Microbiologic sampling conduction

There are no convenient or reliable functional tests of pantothenic acid status, thus assessment is made by direct measurement of whole blood or urine pantothenic acid concentrations. Urine measurements are perhaps the easiest to conduct and interpret, and concentrations are closely related to dietary intake, Whole blood measurements are preferred to plasma, which contains only free pantothenic acid and is insensitive to changes in pantothenic acid intake. Concentrations of pantothenic acid in aU of the above fluids can be measured by microbiological assay, most commonly using Lactobacillus plantarum. Whole blood must first be treated with an enzyme preparation to release pantothenic acid fi om CoA. Other techniques that have been used to measure pantothenic acid in human samples include radioimmunoassay and gas chromatography, Other techniques that have been developed include gas chromatography-mass spectrometry and a stable isotope dilution assay. CoA and AGP can be measured by enzymatic methods. ... 

The TAC can be conducted using a number of microbiological methods. These are the pour-plate, membrane filtration, and multiple tube methods. The TYMC is conducted by using either the pour-plate or membrane filtration method. The TAC for bioburden is performed by adding 10 g, 10 mL, or 10 units in SCD broth or lactose broth to make 100 mL. Aliquots of this sample preparation are transferred into four standard size (15 x 100 mm) Petri dishes. Into two of the plates 15-20 mL of molten SCD agar is poured, and into the other two the same volume of Sabouraud dextrose agar (SAB) agar is poured. 

Metabolite/residue analysis. Milk, urine and plasma samples were first analyzed by a microbiological cylinder/plate procedure against M.luteus which has a limit of detection of 0.02 ppm. A sub-sample of the milk was prepared for this assay by a centrifugation step followed by a pH adjustment to 8.5. In addition, an HPLC/RAM analysis was conducted after treating another sub-sample with FTSH (10% formic acid, 30% trifluoroacetic acid, 2% sodium chloride, 2N hydrochloric acid) followed by centrifugation to precipitate the proteins. The supernatant was basified and concentrated by C-18 solid phase extraction (SPE) techniques. The HPLC conditions were Column - 20 cm x 4.8 mm C-8 Mobile-phase -linear gradient at 5%/minute from 90 10 O.IM pH 7 phosphate buffer methanol to 20 80 Detectors - UV operated at 214 nm and a radioactivity flow detector operated in the DPM mode. 

Processing should be conducted in a deanroom suite, constructed and operated in accordance with the air cleanliness standards. In order to control the microbiological and particulate cleanliness of the various grades/dasses of operation, the areas should be monitored using various methods, e.g., volumetric air sampling, settle plates, surface sampling (swabs, contact plates). 

At present, four companies provide specialized impedance measurement instrumentation for chnical microbiology or food hygiene applications. BioMirieux manufactures the Bactometer, which can monitor conductance, capacitance, or impedance in up to two temperature-controlled incubators, each containing 256 samples. Malthus Instruments produces... 

This documentation allows the winery to accurately describe what procedures, frequency, and chemicals were used to ensure that operations were conducted in a consistent manner. At the conclusion, representative surfaces should be sampled for microbiological populations and/or excess sanitizer. All data should be collected for future examination and potential changes. These records not only become part of the overall quality points program (Chapter 10) but also will serve as documentation if a legal issue arises. 

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