An Exercise, A Gram Stain Reaction

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In the first part of the practical exercise, a gram stain reaction was performed on the sample collected from the carrot infected with soft rot. Gram stain result showed that the bacteria presented on the diseased carrot were gram negative and rod shaped, which were stained pink under the microscope. This result indicated that the bacteria did not have a thick peptidoglycan layer; therefore it was not possible for them to retain the dark crystal violet stain, even with the presence of the mordant iodine solution. Violet crystals diffused out of the bacterial membrane and got washed away by alcohol, thus the membrane absorbed the counterstain carbol fuschin. The process of Gram stain gave rise to a few possible errors. If each step was not carried out properly or if the timing was not right, it could give rise to false results. When Gram stain was performed, all crystal violet dye could have possibly not all been washed out, as this had occurred to other groups, resulting in some bacteria to appear as purple under the microscope. On the other hand, during the decolourisation step, if there was too much alcohol added or if the alcohol was left on the slide for too long, all bacteria could lose their purple colour and start to take up the counterstain dye, resulting in a false gram negative result. Streak inoculation was performed on nutrient agar (NA) and MacConkey agar (MAC) plates. There are a few possible errors that could occur at this stage as well, due to improper application of aseptic methods. The agar plates could have been contaminated with microbes from other sources, and this could also affect the subsequent steps of the experiment. At this point, the first postulate was satisfied, as supported by the class data for Gram...

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...on cutting edge techniques gene identification and DNA microarrays, where the complete genomes of the disease causing microbes are determined, together with the complete genomes of different hosts that they infect (Cummings and Relman, 2000). These host-pathogen relationships are still yet to be determined, based on these modified Koch’s postulates and they promise many future successes for microbiologists to determine the aetiology of microbial diseases.

F. Conclusion
Without all of Koch’s postulates being satisfied, it is not adequate to conclude that E. carotovora is the only causative bacteria for soft rot in carrot. With all postulates validated except for the last, it can be concluded that due to a few suggested errors, another species is found that is likely to be similar to E.carotovora in its ability to cause soft rot, gram stain reaction and cell shape.

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