FISH analysis showed that the new extraction method was successful for four out of seven probes used on the sludge samples. The general probe EUB 338 showed a high density of bacteria in the seeding sludge and highly reduced densities after 24 and 28 days of operation, though still some patches with high bacterial density could be detected. In the samples from days 24 and 28 cloudlike structures were observed in all wells including negative controls, which showed high light intensity. These structures coincided with reduced hybridization ability of the EUB 338 probe (cf. figure 14 G-L) Whether the background of these structures was too high, the probe directly bound to the substances, which caused these structures or the hybridizing or fluorescent ability of the …show more content…
With NIT 3 probes hybridized bacteria could not be detected, thus either hybridization was unsuccessful or nitrate reducing activity by the NIT 3 gene was not present in the samples. Showing fewer artifacts, the Ntspa 662 probe showed no positive hybridization. Ntspa 662 is specific for the genus Nitrospira, which is known to conduct nitrite oxidation. For the BS 820 probe, which is specific for Candidatus Scalindua wagneri and C. S. sorokinii, no hybridization to bacteria was found, while for the more general probe Sca 1309 just one very small colony of Candidatus Scalindua specimen could be detected (figure 15, J - L) and a very high abundance of anammox bacteria of the genera Kuenia stuttgardiensis or Brocadia anammoxidans was verified using AMX 820 probe, especially in the seeding sludge samples. Thus the majority of anammox bacteria in the seeding sludge are not of the genus Candidatus Scalindua. Besides the positive FISH results, the shape of bacteria hybridized with AMX 820 was similar to what was initially seen with DAPI. Fluorescence reveals ring and sickle shapes (cf. figure
The following tests according to the lab manual were performed: gram stain, fermentation tubes, methyl red, vogues proskauer, sulfur, indole, motility and growing it up on MacConkey agar. The gram stain was performed incorrectly the first time. This is because the decolorizer was not on the bacterium slide for long enough, giving a false outcome.
There are many differents ways to identify a bacterial unknown and many different situations where identification would be beneficial. One way to identify bacterial unknowns is to perform biochemical tests. In this experiment multiple biochemical tests were done, by performing these tests on the bacterial unknown received the two different bacteria were then identified. The citrate test is done to test the ability of organisms to use citrate as a carbon source. This test uses Simmons citrate agar, the agar contains sodium citrate as the only carbon source and has bromothymol blue as the pH indicator. The organisms that use citrate as a carbon source use the enzyme to transport the citrate into the cell. The cells converts ammonium dihydrogen
An unknown bacterium was handed out by the lab instructor. The methods that have been learned so far in identifying bacteria were applied to this unknown. Procedures were followed as stated in the lab manual and biochemical test handouts. The first procedure that was done was a gram stain followed by a streak of the unknown on a TSA plate in order to determine the gram reaction and observe the colony morphology. After that, specific biochemical tests were performed for gram positive, since unknown number five was determined to be gram positive rod. The other tests were performed in this order: Mannitol Salt (MSA) streak, Blood Agar streak, Catalase test, Nitrate Reduction test, and Phenyl
After the incubation period the bacteria was observed for pure colonies. The colonies were sampled and the three streak plate technique was repeated and this sample was incubated for forty eight hours at 37 degrees Celsius. After the incubation of the colonies, a gram stain was performed which is defined in the lab manual.
The first result of importance was the result of the Gram stain. The observations of the unknown bacteria from the slant culture after Gram staining showed that the unknown bacteria were Gram negative bacilli (Image 1). After determining the unknown bacteria was Gram negative, an oxidase test was conducted on a sample from the slant culture. The cotton swap with the sample of bacteria did not change color when the oxidase reagent was applied, thus providing a negative result. With a negative oxidase test, further tests were conducted to determine various characteristics of the unknown bacteria. A MR-VP broth was inoculated with a sample from a slant culture of unknown bacteria. After incubation, the methyl red reagent was added to the broth, and the broth turned red, providing a positive result (Image 2). An EMB agar streak plate was inoculated with a sample from a slant culture of the unknown bacteria, and after incubation, growth was found on the plate, providing a positive result (Image 3). A Citrate agar slant was inoculated, and after incubation, growth was found on the media, providing a positive result (Image 4). A Urea agar slant was inoculated, and after incubation, the agar had changed from a peach color to a bright pink color, providing a positive result (Image 5). Using the flowchart (Figure 1) developed from the Table of Expected Results, the lab partners started at the oxidase test. Given the negative result of the oxidase test, the flowchart is
Today in medicine doctors are rapidly isolating and distinguishing the many pathogenic microbes encountered daily within the environment. Public health has been affected from the faster identification of microorganisms by delivering an accurate analysis to patients in order to receive treatment of the disease in a timely manner. Due to the growing understanding of these organisms more have been easier to indicate to improve water quality. Also more methods have been developed for better treatment options from fecal bacteria in public water systems. Scientist has developed such specific methods of identifying the unknown organism to tell if the contamination has come from either a human, bird, or mammal. (Achtman et al., 2008)
The purpose of this lab was to identify two unknown bacteria cultures using various differential tests. The identification of these unknown cultures was accomplished by separating and differentiating possible
The purpose of this study project was to carefully isolate and identify two unknown bacteria from a mixed culture. The ability to properly evaluate biochemical test results is also necessary for the identification to be successful. The goal was to apply all of the methods and techniques that have been learned in the microbiology laboratory course for the proper identification of unknown bacteria. A certain amount of bacteria that were used throughout the course were possible bacteria that could be found in a mixed culture. The bacteria that were identified in the mixed culture were Staphylococcus Aureus and Kocuria Rhizophila.
The purpose of this study was to identify the unknown bacterium using biochemical tests and various methods that had been learned from previous the microbiology laboratory class. Identifying the unknown bacterium was determined by separating and differentiating possible
The purpose of this experiment is to obtain isolation of individual species of particles from the mixed culture. This is completed through the isolation technique of streak plate. The objective of this experiment is to replicate the technique of streak plate but on a much larger scale. Because it is on a larger scale the particles are able to be visually observed as they are isolated using the streaking technique as the experiment is conducted. The benefits of the streaking technique is when a cultures has multiple species they are able to be more easily identified once they have been isolated. This experiment is much like the experiments completed on an agar plate but on one a much larger scale and where techniques
This laboratory experiment’s objective was to take a pure culture and isolate it from a mixed culture. The other part of the objective was to ascertain what species of bacteria that the pure culture was. The hypothesis made stated that so long as lab protocol was followed, the unidentified culture would be positively recognized/identified. An isolated pure colony of the unknown culture was obtained using the quadrant streak plate method. Afterward, the culture was Gram stained, and the results showed that it was Gram positive. Motility tests were done on the unknown using a filter paper bridge on a petri dish that contained TTC with agar. The unknown was revealed to not be motile, which meant that it did not possess flagella. The last test done was to learn the metabolic capabilities of the unknown bacteria. There were tests done for citrate utilization, the mixed fermentation pathway, catalase presence, carbohydrate fermentation in mannitol, lactose and glucose, urease production and the butanediol fermentation pathway in order to better identify the unknown bacteria. The results from each of the metabolic tests in conjunction with the motility and Gram staining tests were ultimately compared to results from database containing many different kinds of results from various bacteria. The unknown from the mixed culture was identified as Staphylococcus
See Table 1 and Flow Chart 1 for results of Bacteria # 1 and Table 2 and Flow Chart 2 for results of Bacteria # 2.
In order to obtain well-isolated discrete colonies, the quadrant streak and spread technique was used. This allowed dilution of the original microbial material over the entire surface of the plate. As the original sample was diluted by streaking and spreading it, over successive quadrants the number of organism decreases. Usually by the third or fourth quadrant only a few organism were transferred on the by the inoculating loop and theses produce a few isolated
In suspended growth processes, the microorganisms responsible for the treatment of wastewater are maintained in liquid suspension by appropriate mixing methods. Many suspended growth process
Each mixed culture that was tested had one gram positive and one gram negative bacterial species. The possible species of bacteria that could have been isolated from the mixtures included the following: Bacillus subtilis, Corynebacterium glutamicum, Escherichia coli, Staphylococcus aureus, Enterococcus faecalis, Enterobacter aerogenes, Salmonella enterica, and Pseudomonas aeruginosa. The identities of the unknown species were determined through comparing the experimental data against data acquired from earlier experimentation.