Differential Staining Worksheet (5] Apply what you have learned about Differential Stains by completing this graded worksheet. This should be submitted via Bb by the assigned due date (can be handwritten, scanned, or typed). Although more diagnostic than simple stains, Gram stains are often used to assign a bacterial isolate to the most general of categories (Ex: Gram-positive or Gram-negative). This lack of specificity for other cell structures is why morphology and Gram staining techniques cannot be used alone to ID/ classify bacteria. However, employing other special stains may aid in ID/ diagnoses when paired with basic staining techniques. Case Study: A New York Hospital admitted two patients exhibiting similar signs and symptoms: fever, chill, chest pain, coughing, fatigue, and shortness of breath. Patient #1: A 42-year-old female maternity nurse fell ill with what was first diagnosed as asthma. Her chest X-ray was interpreted as normal, and her symptoms were treated with steroids, antihistamines, and a cough suppressant. Despite this treatment her symptoms persisted, and after approximately 3-weeks, she returned to the hospital and was diagnosed with a suspected Streptococcus pneumoniae infection and prescribed IV ampicillin. Unfortunately, she returned with the same complaints 2-weeks later and further tests were ordered including a CT scan and bronchoscopy. Differential stains of her sputum sample are below: Gram Stain Acid-fast Stain Endospore Stain los Patient #2: A 44-year-old male drummer for an African dance troupe collapsed after experiencing mild shortness of breath, cough, and malaise over a three-day span. A chest X-ray revealed bilateral infiltrates and pleural effusions which were drained and treated with IV cephalosporin and steroids. Unfortunately, the patient’s respiratory status continued to worsen, and differential stains of blood cultures revealed the following: Gram Stain Acid-fast Stain Endospore Stain 1. Looking at the differential stains performed on each patient sample, is it possible that they both have the same infection? Why or why not? Be specific. 2. Using the dichotomous key below, circle the most plausible causative agent for each case. (You may have to do a little background research on each bacterium…) Gram + Bacilli Acid Fast + Acid Fast – Spore – Spore + Genus Mycobacterium Bacillus Clostridium Species → leprae tuberculosis anthracis subtilis difficile tetani 3. Support your diagnosis for each patient by: a. Explaining how they may have come into contact with the causative agent (reservoir/ route of transmission…). b. Explaining why the primary antibiotics prescribed for each patient did not control the causative agent (selective toxicity…). 4. From an epidemiological standpoint, should the hospital be worried about community spread where either causative agent is concerned? Why or why not?

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