rapid microbiological methods
DESCRIPTION
Informasi tentang Rapid Microbiological Methods dan Berbagai jenis yang banyak digunakan di Indonesia.TRANSCRIPT
ISO Standard 6579ISO Standard 6579
Detection of SalmonellaDetection of Salmonella
Day1
BPLS Agarmodified24-48 h /35-37°C
Any otherSalmonella
Agar
Day2
Day3
Day4
Day5
Day6
For confirmation take from each plate 5 suspected colonies and streak on Nutrient agar 18 - 24 h / 35 - 37 °C
Biochemical / Serological Confirmation TestsTSI / Urea / Lysin / -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 Agarmodified24-48 h /35-37°C
Any otherSalmonella
Agar
BPLS Agarmodified24-48 h /35-37°C
Any otherSalmonella
Agar
25 g/ml test samplein 225 ml BPW
16 - 20 h / 35 - 37 °C
FDA-Bacteriological Analytical Manual 8th Ed. 1995FDA-Bacteriological Analytical Manual 8th Ed. 1995Chapter 5. Salmonella method for raw flesh Chapter 5. Salmonella method for raw flesh
foods , highly contaminated foods and animal feedsfoods , highly contaminated foods and animal feeds
Day1
Day2
Day3
Day4
Day
5
Day6
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
Bismuth Sulfite Agar
24± 2 h at 35 °C
Bismuth Sulfite Agar # 5418
24± 2 h at 35 °C
if negative after 24 h
Subculture from RV and TT Broth to a set of 3 plates each
1 ml Lactose Broth to 10 ml TT Broth
24 ± 2 h at 43 ± 0.2 °C
0.1 ml Lactose Broth to 10 ml RV Broth
24 ± 2 h at 42 ± 0.2 °C
25 g or 25 ml samplein 225 ml Lactose Broth
22 - 26 h / 35 °C
XLD Agar
24 ± 2 h at 35 °C
Hektoen Agar
24 ± 2 h at 35 °C
FDA-Bacteriological Analytical Manual 8th Ed. 1995FDA-Bacteriological Analytical Manual 8th Ed. 1995Chapter 5. Salmonella method Chapter 5. Salmonella method for other foods thanfor other foods than
raw flesh foods , highly contaminated foods andraw flesh foods , highly contaminated foods andanimal feedsanimal feeds
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
Day1
25 g or 25 ml samplein 225 ml Lactose Broth
22 - 26 h / 35 °C
Day2
Day3
Day4
Day5
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
Day6
Further Biochemical / Serological Confirmation Tests
Bismuth Sulfite Agar
24± 2 h at 35 °C
Bismuth Sulfite Agar # 5418
24± 2 h at 35 °C
if negative after 24 h
Subculture from SC and TT Broth to a set of 3 plates each
XLD Agar24 ± 2 h at 35 °C
Hektoen Agar
24 ± 2 h at 35 °C
Stomacher
Kultur mediaDi dalam kapsul
Cara Pembuatan
Suspendi
ng HeatingDigesting
Homogenating for autoclave
Homogenating after autoclave
Cara Pembuatan
Suspendi
ng HeatingDigesting
Homogenating for autoclave
Homogenating after autoclave
Cara Pembuatan
Suspendi
ng HeatingDigesting
Homogenating for autoclave
Homogenating after autoclave
Cara Pembuatan
Suspendi
ng HeatingDigesting
Homogenating for autoclave
Cara Pembuatan
Suspendi
ng HeatingDigesting
Homogenating for autoclave
Homogenating after autoclave
Cara Pembuatan
Suspendi
ng HeatingDigesting
Homogenating for autoclave
Homogenating after autoclave
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.
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.
Penggunaan Chromogenic substrate seperti ONPG, X-Gal, X-Glu bersama dengan selektifitas dan sensitifitas media merupakan dasar dari prinsip sederhana dari media mikrobiologi chromogenic.
Enzimatic Reaction
ß-D-Galactosidase– 95 % of Coliforms
ß-D-Glucuronidase–94-97 % of E.coli
Tryptophanase– 99 % of E.coli are Indole positive
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
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
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
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*
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*
Table: HiMedia product line of chromogenic media according to organisms detected
Table : Biochemical Tests on Disks and StripsCat. 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
What Does ATP Detect ?
Total Organic Living Soil :
-Microbial-Animal-Plant-Human
Information :
Total Microbial Contamination +
Total Food Debris
What Does ATP Detect ?
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