Francisella tularensis Imagine waking up one morning with inflamed eyes, a sore throat and severe headaches. Within a few days time you start having trouble breathing and develop pneumonia. With no idea of what’s wrong and no treatment in sight, you may die anywhere within a 5-14 day range (10). At the dawn of the 20th century, this type of scenario was not uncommon. By the early 1950’s the condition plagued thousands of people each year. It wasn 't until 1959 when the condition actually had a name...tularemia (16).
Tularemia is a disease caused by the bacterium Francisella tularensis. F. tularensis is a microorganism belonging to the Domain Bacteria, Phylum Proteobacteria, Class Gammaproteobacteria, Order Thiotrichales, Family Francisellaceae, Genus Francisella and species tularensis. This microorganism is considered to be in the Domain Bacteria rather than in the Domain Archaea for a few reasons. Not only does F. tularensis not have a nucleus, but it is also unicellular and contains a single circular dsDNA. The plasma membrane is also composed of a series of unbranched fatty acids that are connected to glycerol molecules via ester linkages, all characteristic of Domain Bacteria (5).
Characteristics of F. tularensis’ next taxonomic rank, as part of the Phylum Proteobacteria, characterize it as a Gram-negative bacterium. When gram-stained, this microorganism appears as a reddish color under the microscope. This is mainly because Gram-negative bacteria have an inner
In this experiment, an unknown bacterium was given to each individual student. The main purpose of this lab was to identify the given unknown bacteria going through a series of biochemical tests as one of the gram negative bacteria among six different Gram negative bacteria Escherichia coli, Enterobacter aerogenes, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa and Salmonella typhimurium. At the very beginning, streaking method; T-streak technique was used to isolate the pure colonies. For the morphological identification of unknown bacteria, Gram Stain Method was done. Biochemical tests that were conducted for the experiment
The purpose of this study was to determine what an unknown bacteria was using several different microbiology lab techniques including an API test, an oxidase test, a gram stain, a hanging drop slide, and morphology identification. The unknown bacterium, which was contaminated with Serratia marcescens, was isolated by streaking the bacteria solution to single colonies. The isolated unknown white bacteria, had the appearance of circular form, convex elevation, entire margin, elongated cocci. The tests than showed that the bacteria was gram-negative, non-motile,
The next step of the project included preparing a Gram stain to discover the cell shape, arrangement, and if the bacteria is gram positive or
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 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).
Thus we inoculated lactose broth, glucose broth and tryptophone broth with our organism. At our final lab meeting, we observed that the lactose broth was still phenol red in color, glucose tube had turned yellow with a bubble present in Durham tube, and the oily ring in our Indole test was colorless. Taking all the results into consideration and rechecking all of our steps against the guidelines and steps to ensure all required tests had been performed, we concluded that our organism belonged to Group IX genre, Proteus, Providencia, and Morganella. We then proceed to consult with our instructor.
Table 3 shows Gram stain results that indicated C. Freundii as a gram negative bacterium in rod shapes scattered in singles and some in pairs. Each gram stain produced the same results. The Bartholomew and Mittwer method of endospore staining indicated that C. Freundii tested negative for endospore formation. Table 4 shows the biochemical test results of the unknown and the official test results for comparison.
They are non-spore forming bacteria that are nonmotile and aerobic. F. tularensis grow best at thirty-five to thirty-seven degrees Celsius in the lab setting, while in nature, it can survive up to weeks on end at low temperatures. Fortier states that F. tularensis also contains siderophores, which are molecules that have the ability to bind to iron, that grow under iron-limiting conditions. This feature is crucial to the bacterium “because intracellular replication of F. tularensis is iron-dependent, as shown with deferroxamine having inhibitory effects in a tissue during infection” (Fortier et
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
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 colonies were smooth, translucent, and had a white brownish color. The Gram stain resulted in Gram positive cocci. After the Gram stain was completed, the bacteria were streaked on a Mannitol-Salt Agar plate and a Catalase test was performed. After these test were completed a Phenol Red Dextrose Fermentation tube was inoculated, and a SIM Tube inoculated.
Tularemia, otherwise known as Rabbit Fever, is a dangerous infectious disease of humans and animals that can cause severe complications and can be potentially fatal. Tularemia is commonly called Rabbit Fever because rabbits are known to be susceptible vectors. Other rodents and animals can become infected if they come in contact with the illness, including livestock and common house pets, such as dogs and cats. Tularemia normally occurs in heavily wooded, rural areas inside the United States.
In all areas of biology, it is easy to see that structure is related to function. This statement holds true in microbiology as well, the study of microorganisms, including bacteria. One characterizing feature of bacteria is the cell wall, which can generally (although not in all situations) be categorized into one of two categories: either Gram positive or Gram negative. Gram positive bacteria’s cell walls are composed of a large peptidoglycan layer (up to 90% of their cell wall). Within this large peptidoglycan layer, one can find techoic acids, which contribute to the maintenance of cell wall structure, and lipotechoic acids, which attach to membrane lipids. Gram positive bacteria that act as pathogens can also potentially release exotoxins, which can have very dangerous effects on humans. Gram negative bacteria, on the other hand, have a very small layer of peptidoglycan in their cell wall, which is surrounded by an outer membrane. Within the outer membrane, one can find the lipopolysaccharide layer, which is one of the most distinguishing factors of Gram-negative bacteria. It is important to note that Gram negative bacteria fail to possess techoic
M. tuberculosis cell wall is made of many lipids including peptidoglycan and mycolic acids. Hydrophobic molecules making up Mycolic acids form a lipid layer surrounding the organism affecting permeability of the cell surface. This layer is responsible for many pathogenic inducing traits of M. tuberculosis by inhibiting attack from lysosomes, reactive oxygen species (ROS) and cationic proteins in phagocytes. Further pathogenesis of the bacterium is achieved by the presence of other lipids which include cord factor, sulfolipids and phosphatidylinositol
These are gram positive bacteria which are found in acidic condition of pH 4 to 4.5 – acidophilic.