Culture-based technique

Overview of Mass Spectrometric Based Techniques Applied in the Cultural Heritage Field... 

A recently published book provides an excellent survey of issues that relate to contamination with endotoxins (present in both viable and nonviable bacteria), their released cell wall constituents, and also viable bacteria in the pharmaceutical industry [1]. It is important to know both the content of the work environment (e.g., indoor air) and the pharmaceutical products themselves. The former provides information on possible sources of microbial contamination and the latter the purity of the final commercial product (or precursors in various stages in its preparation). In some cases it is vital to know the actual bacterial species involved in contamination culture-based methods are standard microbiological techniques which were the focus of Jimenez [1] and thus will not be discussed further. Any contamination (e.g., with endotoxins), regardless of the species of origin, is of utmost of importance (e.g., in determining the safety of a batch of antibiotics to be administered intravenously). This is determined optimally by non-culture-based methods. 

In plant cell cultures, shake flask culture is an indispensable stage of cultivation. Investigations in a shake flask are very essential and critical to bioprocess scale-up and optimization. We have developed a simple and convenient technique based on the principle of the Warburg manometric method to measure 02 uptake rate (OUR) and C02 evolution rate (CER) of suspended cells in a shake flask culture. This technique has been successfully applied to suspension cultures of Panax notoginseng cells, and some important bioprocess parameters, such as OUR, CER, respiratory quotient (RQ), SOUR and specific CER (SCER), were quantitatively obtained [99]. As long as the environment temperature is strictly controlled to within an error of 0.1 °C, the measuring system is accurate and reproducible, is easy to operate, is economical, and is also able to treat many samples simultaneously. 

TIR-based techniques that use fluorescence transduction (TIRF) have been used for the majority of the applications discussed above. These platforms, while not currently the gold standard for measurement of many target analytes, do offer advantages over current technologies, such as cell culture, chromatography, and the enzyme-linked immunosorbant assay (ELISA). Advantages include the ability to perform faster, more sensitive, multiple analyte, and real-time measurements. 

Early studies of marine bacteria via culture-based methods indicated a low abundance in seawater (Sverdrup et ah, 1942 ZoBeU, 1941), and that their assemblages were not diverse (Waksman, 1934). Rehable enumeration techniques developed in the late 1970s indicated that typical bacterial abundances in marine surface waters exceeded colony-forming unit (cfu) estimates by over two orders of magnitude (Hobbieeia/., 1977), with abundances of typically 10 cells in surface waters. 

The use of two-dimensional gel electrophoresis for differential analysis in proteomics was revolutionized by the introduction of 2-D fluorescence difference gel electrophoresis (2-D DIGE). This fluorescence-based technique allows the use of multiplexed samples and an internal standard that virtually eliminates gel-to-gel variability, resulting in increased confidence that differences found between samples are due to real induced changes, rather than inherent biological variation or experimental variability. 2-D DIGE has quickly become the gold standard for gel-based proteomics for studying tissues, as weU as cell culture and bodily fluids such as serum or plasma. 

New studies show thaC culture-based methods (such as viable plate count) do not produce the same results as non-culture-based methods such as epifluorescence microscopy techniques. Therefore, it is always good to use some of these methods together or use better methods such as genetic engineering techniques to distinguish the presence of bacterial populations. ... 

Several recent studies have demonstrated the importance of understanding the general microbial communities of which aquaculture pathogens are members. For these studies, the culture-based approaches have severe limitations since only 0.01-10 % of bacteria in marine systems can be cultured by conventional techniques (Connon and Giovannini, 2002 Osborne and Smith, 2005). In addition, the appropriateness of the culture media used and culture conditions will affect the fraction of this culturable community that is successfully recovered. For instance, many studies of fish gastrointestinal microbiota have not included anaerobic culture conditions and have therefore missed certain fastidious and obligate anaerobes completely (Nayak,... 

Viruses Typically, viruses are identified via culture-based methods and microscopy these methods are extremely slow with respect to the rate of infectivity of many viruses. Therefore, using a molecular-based technique with a rapid analysis time is imperative for biological defense applications where rapid identification of the microorganism is vital. The ability of PCR/ ESl-MS to characterize viruses allows for the immediate serotyping of the virus and identification of new strains. In addition, it may predict an emerging outbreak and therefore provide information to make decisions about quarantine, and allow medical personnel to provide appropriate supportive care. 

In industrial production quantitative analysis of the microbiological burden of the product is carried out before and after every micro-organism reducing step. Rapid microbiological methods (RMM), such as ATP-bioluminescence are capable of producing results in real time, since traditional culture-based microbiological analysis techniques take at least several days to get a result [38, 39], see also Sect. 19.6.5... 

The microbiological methods are listed (culture - traditional culture-based microbiological techniques, FISH - fluorescent in situ hybridization,... 

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