Hemolytic Activity Test / Sheep's Blood Agar Lab Report

The disease I chose to do my first paper on is Hemolytic Uremic Syndrome. This was a disease that I actually acquired as a young child and was diagnosed and treated for. The way it is described is a early destruction of red blood cells which affects the Cardiovascular system. It ends up clogging the filter systems of the kidneys and then at some point could lead to kidney failure if not treated correctly.

1. Hemoglobin is the molecule that carries oxygenated blood from the lungs throughout the body through inhalation and deoxygenated blood from the body to the lungs through exhalation. When the hemoglobin is in the oxygenated state, it is in the R state or relaxed state. When it is in the deoxygenated state, it is in the T state or tense state.

The patient is usually awake during angioplasty, and chest discomfort may be experienced during the procedure. The patient remains awake in order to monitor the patient symptoms. If symptoms indicate the procedure is causing ischemia the cardiologist may alter or abort the part of the procedure. Bleeding from the insertion point in the groin (femoral artery) and wrist (radial artery) is common, in part due to the use of antiplatelet drugs. Some bruising is therefore to be expected. But occasionally a hematoma may form. This may delay hospital discharge as flow from the artery into the hematoma may continue which requires surgical repair. Infection at the skin puncture site is rare and dissection of the access blood vessel is

Anemia conditions can be mild or severe, and the Social Security Administration looks closely at the severity of medical symptoms that negatively impact red blood cells. Some anemia conditions are so crippling that hospitalization and special medical treatment is the only way to get relief. Hemolytic anemia falls into this category. If you’ve been diagnosed with hemolytic anemia, and your condition is so acute that it makes holding down a job difficult, or impossible, you may be eligible for SSDI benefits.

Haemoglobin determination, or haemoglobinometry, is the measurement of the concentration of haemoglobin in the blood. Haemoglobin's main function in the body is to carry oxygen from the lungs to the tissues and to assist in transporting carbon dioxide from the tissues to the lungs. The formation of haemoglobin takes place in the developing red cells located in bone marrow. Haemoglobin values are affected by age, sex, pregnancy, disease, and altitude.

Isolation, gram staining and identification of the micro-organism is a basic diagnostic tool in clinical as well as in research. This method is useful in diagnosis and treatment of diseases. The gram staining is simple, least expensive and is rapid. It is useful in counting of total bacteria. Because of the staining, the gram positive and negative bacteria can be viewed under the microscope. The shape, size can be determined. The gram positive bacteria may lose the stain easily. The retention of stain depends on the age of the cell. Old cultures get readily decolorized. Yet, this method is feasible method for diagnosis of the minute

Through proper sterile technique and inoculation, I was able to capture maximum success in plate streaking for selective or differential medias and isolation of bacterium. In order for this lab to be an overall success the sterile technique, inoculation, and plate streaking were to be done with patience making it the much harder task in this lab. Observing the final plates for differential or selective, as well as for isolation were quite difficult to figure out as well. The media that I had more trouble in differentiating through the variety of characteristics and morphology was Sheep Blood Agar (SBA). Figuring out specific colonies was difficult due to beta hemolysis which is seen as a zone of clearing around the colonies. The easiest to distinguish of all three medias was obviously the Tryptic soy agar (TSA). Being the general growth media, we knew what we were expecting which was to see all three bacteria to grow thus making the media not selective nor differential. We knew in the case of Mannitol Salt Agar (MSA) that it could be selective or differential. Through the experiment we were able to notice which bacteria were salt tolerant and those that weren’t. In conclusion it was easily noticeable that S. aureus was able to ferment mannitol being very salt tolerant while the other bacteria struggled to prosper in

Mannitol Salt Agar is used to differentiate Staphylococcus areus from other microorganisms. The agar contains mannitol, salt, and a red pH indicator. Most organisms cannot grow in such a salty environment, but organisms of the genus Staphylococcus are uninhibited, making the agar selective for Staphylococcus. Therefore, if an organism is able to grow well on the agar, it is likely that it is Staphylococcus. The agar is differential because of the mannitol and the indicator. Only Staphylococcus aureus is able to ferment mannitol, which produces an acid end product. This acid accumulation will lower the pH of the agar plate, causing it to change colors to yellow. I was able to observe 3 organisms on a mannitol plate; one S. aureus that

Bailey, W. R., and E. G. Scott. 1966. Diagnostic microbiology: a text book for the isolation and identification of pathogenic microorganisms, 2nd ed. The C. V. Mosby Company, St. Louis, MO.

S. Aureus was the only bacteria species to turn coagulase-positive during the Tellurite-Glycine Test compared to all other tested species which had white colony coloration. No growth was reported after the first 24 hour check. After two days the colonies had all grown and S. Aureus was the only species with a faint coloring of black.

Professor handed out one unknown organism to each of the students. Students had to record their own unknown ID. Used the aseptic techniques to perform the gram stain and inoculate the nutrient media(nutrient broth, nutrient agar slant, and nutrient agar plate). Only for the nutrient agar plate media used four quadrant streak method. Gram stain was distinguishing the gram-positive or gram-negative cells. Preparing of the heat-fixed slide was taking some times while air dries it. Therefore, inoculated the media of growth during the waiting time. Additionally, placed the media in the “to be incubated area.” The gram stain required four important stains and solution: Crystal violet, iodine, 95% ethanol, and safranin. Crystal violet was for the primary stain. Iodine was the mordant to form the crystal violet-iodine complex. Used the 95% ethanol to decolorize the stained slide. Safranin was the counterstain to colorize the gram-negative cell. One had to use the DI water to rinse every stain between these procedures. It was very hard to decolorize the stains, because it may

When cultured on a Mannitol Salt Agar plate, we looked to observe if the isolate fermented mannitol. It was determined that the isolate ferments mannitol because of the yellow colonies that grew. The next test in this battery tested the presence of Catalase which is an enzyme that protects cells from damage by hydrogen peroxide by breaking it down into water and oxygen. The oxidase test which determines the presences of cytochrome oxidase determines if a bacterium can use oxygen as an energy source. The isolate tested positive for catalase but negative for oxidase. The final test to be conducted was to observe the possible lysis and degree of lysis caused by the unknown isolate. On a BPA plate, the unknown isolate lysed the surrounding red blood cells

From the oxidase test, unknown 65 appeared to be of the family Enterobacteriaceae. The Microbiology Laboratory Manual, chart II, showed very close matches for unknown 65 with Escherichia coli, as did Bergey’s Manual. Escherichia had identical tests results as unknown 65, except for urease and phenylalanine tests (appendix A-16). Also, E. coli is resistant to tetracycline and unknown 65 was not on the antibiogram (4). Unknown also matched Morganella morganii in all performed tests, except for the arabinose and lactose fermentation tests. Bergey’s manual showed the same results. The urease test was positive, and since this test can distinguish urease-positive Proteus, Morganella, Klebsiela, and Providencia from other Enterobacteriaceae, such as E. coli, which is urease negative, unknown 65 is likely to be Morganella (2). The phenylalanine test is also used to distinguish between the phenylalanine deaminase-positive genera Proteus, Morganella, and Providencia from other Enterobacteriaceae, such as E. coli, which is phenylalanine deaminase-negative (2). Considering these results and the lactose tests, which were contradictory, unknown 65 is most likely in the family Enterobacteriaceae, the genus Morganella, and the species morganii

To find the first unknown bacteria, the microbiology student started by inoculating a sample on a slide to stain with Gram’s stain; resulting a gram positive cocci. He continued by looking at the dichotomous key in the Lab Manual, afterwards doing a catalase test on an inoculated slide to differentiate it from being either a Streptococcus or a Staphylococcus (2). He then put the hydrogen peroxide on the inoculated slide, it immediately reacts, resulting on an excessive bubbling reaction, concluding that the Gram positive species is catalase positive. He now had to inoculate a sample of the bacteria to a 10% salt agar media to determine whether it is a Staphylococcus or a Micrococcus. After inoculating and incubating at 37°C for two days, it resulted with substantial growth. This indicates that the unknown Gram positive bacteria’s genus is a Staphylococcus. Following up, the microbiology student needed to determine whether it of the species aureus, saprophyticus, or epidermidis based on the selected characteristics table for staphylococcus (2). He conducted a hemolysis test on a thawed plasma plate; resulting in a β-hemolysis, a complete lysis of red blood cells. This test

Mannitol Salt Agar (MSA) medium has many components to differentiate S. aureus from other bacteria. The nutrients allowed bacteria to grow. The sodium chloride (7.5%) was the selective agent as most bacteria will be killed when exposed to excessive salt through dehydration. The carbohydrate mannitol acts as a differential agent as it is the substrate for fermentation to indicate between species of Staphylococcus. The phenol red indicator shows a change in the pH level. If acid is produced as an end-product due to fermentation, then the pH level will change.