl-β-glucoside sodium azide, polysorbate 80
Extended Spectrum β-Lactamase
Enterobacteria (ESBL)
β-D-glucosidase indoxyl-β-glucoside cefpodoxime, cefotaxime,
ceftazidime
Klebsiella β-D-ribofuranosidase, β-D-glucosidase indoxyl-β-D-ribofuranoside,
indoxyl-β-D-glucoside
bile salts, SDS, carbenicillin
Listeria spp. β-glucosidase indoxyl-β-glucoside lithium chloride, ceftazidime
amphotericin B, nalidixic acid,
polymyxin B
L. monoctogenes Phosphatidylinositol-specific
Phospholipase C, β- glucosidase
indoxyl-β-glucoside, indoxyl-myo-
inositol-1-phosphate
lithium chloride, ceftazidime,
amphotericin B, nalidixic acid,
polymyxin B
Pseudomonas β-Alanyl arylamidase 7-Amido-1-pentyl-phenoxazin-
3-one
cetrimide

89. Salmonella α-galactosidase, lipase indoxyl-α-galactoside, indoxyl-fatty acid ester sodium deoxycholate
MRSA (Methicillin Resistant
Staphylococcus aureus)
α-glucosidase indoxyl-α-D-glucopyranoside methicillin, high concentration of sodium
chloride
Staphylococcus aureus α-glucosidase, phosphatase,
deoxyribonuclease
indoxyl-α,D-glucoside, phenolphthalein
phosphate, indoxyl-phosphate
tellurite, lithium chloride
Streptococci β-glucuronidase indoxyl-β-glucuronide sodium azide
UTI (Urinary Tract Infections) β-glucosidase,
β-galactosidase
indoxyl-β-glucopyranoside,
indoxyl-β-galactoside
Vibrio β-glucosidase, β-galactosidase indoxyl-β-glucopside, indoxyl-β-galactoside,
indoxyl-β-galactoside
high concentration of sodium chloride,
sodium thiosulphate, sodium citrate,
sodium cholate
VRE (Vancomycin Resistant
Enterococci)
α-glucosidase,
β-glucosidase,
β-galactosidase
indoxyl-α-glucopyranoside, indoxyl-β-
glucopyranoside, indoxyl-β-galactoside
vancomycin
Yeasts and Moulds β-N-acetylgalactosaminidase, β-
xylosidase
indoxyl-N-acetyl-β-D-glucosaminide, indoxyl-
β-D-xyloside
oxytetracycline

90. Table: HiMedia product line of chromogenic media according to organisms detected
Organisms Brand Cat. No. Media
Bacillus cereus HiMedia M1651 HiCrome™ Bacillus Agar*
E. coli HiMediaHiMedia M1295
M1488
HiCrome™ E. coli Agar B*
HiCrome™ ECD Agar with MUG*
E. coli & Coliforms HiMedia
HiMedia
HiMedia
HiMedia
M1300
M1293
M1294
M1826
HiCrome™ Coliform Agar*
HiCrome™ ECC Agar*
HiCrome™ ECC Selective Agar*
HiCrome™ Rapid Coliform Broth*
E. coli 0157:H7 HiMedia
HiMedia
HiMedia
HiMedia
M1574
M1575
M1598
M1340
HiCrome™ EC O157 Agar*
HiCrome™ EC O157:H7 Selective Agar, Base*
HiCrome™ Enrichment Broth Base for EC O157:H7*
HiCrome™ Mac Conkey Sorbitol Agar*
Thermotolerant E. coli HiMedia M1571 HiCrome™ m-TEC Agar*
Enterobacter sakazakii (Cronobacter spp.) HiMedia
HiMedia
MV1577
MV1641
HiCrome™Enterobacter Sakazakii HiVeg Agar*
HiCrome™Enterobacter Sakazakii HiVeg Agar, Modified*
Enterococci HiMedia
HiMedia
M1376
M1414
HiCrome™ Enterococci Broth*
HiCrome™ Enterococci Agar*
Enterococcus faecium HiMedia M1580 HiCrome™Enterococcus faecium Agar Base*

91. Table: HiMedia product line of chromogenic media according to organisms detected
Klebsiella HiMedia MV1573 HiCrome™ Klebsiella Selective Agar Base*
Listeria HiMedia M1417 HiCrome™ Listeria Agar Base, modified*
Proteus, enteropathogenic gram-positive
organisms
HiMedia M1418 HiCrome™ UTI Agar, modified*
Salmonella HiMedia
HiMedia
HiMedia
HiMedia
HiMedia
M1393
M1633
M1634
M1296
M1466
HiCrome™ MM Agar*
HiCrome™ RajHans Medium, Modified*
HiCrome™ Salmonella Agar*
HiCrome™ Salmonella Agar, Improved*
Staphylococcus aureus HiMedia M1468 HiCrome™ Aureus Agar Base*
Methicillin-resistant Staph. aureus HiMedia M1674 HiCrome™ MeReSa Agar Base*
Vibrio HiMedia MV1682 HiCrome™ Vibrio Agar*
Yeasts and fungi HiMedia M1467 HiCrome™ OGYE Agar Base*

92. Table : Biochemical Tests on Disks and Strips
Cat. No. Product Description
DD015-1VL Bacitracin Disks Used in the presumptive identification of group A β-hemolytic streptococci.
DD024-1VL Bile Esculin Disks For rapid detection of esculin hydrolysis in the presence of bile for differentiating group D streptococci from
non-group
D streptococci.
DD040-1VL DMACA Indole Disks Used for the Indole test to determine the ability of an organism to split tryptophan into indole and α-
aminopropionic acid.
DD035-1VL Hippurate Disks Recommended in qualitative procedures to detect organisms possessing the enzyme hippurate hydrolase.
DD019-1VL Kovac’s Reagent Strips Used to determine the ability of microorganisms, primarily Enterobacteriaceae, to split indole from the tryptophan
molecule.
DD042-1VL Nitrate Reagent Used to detect an organism’s ability to reduce nitrate.
DD008-1VL ONPG Disks Used to detect the presence of β-galactosidase, an enzyme found in lactose-fermenting organisms.
DD009-1VL Optochin Disks Optochin (ethyl hydrocuprein hydrochloride) is inhibitory for pneumococcal growth whereas other streptococci show
good growth or a very small zone of inhibition.
DD018-1VL Oxidase Disks
DD036-1VL Sterile Disks Used to test a variety of antibiotics, carbohydrates, substrates, and antiseptics on bacteria in petri dishes. Soak a disk in
a
solution or apply some solution on the disks.
DD022-1VL X + V Factor Disks Haemophilus species requiring both X and V factors exhibit growth only in the vicinity of the X +V factor disks.
DD021-1VL V-Factor Disks

97. What Does ATP Detect ?
Total Organic Living Soil :
-Microbial
-Animal
-Plant
-Human
Information :
Total Microbial Contamination
+
Total Food Debris

98. What Does ATP Detect ?

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