The results of the experiment supported the aim by identifying whether or not microorganisms found in dam water were disease- causing pathogens. The pathogens that were identified are opportunistic and can cause disease to the host by interrupting normal bodily functions.
Specimens may contain several species of organisms and therefore primary isolation allows the identification of specific bacteria to be accurate.7 By isolating the organism in nutrient agar, two separate colonies of bacteria were identified. Unknown microorganism one had a red pigment called prodigiosin produced by secondary metabolic activities of some microorganisms at temperatures lower then 37℃.8 This compared to microorganism two which had a cream/ light blue pigment called pyocyanin, a toxin produced and secreted by the organism clearly exhibits that two individual organisms were present.9 Due to the red pigment of microorganism one all bacteria except Pseudoalteromonas rubra, Serratia marcescens, Vibrio ruber, and Hahella chejuensis can be excluded.8 The light blue pigment on the second organism can exclude all bacteria except Pseudomonas aeruginosa.9
Using a differential stain then allows bacteria to be divided into two groups: Gram positive, which retain the crystal- violet dye due to a thick peptidoglycan layer and Gram negative which can be decolourised to allow a counterstain of safranin.10 This is due to a thin peptidoglycan layer and an additional outer membrane composed by phospholipids
The purpose of this lab was to identify two unknown bacteria from a mixed culture. The reason for identification of unknown bacteria was to help students recognize different bacteria through different biochemical tests and characteristics. This is important in the medical field because identification of unknown bacteria can help treat a patient by knowing the contributing source of a disease. Also knowledge of different bacteria helped others make antibiotics used today. This lab was completed by using the methods learned thus far in identification of bacteria.
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
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.
In a laboratory setting, it often becomes necessary to identify an unknown organism. In this experiment, researchers classified an unidentified bacterium based on its physical structure, colony morphology, optimal conditions and metabolic properties. A Gram stain using crystal violet, iodine, and safranin and a simple stain using methylene blue characterized the organism’s cell wall. Cultural behavior was classified by inoculating the organism onto nutrient agar and incubating it at 37° C for 48 hours, and observing its behavior, as well as using SIM medium to test for motility. Optimal growth temperature was
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
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)
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
The purpose of the following study is to determine where the two unknown bacteria acquired in Microbiology lab should be classified in regards to temperature, pH level, and osmoregularity. It is important to classify bacteria in order to identify them. Identification of bacteria is important because they are not only useful but potentially dangerous as well. The identification of bacteria can lead to breakthroughs in healthcare regarding treatment of old and new diseases alike. Identifying bacteria can also be used in many other areas from better crop production through microbial pesticides to biological warfare. Their uses are endless as are their abilities to evolve and adapt to changing environments. That is why it is so important
This experiment was given to us to utilized previous knowledge learned throughout the semester to identify a gram negative unknown bacterium. We had to first learn the difference between a gram negative and a gram positive organism. We started off with doing gram stains to determine whether it was positive or negative. Based on the gram stains, a gram positive stains purple and a gram negative stains pink. A gram positive stains purple because the cell walls is made of a thick peptidoglycan layer and doesn’t
As the flowchart shows, a series of tests were conducted to identify the unknown bacterium #65. Microscopic observation of the gram stain indicated a gram-positive coccus bacterium. S. epidermidis was used as the gram-positive control while E. coli was used as the gram-negative control. This observation led to the elimination of all gram negative and rod-shaped genera: Enterobacter, Citrobacter, Klebsiella, Escherichia, Pseudomonas, Serratia, Alcaligenes, Neisseria, Proteus, Salmonella, Shigella, Erwinia, Veillonella, Flavobacterium, Bacillus, Arthrobacter, Lactobacillus, Listeria and Kurthia (2). By performing the catalase test, it was determined that the bacterium was catalase negative and it did not produce bubbles. M. luteus and E. faecalis were used as positive and negative controls, respectively.
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
In the world of microbiology it is vitally important to be able to discern the identities of microorganisms. Not only is it important in a lab setting but as well as in healthcare in general. Properly identify what strain of bacteria a person has will aid in the proper medicine and dose given. Throughout the semester we have learned about different types of bacteria and certain test that can clearly identify them. The purpose of this lab report is to identify a Gram-positive or Gram-negative bacterium. Using all the knowledge of procedures and lab techniques identify the unknown and discuss all the tests you performed.
Escherichia is a genus of aerobic gram-negative rod-shaped bacteria of the family Enterobacteriaceae that form acid and gas on many carbohydrates, such as dextrose and lactose, but not acetone, which include occasional pathogenic forms, including some strains of E. coli which are normally present in the human intestine as well as other forms which typically occur in soil and water (Webster). Escherichia coli is a gram-negative bacilli that rarely varies in shape and size and when stained often resemble safety pins because the ends of some bacilli stain more densely than does the middle; which is a characteristic called bipolar staining which is common in enteric gram-negative bacilli (ASM). Gram negative cells have a thin cell wall layer and will stain red to pink. The staining process is the same as Gram positive, requiring four steps: applying a primary stain, adding a mordant, then rapid decolorization and completing with a counter stain. Applying the alcohol for decolorization dissolves the outer membrane and leaves small holes in the thin peptidoglycan layer through which the crystal violet-iodine diffuse. The gram-negative bacteria is colorless after the decolorization; therefore adding safranin
The bacteria that was contained within Unknown tube #12 is believed to be Pseudomonas aeruginosa, Figure 1. The bacteria tested to be Gram Stain negative, producing a pink, red color retained from the staining process. When the species of bacteria was plated on nutrient media, the cells produced an irregular and spreading configuration as shown in Figure 2. This same plating test provided the margins and elevation, lobate and hilly, respectively. The specimen was stabbed in a Fluid Thioglycollate Medium (FTM) tube using an inoculated loop of the bacteria. The results of this experimentation indicate the type of oxygen requirement of the bacteria. The test found the bacteria to be aerobic as colonies of the bacteria began to form along the top of the FTM tube (Manual 2017).
Introduction: Through the conduction of numerous experiments, the identity of two bacterial isolates was determined. The tested specimen was an unknown sample of a mixed culture of two different species of bacteria. The first step that was taken was obtaining a pure culture of each species of bacteria by isolating one species from the other. Once isolation was complete, the isolated cultures were tested using procedures that had been performed during previous lab sessions. A gram stain was performed on the two isolates. The isolate which had tested gram negative was then tested for the presence of cytochrome C and lactose fermentation. For the gram positive isolate, cell shape was determined and a catalase test was performed.