Aseptic transfer

Based on the above discussion we are now ready to start a real experiment. Molasses are transferred from the sugar industry and kept in a cool room. The ATCC culture order has arrived and hydrated. Stock culture was prepared and aseptic transfer successfully done. 

Biological indicators (Bis) for use in thermal, chemical or radiation sterilization processes consist of standardized bacterial spore preparations which are usually in the form either of suspensions in water or culture medium or of spores dried on paper, aluminium or plastic carriers. As with chentical indicators, they are usually placed in dummy packs located at strategic sites in the sterilizer. Alternatively, for gaseous sterihzation these may also be placed within a tubular hehx (Line-Pickerill) device. After the sterilization process, the aqueous suspensions or spores on carriers are aseptically transferred to an appropriate nutrient medium which is then incubated and periodically examined for signs of growth. Spores of Bacillus stearothermophilus in sealed ampoules of cultrrre medium are used for steam sterilization morritoring, and these may be incubated directly at 55°C this eliminates the need for an aseptic transfer. 

Aseptic transfers are also avoided by the use of self-contained units where the spore strip and nutrient medium are present in the same device ready for mixing after use. 

Appropriate Xenorhabdus or Photorhabdus bacterial cells are aseptically transferred to 5 ml of nutrient broth in a test tube and kept overnight on a shaker. The flasks containing autoclaved material are inoculated with the bacterial culture by pouring the contents of one culture tube. The flask is shaken well and stored for 2-3 days at 25°C to allow multiplication of the bacteria. 

Inoculation and Aseptic Transfer Monitoring of Growth Parameters and Control Containment and Containment Control Ascites Production Mouse Colony... 

A culture of M. isabellina CCT3498 was aseptically transferred into conical Erlenmeyer flasks (500 mL) containing 200 mL of sterile PDCB and kept on a rotary shaker (150 rpm) at 30 °C for 3 days to acquire biomass. 

After the cycle, aseptically transfer the spore strip to vessels of culture media. If spore suspensions were used, aseptically transfer the inoculated bottles to a laminar airflow workstation and add culture media to the bottles. Use appropriate positive and negative controls. 

Following the dry-heat cycle, aseptically transfer the units containing endotoxin to an aseptic area for extraction procedures, sampling, and conducting the limulus amebocyte lysate (LAL) test. 

The contents of the flasks are pooled, a 25 ml inoculum (taken from the pool) is added to each of twenty 2-liter flasks, each containing 500 ml of the following sterile medium 30 grams soybean meal 40 grams dextrose (cerelose) 1 gram calcium carbonate 1,000 milliliters tap water. The flasks and their contents are incubated for 3 to 5 days at 28°C on a rotary shaker (280 rpm, 2 inch stroke). The broth is pooled and aseptically transferred into a sterile inoculum flask having a side arm (total volume, about 10 liters). 

Germination stage 2 Aseptically transfer 25 ml of the fermentation medium of Germination stage 1 to a 2-1 shake flask containing 500 ml of the above described sterile germination medium. Incubate the flask and its contents for 3 days at 28 ]C on a rotary shaker (280 rpm, 2" stroke). 

Fermentation stage Aseptically transfer 500 ml of the medium obtained from Germination stage 2 to a 14-1 fermentation tank containing 9.5 I of the following sterile medium ... 

Many of these preventive measures are made much easier if, instead of a small room, a cabinet is used to carry out aseptic transfer (see below). 

All bottles of medium prewarmed by standing in a water bath should be thoroughly dried with tissues (not a dirty towel) before being taken into the aseptic transfer room. Water baths should contain bacteriostatic agents, e.g. Panacide (British Drug Houses Ltd. Appendix 3). 

Aseptic processing in the pharmaceutical industry is almost entirely dependent upon the proficiency of the personnel assigned to this most critical of all activities. The operators must be able to consistently aseptically transfer sterile equipment and materials in a manner that avoids contamination of those materials [1]. This is no mean feat given the contamination continuously released by personnel and the prevailing need for personnel for execution of the process activities. 

The USP describes two general methods for conducting the test the direct transfer, or direct inoculation, method and the membrane filtration method. As the name indicates, the direct inoculation method involves the aseptic transfer of a sample of test product solution into the sterility test growth medium. To use this method, it must first be demonstrated that the product solution itself does not inhibit the growth of typical indicator microorganisms specified in the USP method. It should be self-evident why it is important to perform testing to negate the chance of product inhibition of possible microbial contaminants, as this is the purpose of the sterility test. The direct inoculation method, while not theoretically complex, requires the utmost technical precision and aseptic manipulation techniques for proper execution. As a consequence of the repetitive motions involved, it is prone to human error. 

A suitably sized solution preparation system similar to that mentioned under the previous sections can be used to provide material for bulk freeze drying. (Since product solutions can be sterile-filtered directly into the final container, microbial and particulate exposure will be minimized.) The sterile solution is subdivided into trays and placed into a sterilized freeze dryer. Aseptic transfer of sterile product in trays to the freeze dryer must be validated. After tray drying, the sterile product is aseptically transferred through a mill into suitably designed sterile containers. The preparation of sterile bulk material is usually reserved for those cases where the product cannot be isolated by more common and relatively less expensive crystallization methods. Due to recent advances in this field, a freeze drying process should be considered as a viable option. ... 

Powder fills are made by aseptically transferring the sterile bulk powder from its containers into the hopper of the filling machine. The transfer is usually done from a container that is mechanically positioned over the hopper with a solid aseptic connection to the hopper. 

B. Shake flask. A 2800-mL Fernbach flask fitted with a vapor bulb and an air inlet (see Figure 1) containing 500 mL of MSB solution with 0.2% L-arginine-HCI is sterilized (Note 2). After cooling, the 50-mL preculture is added via aseptic transfer (Note 7). The vapor bulb is filled with 10 mL of chlorobenzene, and the flask is carefully placed in an orbital incubator shaker at 150 rpm and 30°C for 48 hr. The excess chlorobenzene is removed from the central chamber with a pipette, and the pH... 

An aseptic transfer involves first passing the lip of the preculture flask through a flame and repeating the flaming procedure with the lip of the Fernbach flask. 

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