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Media Background Incubation Time and Maximum stacking height? Analysis SOP Tar...

Media Background Incubation Time and Maximum stacking height? Analysis SOP Target Organism Organism Background Media Miscellaneous (Pour/Spread, Selectivity, etc.) Temperature Plates inverted? Plates bagged? VIS-NC-002 12 plates max What is a standard plate count? Pour plate SPC 30 72 ± 3 hours It is an estimate of microorganisms in a Milk Plate Count Agar Standard Plate Count Plates inverted sample (MPCA) Non-selective 30 ± 1 C Not bagged What is the difference between selective QA-0015-0201 APC-PSYCHR Pour plate Standard Methods Agar 10 days and non-selective? 12 plates max (SMA) Non-selective does not have a target What temperature should the media be Bacteria that can grow at temperatures 7C - the lower incubation organism and allows a variety of Psychrotrophs when poured? Plates inverted less than or equal to 7 C Milk Plate Count Agar temperature of this test organisms to grow, selective has properties 40-45 C, if the media temperature is (MPCA) makes it selective for (media ingredients, temperature, higher than this range there is a reduced Bagged *for client butter samples* psychrotrophic organisms aerobic/anaerobic environment) that inhibit recovery of psychrotrophic organisms non-target organisms and "selects" for the target organism How should each petri dish be labeled? Sample label with complete Sample ID and dilution QA-0015-0240 12 plates max What is the purpose of placing petri What is a standard plate count? SPC 32 Pour plate 48 ± 3 hours dishes in bags during incubation? It is an estimate of microorganisms in a Standard Methods Agar Standard Plate Count Plates inverted To prevent the media from drying out under sample (SMA) Non-selective 32 ± 1 C high temperatures or long incubation time, Not bagged or prevent "spreader" colonies Point #1 How long after pipetting a sample should agar be poured? 20 minutes Potato Dextrose Agar What needs to be checked before QA-0035-0501 Yeast: single-celled eukaryotes (PDA) with tartaric acid 6 plates max pouring media? Pour Plate Pour plate ISO 6611 Media expiration, temperature (45-48 C) for (TTA) 120 ± 6 hours YM - Mold: filamentous multicellular eukaryotes YEA inverted most methods Yeasts and Mold Selective PDA not inverted Yeast Extract Glucose TTA lowers pH (pH range 3.4-3.6) to 22-25 C How long is media usable after being Both can be found in soil, air, in food, on Chloramphenicol Agar select for yeasts and molds melted? food, and processing environments Bagged (YEA) 4 hours SMEDP 8.112/8.113 Yeast Extract Glucose QA-0035-0540 Spread Plate Chloramphenicol Agar Spread plate 6 plates max Yeast: single-celled organisms (YEA) 5 ± 0.5 days YM - Yeasts and Mold Selective Not inverted Mold: filamentous multicellular organisms Potato Dextrose Agar chloramphenicol eliminates bacteria and 25 ± 1 C (PDA) with tartaric acid not yeasts and molds Bagged (TTA)  Media Information Incubation Time and Maximum Stacking Height? Analysis SOP Target Organism Organism Background Media Miscellaneous (Pour/Spread, Selectivity, etc.) Temperature Plates inverted? Plates bagged? QA-0015-0205 APC-THERMP 48 ± 2 hours Pour plate 12 plates max Organisms that can survive high Standard Methods Agar 55 ± 1 C temperatures; they are naturally occuring Thermophiles (SMA) Capping is optional, and is used to Higher incubation Inverted in soils and can still be present after prevent spreading temperature makes this test thermal processing *We do not cap this test at our location* selective for thermophilic Bagged organisms How do you properly homogenize the sample with the agar once it has been added? In a figure 8 motion, distribute the sample Pour plate within the agar until it is completely QA-0017-0240 Gram-negative bacilli that ferment incorporated and there are no visible glucose to acid and are oxidase negative 12 plates max Capped with 5-10 mLs to prevent streaks in the agar EB-QN 24 ± 2 hours Violet Red Biole Glucose spreading Enterobacteriaceae (EB) Indicator orgranism - a microorganism Inverted Agar (VRBG) What is the maximum volume we can that indicates the presence of pathogens, 37 ± 1 C Selective pipette onto a spread plate? or disease-causing organisms, in water, Not bagged Bile salts and crystal violet inhibit bile- 0.4 mLs food, or the environment sensitive and gram-positive bacteria How do you pipette 1 mL over 3 spread plates? 0.3 mLs/0.3 mLs/0.4 mLs Pour plate How do you properly spread sample on QA-0020-0303 Gram-negative rod-shaped bacteria that TSA is non-selective and allows a wide pre-poured agar plates? ferment lactose variety of organims to grow 12 plates max TSA-VRB Point #2 Trypticase Soy Agar (TSA) Using a L-shaped spreader, spread the 24 ± 2 hours sample towards the edge of the plate Coliforms Indicator orgranism - a microorganism Wait time for capping: 2 hrs ± 30 minutes Inverted without touching the petri dish that indicates the presence of pathogens, Violet Red Bile Agar (VRB) Capped with 8-10 mLs of VRB 32 ± 1 C or disease-causing organisms, in water, Not bagged How are Enterobacteriaceae and food, or the environment VRB is selective - coliforms related? Bile salts and crystal violet inhibit bile- Coliforms are a subgroup of the sensitive and gram-positive bacteria Enterobacteriaceae family; all Coliforms are EBs, but not all EBs are coliforms Pour plate How do we distinguish VRB and VRBG QA-0045-0601 *KF media solidifies quickly. If you do not jars in the lab? have a swirler, you need to swirl every Indicator orgranism - a microorganism 12 plates max Reagent label has media name, and VRBG couple of plates* 48 ± 2 hours that indicates the presence of pathogens, is highlighted or marked with a line to KF Enterococci Kenner Fecal Agar (KF) differentiate between the two or disease-causing organisms, in water, Inverted Selective 35 ± 1 C food, or the environment Sodium azide selects for enterococci, Not bagged How do we distinguish PDA plates from triphenyltetrazolium chloride turns colony other media in the lab? pink Reagent label has media name, and since the agar color is similar to other pre-poured plates media draws a line along each stack of PDA plates Spread plate QA-0030-0401 9 plates max CP Staph Selective 48 ± 2 hours Not inverted Coagulase positive Gram-positive, catalase positive Lithium chloride and potassium tellurite to Baird-Parker Agar (BP) *plate readers invert the plates after 24 Staphylococci microorganism inhibit non staph, Tellurite and egg yolk to 35 ± 1 C hour incubation* provide differentiation, tellurite turns the colony black and egg yolk is hydrolyzed Not bagged to create the clear zone (halo)