1. TESTS/ DIFFERENTIAL
1.1. AGAR PLATES
1.2. BIOCHEMICAL
1.2.1. ACETAMIDE UTILIZATION
1.2.2. ACETATE UTILIZATION
1.2.3. BACITRACIN TEST
1.2.4. BILE ESCULIN AGAR
1.2.5. BILE SOLUBILITY TEST
1.2.6. BUTYRATE DISK
1.2.7. CAMP TEST
1.2.8. CETRIMIDE
1.2.9. CITRATE UTILIZATION
1.2.10. DECARBOXYLASE T ESTS (MOELLER ’ S M ETHOD )
1.2.11. DNA HYDROLYSIS
1.2.12. ESCULIN HYDROLYSIS
1.2.13. FERMENTATION MEDIA
1.2.14. FLAGELLA STAIN (WET-MOUNT TECHNIQUE)
1.2.15. GELATIN HYDROLYSIS
1.2.16. GROW THAT 42°C
1.2.17. HIPPURATE TEST
1.2.18. INDOLE PRODUCTION
1.2.19. C ATALASE TEST
1.2.20. LAP TEST
1.2.21. LITMUS MILK
1.2.22. LYSIN IRON AGAR
1.2.23. METHYL RED /VOGES -PROSKAUER (MRVP) TESTS
1.2.24. MICRODASE TEST
1.2.25. MOTILITY TESTING
1.2.26. MRS BROTH
1.2.27. MUG TEST
1.2.28. NITRATE REDUCTION
1.2.29. NITRITE REDUCTION
1.2.30. ONPG (O -NITROPHENYL -β-D-GALACTO PYRANOSIDE) TEST
1.2.31. OPTOCHIN TEST
1.2.32. OXIDASE TEST (KOVAC’S METHOD)
1.2.33. OXIDATION /FERMENTATION (OF) MEDIUM (CDC METHOD)
1.2.34. PHENYLALANINE DEAMINASE
1.2.34.1. This test is used to determine the ability of an organism to oxidatively deaminate phenylalanine to phenyl-pyruvic acid. The phenylpyruvic acid is detected by adding a few drops of 10% ferric chloride; a green-colored complex is formed between these two compounds.
1.2.35. PYR TEST
1.2.35.1. The PYR test is predominately used in identification schemes for gram-positive cocci. Presence of the enzyme L - pyrroglutamylaminopeptidase that hydrolyzes the L -pyrrolidonyl-β-naphthylamide (PYR) substrate to produce a β-naphthylamine. The β- naphthylamine is detected in the presence of N, N-methylaminocinnamal- dehyde reagent by the production of a bright red–colored product
1.2.36. PYRUVATE BROTH
1.2.36.1. This test is used to determine the ability of an organism to utilize pyruvate. This aids in the differentiation between Enterococcus faecalis (positive) and Enterococcus faecium (negative).
1.2.37. SALT TOLERANCE TEST
1.2.37.1. This test is used to determine the ability of an organism to grow in high concentrations of salt. It is used to differentiate enterococci (positive) from nonenterococci (negative). A heart infusion broth containing 6.5% NaCl is used as the test medium. This broth also contains a small amount of glucose and bromcresol purple as the indicator for acid production.
1.2.38. SPOT INDOLE TEST
1.2.38.1. This test is used to determine the presence of the enzyme tryptophanase. Tryptophanase breaks down tryptophan to release indole, which is detected by its ability to combine with certain aldehydes to form a colored compound. For indole-positive bacteria, the blue-green compound formed by the reaction of indole with cinnamaldehyde is easily visualized. The absence of enzyme results in no color production (indole negative).
1.2.39. TRIPLE SUGAR IRON AGAR (TSI)
1.2.39.1. Triple sugar iron agar (TSI) is used to determine whether a gram-negative rod utilizes glucose and lactose or sucrose fermentatively and forms hydrogen sulfide (H2S). TSI contains 10 parts lactose : 10 parts sucrose : 1 part glucose and peptone. Phenol red and ferrous sulfate serve as indicators of acidification and H2S formation, respectively.
1.2.40. UREA HYDROLYSIS (CHRISTENSEN’S METHOD)
1.2.40.1. This test is used to determine the ability of an organism to produce the enzyme urease, which hydrolyzes urea. Hydro- lysis of urea produces ammonia and CO 2 . The formation of ammonia alkalinizes the medium, and the pH shift is detected by the color change of phenol red from light orange at pH 6.8 to magenta at pH 8.1.
1.2.41. X AND V FACTOR TEST
1.2.41.1. Members of the genus Haemophilus require accessory growth factors in vitro. Some Haemophilus spp. require X factor (hemin) alone, V factor (NAD, nicotinamide-adenine denucleotide) alone, or a combination of both.
2. Rhodococcus spp.
3. CLASSIFICATION
3.1. GRAM POSITIVE COCCI
3.1.1. CATALASE POSITIVE
3.1.1.1. MODIFIED OXIDASE POSITIVE
3.1.1.1.1. Micrococcus
3.1.1.2. MODIFIED OXIDASE NEGATIVE
3.1.1.2.1. COAGULASE POSITIVE
3.1.1.2.2. COAGULASE NEGATIVE
3.1.2. CATALASE NEGATIVE
3.1.2.1. BETA HEMOLYTIC
3.1.2.1.1. Streptococcus pyogenes (Group A Strep)
3.1.2.1.2. Streptococcus agalactiae (Group B Strep)
3.1.2.2. ALPHA HEMOLYTIC
3.1.2.3. GAMMA/ NO HEMOLYSIS
3.2. GRAM NEGATIVE COCCI
3.2.1. DIPLOCOCCI
3.2.1.1. Neisseria spp.
3.2.1.2. Moraxella catarrhalis
3.2.1.3. Veilonella
3.2.2. COCCOBACILLI
3.3. GRAM POSITIVE BACILLI
3.3.1. Branching
3.3.1.1. Nocardia spp.
3.3.1.2. Streptomyces spp
3.4. GRAM NEGATIVE BACILLI
3.4.1. GROWS ON McConkey agar
3.4.1.1. Oxidase-negative:
3.4.1.1.1. Enterobacteriaceae
3.4.1.1.2. Acinetobacter spp.
3.4.1.1.3. Stenotrophomonas
3.4.1.2. Oxidase-positive
3.4.1.2.1. Burkholderia spp.
3.4.1.2.2. Achromobacter spp.
3.4.1.2.3. Alcaligenes spp.
3.4.1.2.4. Bordetella (non-pertussis)
3.4.1.2.5. Comamonas spp.
3.4.1.2.6. Vibrio spp.
3.4.1.2.7. Aeromonas spp.
3.4.1.2.8. Plesiomonas shigelloides
3.4.1.2.9. Chromobacterium violaceum
3.4.2. NO GROWTH on McConkey agar, Oxidase-positive
3.4.2.1. Sphingomonas paucimobilis
3.4.2.2. Eikenella corrodens
3.4.2.3. Weeksella virosa
3.4.2.4. Pasteurella spp.
3.4.2.5. Suttonella indologenes
3.4.2.6. Actinobacillus spp.
3.4.2.7. Kingella spp.
3.4.2.8. Cardiobacteriun spp.
3.4.2.9. Capnoocytophaga spp.
3.5. ANAEROBES
3.5.1. GRAM POSITIVE
3.5.1.1. Clostridium
3.5.1.2. Lactobacillus spp.
3.5.1.3. Actinomyces spp.
3.5.1.4. Arcanobacterium spp.
3.5.1.5. Propionibacterium
3.5.1.6. Bifidobacterium sp.
3.5.2. GRAM NEGATIVE
3.5.2.1. Fusobacterium
3.5.2.2. Bacteroides
3.5.2.3. Prevotella
3.5.2.4. Porphyromonas
3.6. With Special requirements for growth
3.6.1. Bartonella spp.
3.6.2. Afipia spp.
3.6.3. Campylobacter spp.
3.6.4. Arcobacter spp.
3.6.5. Helicobacter spp.
3.6.6. Legionella spp.
3.6.7. Brucelia spp.
3.6.8. Bordetella pertussis
3.6.9. Bordetella parapertussis
3.6.10. Franciscella tularensis
3.6.11. Streptobacillus moniliformis
3.6.12. Spirillium minus