MICROBIOLOGY It is very broad, not only bacteria but also parasites, bacteria, fungi and virus Science that

deals with the study of microorganisms using microscope

Terms: 1. Pathogens- microorganisms that can cause disease a. pathogenic- microorganisms that can cause disease b. non pathogenic- microorganisms that cant cause disease 2. Culture- microbes that grow and multiply in or on culture medium 3. Colony- visible growth of microorganisms deposited on the surface of a solid medium 4. Culture media- materials that help bacteria in the specimen to grow i. Food and soil of the bacteria ii. Contains nutrients for the bacteria Example: urine -> urinary tract infection; causative agent: Escherichia coli Best culture media for growth is eosin methylene group List of Common Culture Media and uses: 1. Modified Thayer Martin- used for the isolation of Neisseria gonorrhoeae; causative agent of gonorrhea 2. MacConkey agar- used for the differential isolation of gram negative bacilli and differentiation of lactose fermenters from non lactose fermenters i. Color pink = lactose fermenters ii. Colorless= non lactose fermenters iii. Salmonella shigella agar- combination of lactose and non lactose 3. Eosin- Methylene Blue agar- same function as MacConkey but selective medium of E. Coli a. Uninoculated- unused culture media (no growth of microorganism yet) *greenish metallic sheen- E. Coli is present * Klebsiella- siponish/ Flemish characteristic (large mucus colonies) 4. Bismuth Sulfite Agar- used for the selective isolation of salmonella from stool; color yellow a. Growth of salmonella- it is a hydrogen sulphite producer therefore can see black colonies 5. Blood agar- used for cultivation of fastidious organisms and determination of haemolytic reactions a. Non fastidious- very easy to grow; 37oc in incubator for 18-24 hours tutubo agad b. Fastidious- need enriching substance to grow Example: blood, serum Base: nutrient agar/ tripticase soy agar; color: yellow a. Blood agar- allow to cool then add blood

6. 7. 8. 9. 10. 11.

12. 13. 14.

Chocolate agar- used for the cultivation of haemophilus and Neisseria a. when nutrient agar/ base is still hot, add the blood Cystine-Tellurite blood agar- used for the reaction of Corynebacterium diphtheriae Fletchers semisolid medium- for Leptospira Lowenstein- Jensen medium- used for the cultivation of Mycobacterium tuberculosis Cetrimide agar- selective isolation and identification of Pseudomonas aeruginosa Mannitol Salt agar (MSA)- used for the selective isolation of pathogenic staphylococsi a. Staphylococcus aureus- pathogenic; colonies are yellow halo meaning mannitol is fermented b. Staphylococcus epidermidis- non pathogenic; colonies are pink meaning mannitol salt is not fermented Salmonella Shigella Agar- selective medium for the isolation of salmonella and shigella; color is peach Xylose- Lysine Desoxycholate (XLD)- isolation of enteric pathogens specially shigella; color is red orange Thiosulfate Citrate Bile Salt Sucrose Agar (TCBS)- isolation of Vibrio; color is green a. Vibrio cholerae- causative agent of cholera

NOSOCOMIAL INFECTION- hospital acquired infection Example: O.R infected ng pseudomonas, must always close the OR Opportunistic bacteria- when given opportunity they will cause a disease (mabababa immune system) MODE OF TRANSMISSION 1. Person to person- direct contact with patients who are ill 2. Contaminated medical equipment- dirty equipment like instruments and needles 3. Failure to follow established procedures to avoid spreading microbes 4. Bacteria environment (airborne transmission of microorganisms) TB- pwede airborne transmission 5. Open wounds Strict with students in laboratory; must wear lab gown, mask, etc. Proper attire cause if may open wound pwede pumasok sa sugat 1:10 dilution of Lysol for 10 mins before remove agar

CLASSIFICATION OF CULTURE MEDIA 1. Physical state or consistency a. Liquid/ Broth- contains no agar; must not solidify 1. Example: nutrient broth b. Solid- contains 1.5 to 3% agar c. Semi solid- .5 to 1% agar

1. Ex. Butt tube medium- to determine motility of bacteria 2. Application/ function a. Simple/ basal/ ordinary- to support growth of non fastidious bacteria 1. Example: nutrient broth and nutrient agar b. Enriched- growth of fastidious bacteria 1. Example: chocolate agar and blood agar c. Differential- differentiate bacteria from one another 1. Contains an indicator to differentiate 2. Example: eosin methylene agar- indicator is eosin methylene blue, mannitol salt agar- indicator is phenol red, Simmon- citrate agar- indicator is bromthmol blue( to determine if bacteria used citrate as sole source of carbon d. Selective- allows the growth of bacteria but suppresses the growth of other bacteria 1. Contains an inhibitory agar 2. Example: salmonella shigella agar 3. Composition a. Synthetic- exact composition is known b. Non synthetic- exact composition is not known 4. Form or distribution a. Tubed- displaced sa tube 1. Butt 2. Slant 3. Butt- slant 4. Broth (liquid) b. Plated *** TURBIDITY indicates growth of bacteria SPECIMEN COLLECTION REQUIREMENTS FOR SPECIFIC SITES Respiratory Tract a. Specimen from upper respiratory tract i. Throat cultures ii. Nasopharyngeal cultures iii. Specimens for oral cavity 1. Throat Cultures Use sterile cotton swab which is properly attached to applicator stick Requested to diagnose group A Sterptococcal infection casue by Streptococcus pyogenes (scarlet fever) Avid touching uvula; use tounge depressor to see i. Sterile cottonswab ii. Tounge depressor iii. Inoculate culture media iv. incubate

May also be requested for the following infections: i. Whopping cough- Bordetella pertussis ii. Epiglottitis- Haemophilus influenzae iii. Oral gonorrhea- Neiserria gonorrhoeae

2. Nasopharyngeal cultures Preferred for the diagnosis of pertussis Used to diagnose middle ear infections Maybe used to identify the carrier state of Staphylococcus aureus or Neiserria meningitidis 3. Lower Respiratory tract cultures Used to diagnose bronchitis or pneumonia Sputum: used to diagnose infections of the lower respiratory tract; larger volume in the morning; collected by dip coughing?? / sticky compared to saliva cause saliva is bubbly Used to diagnose Mycobacterium tuberculosis 4. Gastrointestinal tract Used to diagnose the cause of gastroenteritis stomach 5. Urine Used to diagnose UTI of the lower and upper tract Lower involving the bladder (cystitis); urethra (urethritis) Upper urinary tract infections include infections of the kidney such as pyelonephritis and glomerulonephritis Renal pelvis can be involved (pyelitis) Early morning urine and mid stream is best; aliquot portion- swab with applicator stick 6. Blood Used to diagnose bacteremia (presence of bacteria in blood) Thioglycolate- transport media; incubate 18-20 hours then inoculate in chocolate or blood agar Cinical isolate: specimen obtained from the patient Gram negative bacteria- habitat is gastrointestinal tract Venus blood- any vein (venipuncture) Arterial blood- interns dont do it cause it s a different way of collecting; blood from artery is super red Femoral tap- kuha ng blood from the singit Jurugal vein- neck kuha ng blood 7. Cerebrospinal fluid Collected by physician Requested to diagnose Neisseria meningitidis and Haemophilus influenzae