Irina Pogorelskaya 04/11/12 Section 05 Unknown #5

Abstract: Goal of this lab is to use qualitative tests to find taxonomic identification of unknown. TA made a broth culture for everyone, culture 5 was randomly selected by me should contain two unknowns that have to be identify. It contains one gram positive and one gram negative. By having three lab periods and using TSA, blood agar, SS, TSI and citrate agar two unknown was identified as Escherichia coli, Bacillus subtillis.

Introduction: This lab is supposed to test our knowledge that was gain trough out semester and two unknowns should be recognized trough series of tests. Each person choose the unknown solution, each solution contains gram positive and one gram negative. Trough out three weeks with a use of at least three different tests for each bacterium and without help of TA, out of 9 different bacterias to choose from, two of unknowns should be identified. Escherichia coli would be first bacteria to talk about. Found and isolated by Theodor Escherich, the genus Escherichia was named after him. It is gram-negative bacilli that come in pairs or single, it is anaerobic with metabolism that is both fermentative and respiratory. E coli lives in in the large intestine, but at the same time one of the most frequent causes of many common bacterial infections, pneumonia, including neonatal meningitis, cholangitis, urinary tract infection (UTI), bacteremia, and traveler's diarrhea, and many others.(5) Klebsiella pneumoniae is family Enterobacteriaceae and genus Klebsiella. Edwin Klebs German microbiologist found it in 19th century. They are gram-negative bacteria, rod-shaped, with a polysaccharide capsule, which provides resistance against host defense mechanisms. Common infections in humans include UTI, rhinoscleroma, community-acquired pneumonia, nosocomial infection, colonization and ozena.(10) Salmonella enterica rod-shaped, Gram-negative, has a flagella, facultative anaerobic, and causes diseases of the intestine. It was named after scientist who discovered it, Dr. Daniel Salmon. It causes salmonellosis: enteric fever or typhoid by getting into the bloodstream, and acute gastroenteritis caused by a foodborne infection.(8)

Yersinia enterocolitica is a gram-negative bacillus that belongs to the family Enterobacteriaceae and found in1934 by Mclver and Picke .It is a main cause of acute diarrhea, pseudo appendicitis, mesenteric lymphadenitis, and terminal ileitis. (4) Streptococcus pneumoniae it is gram-positive, catalase negative, alpha-hemolytic, anaerobicand belongs to genus Streptococcus. It is most common cause of community-acquired pneumonia (CAP), bacteremia, bacterial meningitis, endocarditis, peritonitis, sinusitis, septic arthritis, and osteomyelitis. (6) Staphylococcus aureus is gram-positive cocci, grapelike clusters that colonize the skin,
and catalase-positive. It is seems to be resistant to high salt concentrations, temperatures as high

as 50C and to drying. The main cause of MRSA and found to effect nasal cavity, the pharynx, gut, and vagina.(9) Streptococcus pyogenes is gram-positive cocci, beta-hemolytic, non-spore forming. It colonize the upper respiratory tract and skin. The most common diseases are toxic shock syndrome (TSS), cellulitis, necrotizing fasciitis and bacteremia. (3) Staphylococcus epidermidis is gram positive, coagulase-negative staphylococci has been steadily increasingly in recent years. Infections are found in prosthetic heart valves, shunts, prosthetic joints, catheters, and large wounds and associated with necrotic skin and lung infections. Responsible for causing septicemia, endocarditis, pneumonia and osteomyelitis and many others. (2) Bacillus subtilis is Gram-positive rods, positive for catalase capable of growth in the presence of oxygen, and has an endospore that helps to sustain any weather conditions.

Commonly used in laboratories and used for enzyme reproduction. Causes food poisoning, nausea and diarrhea. (7) Materials and Methods: First week: Unknown sample# 5 was obtained from TA. TSA was streaked and blood agar to determine the colonies from the tube than they were incubated for next period. Second week: Blood plate and TSA was used for gram staining first, to determine negative and positive bacteria. Than positive bacteria was placed on mannitol salt agar, and catalase experiment was performed with help of hydrogen peroxide. For negative bacteria SS plate was used streaks of bacteria was placed, citrate agar were bacteria was placed on top of agar and TSI where the agar was stub with bacteria and placed on top. Casein plate was used for both.(1) Unknown #5 Gram stain Gram positive cocci

Gram positive rods

Gram negative rods

Blood agar

EMB

Casein

N/A

lactose
A) E.coli

Bacillus subtitilis

no lactose
C) Y.Enterolitica

B) K.pneumonie

D) S.enterica

Catalase test
S .pneumoniae

SS no H2S H2S
S.enterica

+/- +

MSA + / -
S.epidermidis

Citrate A)- B)+ C)+D)-

S.pyogenes
S.Aureus Results: Lab2: Blood agar was showing small colonies with a halo which showed beta Hemolysis. TSA showed two of bacteria white colonies. Lab3: When results came back TSI for sample B was yellow no gas formation or black formation, citrate came out green. SS had a small dot of black. MSA plate showed positive result for manntol formation. Casein was positive for A and negative for B. Discussion: Blood agar seemed like a beta hemolysis colonies had halo around them and it was positive for catalase bubbles occurred after addition of hydrogen peroxide. When gram stain was done the gram negative pink rods was showing for negative bacteria and purple rods and cluster of cocci was for positive believing contamination occurred which made positive bacteria identification wrong. On TSA p[late was showing white colonies of different sizes. On third lab period MSA

plate showed positive results for mannol formation it had yellow halo around it by using flow chart it was determined to be Staphylococcus aureus, which was wrong, casein plate showed positive result for Bacillus subtillis, but because of contaminated gram stain made a wrong choice. SS showed positive results for lactose formation, but it was too small to determine, so by looking at slant made all the difference. Green on citrate agar with yellow TSI agar with a no gas formation shows to be Escherichia coli, which was a right bacterium.(1)

Work Cited: 1) Carson., Valerie A, 1st ed Lab Manual 2010 2)Davis, Charles Patrick., Staphylococcus (Staph Infection), http://www.emedicinehealth.com/staphylococcus/article_em.htm, WebMD2012 3)Khan, Zartash Zafar., Streptococcus Group A Infections , http://emedicine.medscape.com/article/228936-overview, WebMD LLC 1994-2012. 4) Khan, Zartash Zafar Yersinia Enterocolitica, http://emedicine.medscape.com/article/232343-overview WebMD LLC 1994-2012 5)Madappa, Tarun., Escherichia Coli Infections, http://emedicine.medscape.com/article/217485-overview, WebMD LLC 1994-2012 6)Muench, F Dawn., Pneumococcal Infections , http://emedicine.medscape.com/article/225811-overview, WebMD LLC 1994-2012. 7)Slonczewski, Joan., Microbiology: An Evolving Science (Second Edition) Nov 10, 8)Todar, Kenneth., Salmonella and Salmonellosis http://textbookofbacteriology.net/salmonella.html 2008-2012 9)Tolan, Robert W Jr., Staphylococcus Aureus Infection http://emedicine.medscape.com/article/971358-overview, WebMD LLC 1994-2012 10)Umeh, Obiamiwe., Klebsiella Infections http://emedicine.medscape.com/article/219907-clinical, WebMD LLC 1994-2012