Inoculation Technique
Culturing of organisms has become a global practice in business and agriculture. The approach has been depicted to help improve the population and quality of breeds of various microorganisms. In botany, the process has aided in improving plant quality and sustainability. Most trees, especially fruit trees, have been enhanced to enhance fruit quality (Sanders, 2012, pg. 6). However, most plants with unwanted characteristics have been controlled to become more desirable in bearing. In zoology, the practice entails the provision of a conducive environment for propagation. Breeding has become accessible through the application of the various techniques of cultivation. This paper seeks to evaluate, analyze, compare and explain the methods used to cultivate microorganisms. It demonstrates the skillful applications of aseptic techniques by carrying out practical investigations into the growth of microorganisms.
There are different techniques used for culturing the organism. The choice of the particular mechanism depends on suitability and the growing requirement of the organism (Overmann, Abt, & Sikorski, 2017. pp.711-730). These techniques include; inoculation, incubation, isolation, inspection, and identification.
The inoculation technique involves the introduction of organisms to a conducive environment that will enhance their growth and reproduction (Overmann, Abt, & Sikorski, 2017. pp.711-730). This practice entails introducing some substances to the environment provided with suitable nutrients needed by the organism. The technique involves five media types: Agar Plates, Broth Culture, Slant culture, Plate culture, and Stab culture (Phelan & May 2015, pg.66-120).
Agar plates form the traditional media utilized for bacteria growth. Nutrients are mixed with agar, thus providing a conducive environment for bacteria growth. The solution is inoculated on plates through streaking. A steaking loop is dipped in the solution with bacteria cells streaked on the plate culture (Phelan & May 2015 pg.66-120). The plates are then stored at suitable temperatures and expected to aid in the performance of further studies on the organisms. The method can also involve using a liquid media where a single culture of the bacterial microorganisms is added to a small quantity solution forming one mixture, and then pipetted in the liquid media culture (Phelan & May 2015, pg.66-120). The availability of nutrients and other compounds in the mixture is essential for the growth of the microorganism. However, the mix of the liquid media and the bacterial cells have to be placed in a conducive environment, especially at room temperature, for growth.
An inoculation indicator is used to inoculate the antiseptic broth culture in this method. Glowing the exposed end of this broth shall help to preserve it sterile. The broth is stirred up to the pointer; thus, the stick’s tip is immersed while keeping the novel state of this needle.
The twirling of the pointer carefully aids the inoculation of these bacteria from the stick towards the antiseptic broth (Sanders, 2012, pg. 300). An inoculated soup culture can then be detached from the pointer. The rod is kept sterile using an aseptic method that prevents contamination. The methods can also involve using a soothing liquid that is prepared by using a plethora in a tube (Phelan & May 2015 pg.66-120). The broth is made by mixing yeast, sodium, tryptone, and chloride.
Agar Plates
An inoculation needle is used to achieve the fishtail inoculation method in the slant culture method (Stacey, 2011pg. 200). Microorganisms are removed from their original environment to the inoculation needle after uncapping the sterile slant. The uncapped end of the slant culture has thereafter flamed the state of the plant is maintained and moves the needle upwards until the inoculation needle’s tip contacts the bottom of the sterile media. A windy pattern is portrayed on the agar surface after the needle inoculates the agar through the use of the media an aseptic method is used in the removal of the needle (Phelan & May, 2015, pg.66-120).
The method entails using a flashing technique in the formation of the steak dish. The inoculation needle flashed across the dish in some determined directions after the lid is lifted to hover on top of the agar dish (Stacey, 2011pg. 200). There is the creation of microbial vaporizers done as a result of striking the inoculation tip on the sides of the agar dish. The inoculation needle is then separated from inoculated agar dish culture and flamed.
An inoculation needle is a vital tool in the stab culture. The secure stab culture lid is done, and the unclosed end of the stick is flamed. The tip of the needle is forced into the stab media till it reaches 0.5 inches from the foot of the stab media (Overmann, Abt, & Sikorski, 2017. pp.711-730). The needle is separated from the media in a similar direction, thus preventing a wobbling effect that can affect the culture. The flame is used to sterilize the needle.
Unlike the other microorganism culture methods, inoculation involves mixing the bacterial cells with the inoculated solutions.
The process entails providing a conducive hutching condition to the microorganisms (Talaro, Chess, Wiersema & Sen, 2013, pg. 687). The method involves using an insulated and closed device that gives ideal humidity, temperatures, and other environmental conditions for the growth of organisms. The incubation provides a temperature of 37°C and is kept for between 12-18 hrs. The bacterial cells are placed in an incubator with conducive conditions for the growth of the organisms (Overmann, Abt, & Sikorski, 2017. pp.711-730).
The method involves the removal of the strains from a natural, mixed population of microorganisms in the environment (Talaro, Chess, Wiersema & Sen, 2013 pg. 687-704). The method aims at separating some pure breeds of microorganisms from the rest at helping them to grow and reproduce. The process involves techniques like;
Streaking; entails pouring an appropriate sterile media into a Petri dish and allowing the media to solidify. A sterile needle is used to streak a small growth quantity from the chosen broth culture across, back and forth, the agar surface till dish’s diameter is covered (Talaro, Chess, Wiersema & Sen, 2013 pg. 687). The needle is then flamed while streaking is continued at the right angles and across the first streak. The process helps to drag the bacteria forming a line from the first streak. The needle flamed after the streaking and continued on the second streak, then parallel to the initial one.
Broth Culture
Planting: diluting a mixture of the microorganism until a few bacteria are left in milliliter suspension. A small amount of the dilute is put in a sterile petri-dish using a sterile pipette. Melted agar is then cooled to 45 °C then and poured onto a dish. The agar and the microorganism are well mixed. After the solidification of the agar, the bacterium shall be collected and become visible and grow into a colony (Talaro, Chess, Wiersema & Sen, 2013 pg. 687).
Dilution: the method is helpful for those microorganisms that cannot be separated by streaking and planting techniques (Talaro, Chess, Wiersema & Sen, 2013, pg. 236). The method targets separating the microorganisms that are outstanding from the rest. The process involves transferring a mixture from one sterile tube to the other until the microorganisms are obtained.
Enrichment method: this used the media and other cultivation conditions that promote the growth of some desired organisms. The process entails providing a conducive environment for certain microorganisms while functioning to eliminate others (Vartoukian, Palmer & Wade, 2010, pg. 345).
This entails observing the usual growth properties that are useful for the analysis of the specimen elements. The method involves collecting samples and detecting the availability of microbes via culturing method. The observation is made on the suitable condition for the growth of the microorganism and providing the suitable environment for their development.
This techis the widely used method of molecular identification. It involves PCR, the most reliable method of identifying microorganisms (Talaro, Chess, Wiersema & Sen, 2013, pg. 236). It deals with the spectrum of tests applicable in various forms of cell detection and the identification of most tinny organisms.
Conclusion
In all the various techniques of culturing the microorganism, the choice depends on the suitability of the method to collect the desirable size and type of the organism. The provision of a favorable conditions environment is vital for the survival of the microorganisms. These methods differ in the forms of technology utilized and the effectiveness of collecting enough samples and propagating a higher number of microorganisms. The importance of microorganism culturing is to aid in reproducing the organisms to provide enough specimens for research. Learning about microorganisms is vital because it aids in the identification of the best methods to control the spread of pathogens and germs that are found in food matter.
Reference List
Overmann, J., Abt, B. and Sikorski, J., 2017. Present and future of culturing bacteria. Annual review of microbiology, 71, pp.711-730.
Phelan, K. and May, K.M., 2015. Basic techniques in mammalian cell tissue culture. Current protocols in cell biology, 66(1), pp.1-1.
Sanders, E.R., 2012. Aseptic laboratory techniques: plating methods. JoVE (Journal of Visualized Experiments), (63), p.e3064.
Stacey, G.N., 2011. Cell culture contamination. In Cancer Cell Culture (pp. 79-91). Humana Press.
Su, C., Lei, L., Duan, Y., Zhang, K.Q., and Yang, J., 2012. Culture-independent methods for studying environmental microorganisms: methods, application, and perspective. Applied microbiology and biotechnology, 93(3), pp.993-1003.
Talaro, K., Chess, B., Wiersema, D.S. and Sen, P., 2013. Foundations in Microbiology, 2012. McGraw-Hill.
Vartoukian, S.R., Palmer, R.M. and Wade, W.G., 2010. Strategies for culture of ‘unculturable bacteria. FEMS microbiology letters, 309(1), pp.1-7.
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