Mediatube stock2/2/2024 ![]() ![]() In these situations tubes or flasks which can be tightly sealed with a butyl rubber stopper are used. However, it often is desirable to grow cultures outside of the chamber due to limited space or the need to frequently monitor growth. For experiments in which the culture will remain in the anaerobic chamber or jar, any type of closure that maintains sterility will suffice. Culturing performed entirely in the glove box will result in more consistent growth patterns with shorter lag periods which are necessary for some physiological studies but may be of no consequence when simply inoculating a series of overnight cultures for DNA isolation.Īnother important consideration when growing cultures in broth is the type of tube or growth vessel to be used. The choice of whether to work on the bench top or in an anaerobic chamber/glove box depends on the needs of the experiment. See UNIT 1A.1 and other pertinent resources ( APPENDIX 1B) for more information.īacteroides cultures must be incubated anaerobically to allow growth, but since they are aerotolerant many manipulations can be done conveniently on the bench top. However, given that some procedures described in this unit require the use of syringes for the transfer of cells, it is recommended that the standards for BSL-2 be followed. These organisms are not known to consistently cause disease in healthy adult humans, and are of minimal potential hazard to laboratory personnel and the environment. However, this unit is not intended to be a comprehensive treatise on anaerobic culturing methods and the reader will be directed to several excellent reviews and manuals for detailed descriptions of anaerobic techniques.ĬAUTION: Bacteroides are considered Biosafety Level 1 (BSL-1) organisms in the USA although in some European countries the are BSL-2. It is our intention to describe sufficient equipment options so that those new to the field will be able to easily initiate Bacteroides culturing without a large initial investment in expensive equipment. ![]() In this regard, it is important to note that anaerobic culturing requires some specialized equipment. ![]() Thus, while it is appropriate that this unit include a discussion of anaerobic culturing techniques, the methods discussed here may not be suitable for many other anaerobic bacteria since they are not as rigorous in their exclusion of oxygen at all stages of media preparation and inoculation. That said, they are one of the most aerotolerant anaerobic species known and a recent report ( Baughn and Malamy, 2004) has suggested that they may actually slowly divide in the presence of nanomolar concentrations of oxygen. The single most important consideration when culturing the Bacteroides is that they are obligate anaerobes and they will not divide in the presence of oxygen. Although there has been considerable interest in the mechanisms responsible for the transmissible, acquired antibiotic resistance, it is their innate resistance to aminoglycosides and vancomycin that has proven to be important for development of selective media for their isolation (see Smith et al., 2006). Due to a combination of innate and acquired mechanisms the Bacteroides species are resistant to a wide array of commonly used antibiotics which can complicate antimicrobial therapy of anaerobic infections. All of these factors are important considerations when culturing these organisms. Their ability to successfully colonize mammals is due in part to their extended aerotolerance, simple nutritional requirements, and ability to utilize diverse carbohydrate substrates for carbon and energy. Largely due to their proximity to the human host, several Bacteroides species can be opportunistic pathogens and they are frequently isolated from a range of anaerobic infections including intraabdominal and pelvic abscesses, soft tissue, female genital tract, and bacteremia. They are the predominate members of the indigenous intestinal flora in humans where they contribute to normal intestinal development, physiology and function. This unit describes methods for culturing, storage, and maintenance of the Gram-negative, anaerobic Bacteroides species Members of the genus Bacteroides play important roles in human and animal health ( Smith et al.,2006).
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