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Microbiology case study #9

Transcript: The symptoms can indicate a parasitic infection: -diarrhea -headache -fever -malaise Recent camping trip Possibly ingesting raw or undercooked meat Question 7 SOCIAL HA Negron, Simona Social The medical technician processing the culture should recognize these potential pathogens by: Escherichia coli O157:H7: -SMAC: NSF, positive motility Salmonella: -MAC: NLF (clear),HEK:NLF (green) with or without H2S,XLD:NLF(Red) with or without H2S Shigella: - MAC: NLF(clear) HEK:NLF(green) ,XLD:NLF(red) Campylobacter: - CAMPY agar: wet grey colonies Bacillus cereus: - BAP: large, dry colonies Plesiomonas shigelloides: - MAC: NLF,HEK:NLF,oxidase +,indole+ Aeromonas species: - BAP: Beta-hemolytic colonies, gram stain and oxidase+, indole+ Yersinia enterocolitica: - MAC: NLF Vibrio species: - TCBS: yellow/ green colonies MLT Phase II Camplyobacter jejuni is the most common of Campylobacter species foodborne diarrheal disease in U.S. Transmission is by consumption of contaminated food or water or improper preparation of poultry. Trichinella What parasites might have the physician suspected? * Vibrio species PARASITES Based on the history and laboratory results presented ,what is the most probable cause for the diarrheal disease in these patients? The couple denied drinking any water from the lake near their campsite, although after further questioning they admitted to skinny-dipping in the lake one evening “before the weather got bad.” Diane mentioned that although this trip was Jack’s idea of roughing it” they were careful to use only bottled water for drinking. She went on to explain that they bathed and used the sanitary facilities provided at the campsite and used only potable water from that facility for washing dishes and cooking. They described the food they had eaten during their trip: including hamburgers, hotdogs, chicken, roasted corn, canned beans, macaroni salad, and coleslaw. They transported the meat frozen and were careful to keep all the food on ice as much as possible. Diane mentioned that she thought the chicken they ate for lunch their last day may have been undercooked, but she didn’t want to make Jack go back out into the rain to cook longer. Symptoms: -Fever (Jack 100.9°F, Diane -100.2°F). -Headache -Myalgia -Malaise -2 to 3 loose bowels movements -7 to 9 watery bowel movements -Severe cramping Question 6 What bacterial pathogens should be included in the screening of a routine stool culture ,and how would the medical laboratory technician processing the culture recognize these potential pathogens? Why might the physician suspect that parasites could be a possibility in these patients? Questions . Ancylostoma duodenale . Patient History * Yersinia enterocolitica Jack and Diane R., a 25-­year old newlywed couple, reported to the emergency room with abdominal pain and diarrhea of 4 days’ duration. They stated that they initially thought they “picked up summer stomach flu” because they recently returned from a 5-­ day camping trip. Jack added, “We were really roughing it Doc. It was great fun until the weather got miserable, rainy and cold the last two days of the trip!" Patient profile: Necator americanus Other more unusual bacterial pathogens that may also cause diarrheal disease are Plesiomonas species Aeromonas species Escherichia coli Yersinia species Vibrio species Clostridium species Based on the history and laboratory results presented the most probable cause for the diarrheal disease in these patients Campylobacter species Entamoeba histolytica San Diego, CA Enterobius vermicularis Question 3 Giardia lamblia NO? GOOD! Question 1 27Oct2017 The bacterial pathogens should be included in the screening of a routine stool culture are: Escherichia coli Salmonella Shigella Campylobacter Bacillus cereus Patient History The specimen should be accessioned and processed ASAP 4 Quadrant streak on each of the following plates: BAP(Blood Agar Plate) -35 degrees Celsius: Ambient air MAC( MacConkey Agar) -35 degrees Celsius: Ambient air S-MAC(MacConkey Agar w/ Sorbitol) -35 degrees Celsius: Ambient air HEK(Hektoen Enteric Agar) -35 degrees Celsius: Ambient air XLD(Xylose Lysine Deoxycholate Agar) -35 degrees Celsius: Ambient air CAMPY(Campylobacter) -42 degrees Celsius: Microaerophilic Question 5 When a stool specimen is bloody ,additional testing is often recommended or suggested to the physician (especially with children or the elderly).What pathogen is of concern in that situation ,and how is this specimen processed? * Plesiomonas shigelloides Question 4 Lab Results Bacteriology plates: References Answers Lab Results QUESTIONS? Physical Examination -slight dehydration -elevated temperature -diffuse abdominal tenderness upon

Microbiology Case Study

Transcript: "Positive" aerobic: GS BAP CHOC MAC "Positive" anaerobic: GS BAPx2 CHOC MAC Patient Profile Patient Physical Exam Otto, M. (2009). Staphylococcus epidermidis - the "accidental" pathogen. Retrieved November 28, 2016, from If nothing grows for 5 days, the sample is discarded Instructional Aids Three subsequent blood culture were all negative Fever dropped and the patient's mental status cleared Patient was discharged 7 days later Reporting requirements/ Instructional aids Catalase positive Coagulase negative (both slide/tube) Sensitive to novobiocin This test is very important to distinguish it from S. saprophyticus; which it is also coagulase negative, but novobiocin resistant. Vital signs: Temperature: 38.9°C (high) BP: 94/60 mmHg (low) Pulse: 95 b/m (normal/high) Respiration: 23 breaths/min (high) Day 1: Urine and respiratory cultures: negative Blood cultures: 1 set positive Gram-positive cocci in clusters (doctor was notified) Day 2: *doctor removed Picc line and sent to microbiology laboratory Day 3: Blood culture: 1 set positive with S. epidermidis Port sample: positive GPC * PT was started on vancomycin Microbiology Case Study Instrumental Aids Blood culture was collected from PT's Picc line Blood (organism in blood is always critical) Report gram stain to the doctor within 30 minutes of sign of growth in blood culture. Report to the doctor the organism found. Differential Diagnosis References HN Renata Choi Because mac agar only grows GN organisms Catheter Report to the doctor that there was growth on the culture A venous access port is removed, and the port is sent to microbiology laboratory. It forms white, raised, cohesive colonies about 1-2mm in diameter after overnight incubation. Cheung, G. Y. (2010, October 7). Staphylococcus epidermidis Strategies to Avoid Killing by Human Neutrophils. Retrieved November 28, 2016, id=10.1371/journal.ppat.1001133 GS: Gram positive cocci Blood agar: Small white non-hemolytic Catalase: positive Slide Coagulase: Negative Tube Coagulase: Negative Novobiocin: Susceptible Test Summary Reporting Requirements Physical Exam: Erythema, warm, tenderness R clavicle Paleness Fatigue More About S. epidermidis NO grown on the macconkey agar Patient's workup Signs/Symptoms: skin red, painful and swollen warm to the touch Laboratory Testing Class 115 A 65 year old male presented to the emergency department with fever, altered mental status, and joint pain. S. epidermidis is a part of human normal skin flora; it is often attached to the upper layer of the skin (epidermis), without causing any symptoms. When the skin is injured (burn, wounds), the organism may enter into deeper layers of the skin or even in blood causing infection. It is not haemolytic on blood agar. Blood culture Leukocytosis (White blood cell count of 20,410/ul) High neutrophil counts (10,880/ul) High level of C-reactive protein (CRP: 6.39 mg/dl) Staphylococcus epidermidis is Gram-positive and it's in cluster forming cells that are a circular shape (cocci). Diagnostic Testing/Test Results Staphylococcus aureus Past Medical History S. aureus has long been recognized as one of the most important bacteria that cause disease in humans. PT stated that he recently changed his home nurse 1995: Diagnosed with hypertension 2014: Colectomy Diagnosed with colon cancer Picc line placed for chemo therapy 2015: Chemo therapy O'Gara, J., Humphreys, H. "staphylococcus epidermis biofilms: importance and implications". Journal of Medical Microbiology. 2001. Volume 50. p. 582-587. Patient's Information Draw blood 8-10 ml blood/bottle aerobic & anaerobic Place bottles in incubator; automatically monitors for growth * A healthy human's blood should be sterile Habitat: skin and mucous membranes Cell wall: glycerol-teichoic acids Virulence factor: "slime" Mod of transmission: implantation of medical devices such as catheters, shunts, and prosthetic devices. S. epidermidis causes biofilms to grow on plastic devices placed within the body. Infections are acquired nosocomially Serious infections among immunosuppressed patients may occur

Microbiology Case Study

Transcript: Stains the peptidoglycan layer of gram-positive bacteria (purple) Thinner layer of gram-negative bacteria is stained (pink) Bigger molecule than crystal violet Insoluble Binds to crystal violet It dehydrates the peptidoglycan layer It shrinks and tightens it For Gram positive bacteria, the big complex isn't able to penetrate this tightened layer and is trapped in the cell For Gram negative bacteria, the outer membrane of the cells are degraded and the thinner peptidoglycan layer of Gram negative cells doesn't retain the complex and color is lost Counterstain Does not disrupt purple colorization of Gram-positive bacteria Stains Gram-negative bacteria red Selective media inhibit growth of select microorganisms Differential media distiguishes growth between microorganisms growing on the same media MacConkey Agar Grows only Gram negative bacteria Contains bile salts which inhibit most Gram positive bacteria (selective) Distinguishes between bacteria that can and cannot ferment lactose Bacteria that can appear red/pink Bacteria that cannot appear white/colorless or golden to brown with dark centers Antimicrobials inhibit or kill microorganisms Antibiotics are antimicrobials made by microorganisms Trying to find out whether our bacteria sample is sensitive or resistant to different antibiotics Using the Kirby-Bauer method A plate covered with bacteria is used Antibiotics on paper discs are placed on the plate Zone of inhibition is the space around each disc that the bacteria cannot grow around Measuring the diameter of the zone of inhibition can allow us to determine if a bacteria is resistant or sensitive to an antibiotic Rinsing Out with DI Water Compare measurements found with measurement standards Positioned within 5 feet of each other Had separate filtration systems Pool: Automated chlorination system Hot tub: Chlorine and pH maintained manually Chlorine levels were low during outbreak 3 Water Drops on Slide Gentamycin is the most effective for treating the patient. Rinse with DI Water E. coli Placing disc on plate By Emily and Preethi Rod-shaped Gram-negative since it is pink Add Gram's Iodine Gram-negative Bacillus (rod) shaped Single bacillus arrangement Found in soil, water, skin flora Thrives on most surfaces Opportunistic, nosocomial pathogen Measure the zones of inhibition Obtain Sample Thick layer of peptidoglycan Folliculitis We know that our bacteria is Gram-negative and rod-shaped. Teichoic acids Known Gram-negative Stained pink Rod-shaped Selective and Differential Media Completed plates prior to incubation Ethyl alcohol Possible Bacterial Culprits View Under Microscope Known gram-positive Stained purple Spherical Case Study 2: Don't Swim in the Pool Measurements Antimicrobial Susceptibility Test Safranin Various people stayed at Hotel A in Bangor, Maine Stayed Feb 18 - Feb 27, 2000 Spent time in pool and/or hot tub 9 case patients, 7 spent time in both Adding Safranin Completed plates after incubation Pseudomonas aeruginosa MacConkey Agar sample after incubation Gentamycin Porins Iodine Solution Prepare Slide Used to treat many bacterial infections Particularly effective against bacterial infections from Gram-negative bacterias Synthesized by Micromonospora, which is a Gram-positive bacteria Administered intravenously, intramuscularly, or topically for infection treatment Gram + Background Rinse Off with DI Water (cc) image by nuonsolarteam on Flickr Single peptidoglycan layer Heat Fix the Samples Mixing in Bacteria Aminoglycoside Gram Staining the Sample Skin rash (folliculitis) Outer ear infection (otitis externa) Spreading bacteria on plate with cotton swab Can disrupt the bacterial cell membrane's integrity Might inhibit ribosomal translocation (from A-site to P-site) Can increase errors in synthesis of proteins, resulting in early termination Crystal Violet Dye Pseudomonas aeruginosa is green due to the presence of pycocyanine, a water-soluble green pigment. Outer membrance rich in lipopolysaccharide Allow Slide to Dry Our sample bacteria is green. Therefore, our bacterial culprit is Pseudomonas aeruginosa. Side Effects Otitis Externa Periplasm Gentamycin is ototoxic and nephrotoxic Can damage the vestibular apparatus in the ear with prolonged use Can sometimes cause complete hearing loss when there is a regularly harmless mutation in the mitochondrial RNA Can inhibit the synthesis of proteins in the renal cells (acute renal failure) General symptoms: Difficulty balancing Tinnitus Unsteady vision Psychiatric symptoms Confusion Depression Visual hallucinations Add Crystal Violet Patient Symptoms Retrieving a paper disc Pool and Hot Tub Gram - S. aureus Sample No outer membrane

Case Study: Microbiology

Transcript: CASE STUDY: MICROBIOLOGY By: Anand Arikarevula and Maya Rodgers Introduction As of March 7th, 425 persons from 44 states were reportedly ill with symptoms that included diarrhea, fever, and abdominal cramps The illness usually lasted 4 to 7 days Among 351 patients, 71 (20%) were hospitalized No deaths have been attributed to this infection Onset datesranged from August 1, 2006 to February 16, 2007 67% of these illnesses began after December 1, 2006. On March 16th, a patient comes to a clinic complaining of stomach cramping, diarrhea, and a moderately high temperature Had consumed a sandwich made with the peanut butter In order to identify the etiological agent, a fecal sample was taken and cultured Possible Causes: Escherichia coli Pseudomonas aeruginosa Salmonella enterica Staphylococcus aureus Streptococcus pyogenes Haemophilus influenzae Gram Stain & Selective and Differential Media Gram positive and Gram negative bacteria are types of bacteria based on the outer membrane Gram positive bacteriahave a thick peptidoglycan cell wall that lacks a lipopolysacharide envelope When a gram stain is done this type of bacteria will show a purple stain Gram negative bacteria contains a thin layer of peptidoglycan enveloped by a lipopolysacharide membrane. In a gram stain this type of cell will lose the purple stain in the beginning and go pink after the counter dye is added Procedure: Obtain a clean slide and make three drops of water on the slide Aseptically, use a loop add the three bacterias to the slide Air dry the slide and after heat fix it Cover the smear with crystal violet. Let it stain for one minute and then rinse it off with water Cover the smear with the iodine for one minute. After remove the iodine using water Tip the slide to a 45 degree angle and add the alcohol. Decolorize for 15-20 seconds. Remove the alcohol with the water Cover the smear with safranin for one minute and rinse off with water Observe under microscope Selective media are media which are formed to allow some microorganisms to grow while inhibiting others through the use of chemicals In the right concentrations, antimicrobial chemicals, such as antibiotics, will select for the growth of some organisms over others Differential media are formed to allow for the differentiation of microorganisms based on differences in the physical appearance of the colonies, or changes in color or opacity to the growth medium Based on the results of the Gram Stain, we chose the MacConkey Agar. MacConkey Agar is designed to select for Gram negative bacteria while differentiating between bacteria that can ferment lactose and those that cannot The Bile salts and Crystal Violet inhibit Gram positive bacteria, selecting for Gram negative The lactose present on the plate can be degraded by some bacteria into acidic products. If the bacteria can process lactose, the pH indicator in the agar changes color to hot pink. Antimicrobial Susceptibility Test The purpose of this test is to check which antiobiotic is susceptible to the bacteria. The antiobiotics used are: Penicillin Vancomycin Tetracyline Gentamycin Streptomycin Chloromphenicol Procedure: Use a sterile swab and make a lawn of bacteria for E.coli and the Patient sample Use the tweezers and place the 6 antimicrobial discs on the agar plate Make sure the discs are in good contact with the plate Incubate Results: Patient S. aureus: RESULTS Possible Causes: Escherichia coli Pseudomonas aeruginosa Salmonella enterica Staphylococcus aureus Streptococcus pyogenes Haemophilus influenzae Since we know our bacteria is Gram negative, that leaves us with: Escherichia coli Pseudomonas aeruginosa Salmonella enterica Haemophilus influenzae After placing the bacteria on the MacConkey Agar the bacteria did not produce lactose. This leaves us down to: Salmonella enterica Pseudomonas aeruginosa Haemophilus influenzae Now looking back at the symptoms of the patient, he had: Stomach cramping Diarrhea Moderately high temperature Therefore it is concluded that Salmonella enterica is the etological agent. The patient should take chloromphemnicol or tetracylcine as they are the most effective in killing the bacteria. Chloramphenicol works by inhibiting the enzyme peptidyl transferase, which prevents the synthesis of proteins Haemophilus influenzae symptoms are: Salmonella enterica symptoms are: Causes respiratory tract infections Tetracycline works by preventing tRNA from binding during protein synthesis, which prevents the addition of amino acids to the peptide chain. Commonly spread in water, causes lung and kidney failures Usually spread through poultry products, can cause diarrhea and other problems of the digestive tract Pseudomonas aeruginosa symptoms are:

Microbiology Case Study

Transcript: 4. What other nonpathogenic organisms commonly found in this type of specimen also cause this type of hemolysis? Blood and urine cultures were obtained to tell if the organism has spread to other parts of the body. Since Patrick was admitted to the emergency room, blood and urine cultures are routinely collected. Alpha-Hemolysis 2. Based on the colony morphology and the Gram's stain, what organism is suspect as the cause of Patrick's pneumonia? THANK YOU! Vaccination. For example, the PCV13 or Prevnar 13. Common preventative measures shold take place, such as: wash your hands, eat right, get enough sleep, and stop smoking. No, this patient does not have polymicrobial pneumonia. Continued... 7. Should antimicrobial susceptibility testing be performed on this pathogen? If so, what antimicrobial agent(s) should be tested? If not, why not? -Yes. - A Kirby Bauer plate with an Oxacillin disk should be used to determine resistance to penicillin. - Another test used is the E-Test (Epsilometer test), to test susceptibility to penicillin as well as Ceftriaxone. 1. Based on the direct gram stain, what is the quality of this sputum specimen? Is this specimen of acceptable quality to provide clinically relevant information? 6. Organisms other than the predominant organism were seen in the Gram's stain and culture. Does this mean that the patient has polymicrobial pneumonia? Why are those other organsims present? -The quality of this sputum specimen is acceptable due to the amount of epithelial cells in the specimen. -For sputum to be an acceptable quality, no more than 25 squamous epithelial cells should be observed per low power field (lpf). Rare squamous epithelial cells: (<1/lpf) 8. What virulence factor does the pathogen possess that can help it evade the host's defense mechanisms? Patient Profile/History Microbiology Case Study #5 HN Carver MLT Class 035 The direct Gram Stain of the sputum specimen revealed the following: Many Neutrophils (>25/lpf) Rare Squamous Epithelial Cells: (<1/lpf) Many Gram-positive lancet-shaped diplococcic and cocci in short chains (>25/oil immersion field) Few Gram-negative diplococci (<10/oil immersion field. Few Gram-positive bacilli (<10/oil immersion) To differentiate between S. pneumoniae and S. viridans, an optochin test as well as a bile solubility test should be performed. After overnight incubation at 35 degrees Celsius in 5% to 7% CO2, a blood-agar plate inoculated with the specimen revealed a mixture of two colony types. Rare, non-hemolytic, tiny, white, dry-looking colonies were present. A predominance of small, wet-looking colonies were also seen, with a greening of the medium around them. -The urine culture showed no growth at 24 hours. All blood cultures were negative after 5 days of incubation. 9. What preventative measures can be used to prevent infection or reinfection of this pathogen? Clinical Laboratory Science Review, Patsy Jarreau, 2011 Streptococcus viridans This Streptococci can also consist of gamma hemolysis as well as alpha, therefore explaining the "Rare, non-hemolytic, tiny, white, dry looking colonies present." 3. What type of hemolysis is described by the term "greening" of the medium? References Streptococcus pneumoniae consists of a polysaccharide capsule that prevents phagocytosis by the host's immune cells. S. pneumoniae also consists of surface proteins that prevent the activation of complement. A 75 Year-old man, Patrick R., presented to the emergency room with fever, shortness of breath, chest pain, and severe, extremely productive cough. Patrick had been a heavy smoker for almost 50 years before he quit seven years ago, when he was diagnosed with emphysema. Patrick occasionally used oxygen because of his severe respiratory distress. A chest x-ray revealed a right lower lobe infiltrate, and Patrick was admitted to the hospital. Sputum, urine, and blood cultures were collected. Streptococcus pneumoniae Colony morphology: "Small, wet-looking, convex (crater form), entire-edged colonies were also seen, with a greening of the medium around them." Gram-stain: "Many gram-positive lancet-shaped diplococcic and cocci in short chains." -Other organisms present in this specimen can be normal flora that is common in

Case Study- Microbiology

Transcript: Bacillus (rod-shaped) The study determined the MIC and in vitro bacterial killing curves. Among the test strains, Lc. lactis SL3 showed the strongest anti- C. difficile activity. as a Pathogen Microbiology Case Study In Vitro Evaluation of Antimicrobial Activity of Lactic Acid Bacteria against Clostridium difficile (CDI) Standard treatment: Antibiotics -vancomycin; metronidazole -Inhibits nucleic acid synthesis Hey Jim had those! Clostridium difficile Enters body via ingestion of spores (through the fecal oral route) Meet Clostridium difficile image2316557-Hour-monster.aspx Watery Diarrhea Fever Chills Abnormal weight loss Jim's stool test showed presence of bacteria The infection was hospital acquired Unfortunately, this isn't Jim's first illness... Maybe there's a connection? Purpose of study: CDI has become a major problem in public health. Broad-spectrum antibiotics are limited. Probiotics are a useful, promising option. Four strains of lactic acid bacteria (LAB) were evaluated for their anti-C. difficile activity. Results support that probiotics are effective against C. difficile-associated gut disorders. Jim Co-cultures showed the abilities of various probiotic bacteria to decrease the number and virulence of C. difficile . Mechanism: LAB strains produce organic acid. For example, lactic acid triggers lower internal pH of the bacterial cell, which has a bacteriostatic or bactericidal effect. Gram positive Treatments of Disease Higher percentage of people carry the bacteria in the hospital Usually develops within a few months after a course of antibiotics. lessonchallenge09/lesson/ Pharmacology/Introduction_08_print Lee J., Chung M. and Seo J. In Vitro Evaluation of Antimicrobial Activity of Lactic Acid Bacteria against Clostridium difficile. 2013. Toxicol. Res. Vol. 29, No. 2, pp. 99-106 Symptoms include weight loss, diarrhea, and fever. Jim was hospitalized 6 months ago for a urinary tract infection (UTI) He was given Ciprofloxacin antibiotic after his UTI Results Raizel Lidsky and Jackie Benayoun Meet Jim Jim's not feeling too great Clostridium difficile Jim's Doctor Confirms... Opportunistic Pathogen infection Poor Jim... In the case of severe pain or organ failure: Surgery to remove diseased portion of colon Produces toxins and forms spores which contributes to its virulence Clostridium difficile Techniques: Viewed under the fluorescent microscope and analyzed using Flow cytometric measurements (FCMs). His signs and symptoms: Can be found in body's GI tract Obligate anaerobe 10x higher risk to develop in people age 65 and over (Jim is 65!)

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