Violet Stain Lab Report

Next I performed a KOH test to further confirm that my organism was a Gram-negative species. For the KOH test, I added 3 drops of 10% potassium hydroxide (KOH) to a small drop of distilled water onto a clean microscope slide, transferred a visible clump of organism to the KOH solution using my inoculating loop. I than mixed the cells into the solution using small, circular motions for 60 seconds and then lifted up the loop to look for what appears to be a “stringing” affect which means it’s confirmed that it is gram- negative species. Next, I created a streak plate using nutrient agar so that I could see pure culture of my organism. I aseptically obtained a loop full of my organism and gently inoculated one quarter of the nutrient agar plate by running the loop back and forth across the surface. I then flame sterilized the inoculating loop, allowing it to cool for 10 seconds, and then streaked the organism from quadrant I into quadrant II using a zigzag motion technique. I repeated those steps streaking from quadrant II to quadrant III and then streaking from quadrant III to quadrant IV. Once completed, I put the streak plate in the incubator at 37° for 24-48 hours. 48 hours later, I check my streak plate and it had a lot of growth on it. I was able to determine that the organism was definitely an off white color, opaque. The IV quadrant was the quadrant that best

The Gram stain was used to determine if the bacteria was gram positive or negative. A negative test shows a pink color and a positive test is a purple color. When a bacterium is negative it is because it has an outer membrane and a thin layer of peptidoglycan that is much harder to stain. A positive bacterium has a thick layer of peptidoglycan and no outer membrane that can be penetrated by crystal violet.

8. Discuss a possible mechanism of Gram Staining in terms of differences in structure and chemistry between the walls of gram-positive and gram-negative

The predictions that were made prior to the experiment being ran about the concentration of pigments in Drosophila were similar to the observed concentration of pigments. Once four fly heads of each pigment were crushed onto filter paper, chromatography was ran. After six hours of the chromatography being run, they were removed from the solvent and dried. Once the filter paper was dry, it was placed under a UV light for a better observation of the pigments present in the eyes of the fly. What was found in the white was much less than in the wild type due to the fact that in the white eyed fly, both enzymes that start the pigment production (the pteridine pathway and the ommochrome pathway) are blocked or changed so no pigments can be produced.

The interventions should be focused on supporting the failing system and include the following: “(1) fluid replacement, (2) airway management, (3) antibiotic therapy, and (4) use of vasopressor” (Latto, 2008, p. 197). Fluid replacement is necessary to expand the blood and plasma volume in order to provide the adequate tissue perfusion and oxygen delivery to the organs. Vasopressors (dopamine, norepinephrine, epinephrine vasopressin) should be used in case the fluid replacement therapy fails to maintain adequate arterial pressure (Latto, 2008). The target central venous pressure should be more than 8 mmHG, the target central venous oxygen saturation should be over 70%. Wide-spectrum antibiotic should be administered as soon as the blood cultures are taken in order to treat the cause of the disease. Serum lactate level should be measured and treated with fluids “if greater than 1.5 times the upper limit of normal” (Latto, 2008, p. 198). Lactate is a byproduct of anaerobic cell metabolism and is one of the indicators of inadequate tissue oxygenation related to sepsis. Moreover, the glucose level, hematocrit and hemoglobin should be closely monitored.

3- Both hydrocortisone (1st line) methylprednisolone (3rd line in patient with hypokalemia, and fluid overload) can be used for treatment of adrenal crisis, however, the patient was already on a high dose methylprednisolone and adding more to achieve mineralocorticoids activity might not be

Hemodynamic Monitoring and Fluid management is considerable aspect in managing patient with ARDS. Swan-Ganz catheter did not get better results than standard monitoring and is associated with increased incidence of complications. Therefore, the routine use of pulmonary artery catheter is not necessary. A fluid balance neutral or mildly negative is associated with better outcomes from shorter duration of mechanical ventilation and ICU days, but no evidence supports reduction in mortality rates. Restrictive fluid management is adopted widely among literatures.

Tiffany O’Connor 9/2017 Gram-positive and Gram-negative stain Purpose: Staining is done to differentiate between two varieties of bacteria, gram-positive and gram negative. Theory and Background: Gram-positive and Gram-negative represents whether a cell wall retains the primary dye color in the staining process after decolorization. The difference in the structure of the cell wall determine if this happens.

Sepsis should be to be considered a medical emergency and treatment should begin the minute a patient presents with symptoms. Treatment options may begin with applying oxygen to the patient. Providing oxygen to the patient can ensure the body has enough oxygen in its system to perfuse vital organs. Treating patients with intravenous fluids can not only hydrate the patient but help to bring up a patient’s blood pressure than make be low due to the sepsis. Intravenous fluids such as normal saline (a crystalloid fluids) are usually the fluid of choice when treating sepsis. Another group of fluids that may be administered are colloid solutions. Colloids solutions include albumin and dextran. Albumin can replace lost fluid and help restore blood volume. Is a plasma volume expander that can assist in treating hypovolemia? When patients have an unstable or low blood pressure it can lead to shock. “A 2006 study showed that the risk of death from sepsis increases by 7.6% with every hour that passes before treatment begins’ (Kumar, 2006, p.251).

Humidified oxygen may help facilitate easier expectoration versus standard oxygen. Chest physiotherapy should be taught and is often beneficial in helping patients breathe properly, control their breathing and optimize their posture to promote suitable lung expansion and expectoration. Nurses should assist patients cough and deep breathe at least every 2 hours. Alert patients can also use an incentive spirometer to assist with deep breathing and stimulate coughing. Encouraging patients to drink at least 2 liters of fluid daily to prevent dehydration unless contraindicated is important in helping thin secretions move from the airway. Intake and output, oral mucous membranes, and skin turgor should be monitored for adequate hydration, especially when fever and tachypnea are

At the same time over hydration is also potentially harmful in this patient population as it has been associated with increased likelihood of the development of ACS. Overaggressive fluid therapy also has been linked to almost two times the chances of developing organ failure and potential fatality. This explains the importance of an appropriate goal directed fluid management and patients who have been adequately fluid resuscitated have improved end organ perfusion and functional recovery. Fluid therapy should therefore be carefully tailored to the individual patient population. Hypertonic saline and colloid has been used with variable success to decrease the development of ACS in burn patients since it decreases the requirement of total intravenous fluid, and hence the use of hypertonic saline or albumin should be considered in patients with elevated IAP. Extravasation of fluid; however, is bound to occur to a variable extent due to massive circulating inflammatory mediators in the body and increased vascular permeability. Role of diuretics may be questionable as these patients are intravascularly dry. If the patient is hemodynamically stable a combination of colloid and diuretic may be considered to optimize the patients fluid administration. Also the use

Women also have less body water than men and increase body fat can decrease water in the body. Assess patient vital before, during and after administration of a crystalloid IV therapy. Continuously monitor labs values (i.e. serum electrolytes) and notify the PCP of critical lab value. The nurse should assess catheter site for signs of infiltration or other complication. Priority nursing diagnoses include fluid volume excess, deficient Fluid Volume or water intoxication. When caring for patients, the nurse should plan to monitor weight daily and monitor for edema and dehydration. Educate the patients and their families about signs and symptoms of volume overload and dehydration, and instruct the patient to notify the nurse if they develop breathing or if they feel their heart is beating very fast, difficulties, swallowing, feel dizzy or malaise. The nurse should also elevate for the effectiveness of

If there is question of the two types of bacterium being located in the specimen then the addition of the counterstain safranin. As the procedure continues alcohol is introduce to the sample to differentiate the Gram-negative and the Gram-positive bacterium with it causing the outer layer of the Gram-negative cells to dissolve allowing the counterstain to remain to mask to crystal violet color making the cells show as pink or red vs. purple. In particular cases of staining of Bacillus, some Gram-positive cells will give a Gram-negative response though they are dead, there can be increasing numbers of Gram –negative cells; this is called Gram-variable. (Tortora, 2014) The procedure relating to staining for observation of Mycoplasmas involve a detailed and specific protocol for success. As previously mentioned, Mycoplasmas have little to no cell wall; the staining process has to be relative to what can be stained for observation. In my research, I found that the use of a specific stain, the Hoechst #33258 dye, is used to illuminate by florescence the DNA of Mycoplasma bacteria so it can be differentiated from the remainder of the sample by the use of a UV filtrated, florescent microscope. (Sciences, 2002) The opportunity of each of the given genera that could cause this particular

* Gram stain will be used to identify the gram bacterial membrane reaction for easy identification.

– Monitor of serum electrolytes once or twice daily and supplement as needed to avoid hyponatremia