Antibiotics

  • Bacitracin
  • Chloramphenicol
  • Erythromycin
  • Kanamycin
  • Nalidixic Acid
  • Nitrofurantoin
  • Novobiocin
  • Optochin
  • Penicillin
  • Streptomycin
  • Sulfamethoxazole
  • Tetracycline

Biochemical tests

  • Acid from glucose
  • Adonitol fermentation test
  • Antibiotic susceptibility tests
  • Arabinose fermentation test
  • Arabitol fermentation test
  • Arginine dihydrolase test
  • Bacitracin susceptibility test
  • Catalase test
  • Cellobiose fermentation test
  • Citrate utilization test
  • Coagulase test
  • Deoxyribonuclease (DNase) test
  • Dulcitol fermentation test
  • Esculin hydrolysis test
  • Fructose fermentation test
  • Galactose fermentation test
  • Gas from glucose
  • Gelatin hydrolysis test
  • Glucose fermentation
  • Glycerol fermentation test
  • Growth in KCN (potassium cyanide)
  • Growth on 6.5% NaCl
  • Growth on 7.5% NaCl
  • Growth on bile salts test
  • Hand Hygiene and Infection Control
  • Hemolysis on blood agar
  • Hydrogen sulfide (H2S) production test
  • Indole production test
  • Inositol fermentation
  • Kirby-Bauer Antibiotic Susceptibility Test
  • Lactose fermentation test
  • Lipase test (using corn oil)
  • Lysine decarboxylase test
  • Malonate utilization test
  • Maltose fermentation test
  • Mannitol fermentation test
  • Mannose fermentation test
  • Melezitose fermentation test
  • Melibiose fermentation test
  • Methyl red test
  • Motility test
  • myo-Inositol fermentation test
  • Nitrate reductase test
  • Novobiocin susceptibility test
  • OF glucose test
  • Optochin susceptibility test
  • Ornithine decarboxylase test
  • Oxidase test
  • Phenylalanine deaminase test
  • Raffinose fermentation test
  • Rhamnose fermentation test
  • Ribose fermentation test
  • Salicin fermentation test
  • Sorbitol fermentation test
  • Starch hydrolysis test
  • Sucrose fermentation test
  • Trehalose fermentation test
  • Urea hydrolysis test (Christiansen’s)
  • Voges-Proskauer test
  • Xylitol fermentation test
  • Xylose fermentation test

Gram negative bacilli

  • Buttiauxella agrestis
  • Cedecea davisae
  • Cedecea lapagei
  • Cedecea neteri
  • Citrobacter amalonaticus
  • Citrobacter amalonaticus biogroup 1
  • Citrobacter freundii
  • Citrobacter koseri
  • Cronobacter sakazakii
  • Edwardsiella hoshinae
  • Edwardsiella ictaluri
  • Edwardsiella tarda (wild type)
  • Edwardsiella tarda biogroup 1
  • Enterobacter aerogenes
  • Enterobacter amnigenus biogroup 1
  • Enterobacter amnigenus biogroup 2
  • Enterobacter cancerogenus
  • Enterobacter cloacae
  • Enterobacter gergoviae
  • Escherichia coli
  • Escherichia fergusonii
  • Escherichia hermannii
  • Escherichia vulneris
  • Ewingella americana
  • Hafnia alvei
  • Klebsiella oxytoca
  • Klebsiella pneumoniae subspecies ozaenae
  • Klebsiella pneumoniae subspecies pneumoniae
  • Klebsiella pneumoniae subspecies rhinoscleromatis
  • Kluyvera ascorbata
  • Kluyvera intermedia
  • Leclercia adecarboxylata
  • Mollerella wisconsinsis
  • Morganella morganii subspecies morganii
  • Morganella morganii subspecies sibonii
  • Proteus mirabilis
  • Proteus myxofaciens
  • Proteus penneri
  • Proteus vulgaris
  • Providencia alcalifaciens
  • Providencia rettgeri
  • Providencia rustigianii
  • Providencia stuartii
  • Rahnella aquatilis
  • Raoultella ornithinolytica
  • Raoultella terrigena
  • Salmonella bongori
  • Salmonella enterica
  • Salmonella enterica subspecies arizonae
  • Salmonella enterica subspecies enterica
  • Salmonella enterica subspecies houtenae
  • Salmonella enterica subspecies paratyphi
  • Salmonella enterica subspecies salamae
  • Salmonella typhi
  • Serratia ficaria
  • Serratia fonticola
  • Serratia marcescens
  • Serratia odorifera biogroup 1
  • Serratia plymuthica
  • Serratia proteamaculans subspecies proteamaculans
  • Serratia rubidaea
  • Shigella dysenteriae
  • Shigella flexneri
  • Shigella sonnei
  • Shimwellia blattae
  • Tatumella ptyseos
  • Yersinia enterocolitica
  • Yersinia frederiksenii
  • Yersinia kristensenii
  • Yersinia pestis
  • Yersinia pseudotuberculosis

Gram positive cocci

  • Dermacoccus nishinomiyaensis
  • Enterococcus avium
  • Enterococcus faecalis
  • Enterococcus faecium
  • Enterococcus gallinarum
  • Kocuria kristinae
  • Kocuria rosea
  • Kocuria varians
  • Kytococcus sedentarius
  • Lactococcus lactis
  • Lactococcus raffinolactis
  • Macrococcus caseolyticus
  • Micrococcus luteus
  • Micrococcus lylae
  • Nesterenkonia halobia
  • Rothia mucilaginosa
  • Staphylococcus aureus
  • Staphylococcus auricularis
  • Staphylococcus capitis
  • Staphylococcus caprae
  • Staphylococcus carnosus
  • Staphylococcus chromogenes
  • Staphylococcus cohnii ssp. cohnii
  • Staphylococcus cohnii ssp. urealyticum
  • Staphylococcus epidermidis
  • Staphylococcus gallinarum
  • Staphylococcus haemolyticus
  • Staphylococcus hominis
  • Staphylococcus hyicus
  • Staphylococcus intermedius
  • Staphylococcus lentus
  • Staphylococcus saccharolyticus
  • Staphylococcus saprophyticus
  • Staphylococcus sciuri
  • Staphylococcus simulans
  • Staphylococcus warneri
  • Staphylococcus xylosus
  • Streptococcus acidominimus
  • Streptococcus agalactiae
  • Streptococcus bovis
  • Streptococcus criceti
  • Streptococcus equi
  • Streptococcus equinus
  • Streptococcus ferus
  • Streptococcus iniae
  • Streptococcus mutans
  • Streptococcus pneumoniae
  • Streptococcus pyogenes
  • Streptococcus ratti
  • Streptococcus salivarius
  • Streptococcus sanguinis
  • Streptococcus sobrinus
  • Streptococcus uberis

Media

  • 6.5% NaCl broth
  • Arginine dihydrolase broth
  • Bile esculin agar slant
  • Blood agar
  • Christiansen’s urea broth
  • DNase agar with methyl green plate
  • Lysine decarboxylase broth
  • Malonate broth
  • Mannitol salt agar
  • MRVP broth
  • Mueller Hinton Agar
  • Nitrate broth with Durham tube
  • Nutrient agar plate
  • Nutrient broth
  • Nutrient gelatin
  • OF glucose broth
  • Ornithine decarboxylase broth
  • Phenol red adonitol broth
  • Phenol red arabinose broth
  • Phenol red arabitol broth
  • Phenol red cellobiose broth
  • Phenol red dulcitol broth
  • Phenol red fructose (levulose) broth
  • Phenol red galactose broth
  • Phenol red glucose (dextrose) broth with Durham tube
  • Phenol red glycerol broth
  • Phenol red inositol broth
  • Phenol red lactose broth
  • Phenol red maltose broth
  • Phenol red mannitol broth
  • Phenol red mannose broth
  • Phenol red melezitose broth
  • Phenol red melibiose broth
  • Phenol red raffinose broth
  • Phenol red rhamnose broth
  • Phenol red ribose broth
  • Phenol red salicin broth
  • Phenol red sorbitol broth
  • Phenol red sucrose (saccharose) broth
  • Phenol red trehalose broth
  • Phenol red xylitol broth
  • Phenol red xylose broth
  • Phenylalanine agar slant
  • Potassium cyanide (KCN) broth
  • Rabbit plasma
  • Simmons’ citrate agar slant
  • Spirit blue agar plate
  • Starch agar
  • Triple sugar iron agar slant
  • Tryptone broth

Reagents, Chemicals, Stains

  • Ammonium Sulfate (saturated)
  • Barritt’s A reagent
  • Barritt’s B reagent
  • Crystal Violet
  • Ferric chloride (10%) reagent
  • Gram’s Decolorizer
  • Gram’s Iodine
  • HCl 0.1N reagent
  • Hydrogen peroxide
  • Iodine reagent
  • Kovac’s reagent
  • Methyl red reagent
  • Nitrate A reagent
  • Nitrate B reagent
  • Oxidase reagent
  • Rabbit Plasma
  • Safranine

Nitrate reductase test

About this test

What is the purpose of the test?  

This test determines whether the microbe produces the enzymes nitrate reductase and nitrite reductase . The two enzymes catalyze two reactions involved in converting starting compound nitrate into end product nitrogen gas.

             nitrate reductase                     nitrite reductase

NO3 ———————————> NO2 ———————————–> N2

 nitrate                                     nitrite                                nitrogen gas

Some bacteria produce both enzymes, some produce nitrate reductase only, and others produce neither.

How is the presence of these enzymes determined?  

If a bacterium producing nitrate reductase is grown in a medium containing nitrate , the enzyme converts the nitrate to nitrite . Nitrite reacts with certain chemicals to yield a red-colored product. If the bacterium also produces nitrite reductase , nitrogen gas will be liberated. Bubbles collecting in an inverted Durham tube indicate that nitrogen has been produced.

What medium is used?  

The medium used is nitrate broth with Durham tube, a nutrient broth with potassium nitrate added. The inverted Durham tube is placed in the medium as a trap for any nitrogen that might be generated.

How is the test performed?  

An inoculum from a pure culture is transferred aseptically to a sterile tube of nitrate broth containing an inverted Durham tube. The inoculated tube is incubated at 35-37 C for 24 hours and the results are determined. A positive test for both enzymes consists of a turbid (cloudy) broth with pronounced gas bubbles trapped in the Durham tube. If results like this are not observed, testing for the individual enzymes can be done through addition of reagents, with a positive test indicated by the broth turning red.

What reagents are added?  

Five drops of nitrate reagent A is added, followed by five drops of nitrate reagent B .

Performing this test in the VUMIE Online lab

Inoculation of Medium

1. Select the nitrate broth with Durham tube medium.  

2. Complete the process of a tube-to-tube aseptic transfer to inoculate the medium.

Incubation of the Inoculated Medium

3. Place the inoculated tube into the 35-37 C incubator.  

4. Press the New Day button to move forward 24 hours.

Determination of Test Results and  Addition of Reagents

5. Incubate this test for only 24 hours.  

6. Retrieve the incubated culture from the incubator.

7. Observe the culture. If the test was followed as described above, the culture will be cloudy (turbid). If there are bubbles in the Durham tube, this indicates presence of both nitrate reductase and nitrite reductase (nitrates have been converted to nitrites, and then to nitrogen gas – positive for both enzymes).

8. If there are no bubbles in the Durham tube, it will be necessary to add reagents to determine the results.   Select the dropper tool and the appropriate reagent needed from the chemical shelf. Remove the cap lid.  For this test, select Nitrate reagent A, add it, and then add Nitrate reagent B.   

9. If the tube turns red, it is an indication that nitrate reductase is present but nitrite reductase is absent (+ for the first enzyme but — for the second).   If the color doesn’t change, and there was no gas in the Durham tube, the bacterium does not have either enzyme.

10. Record test results.

Uninoculated Nitrate Broth

Positive for Nitrate (NO3) Reductase

Negative for Nitrate (NO3) Reductase

Positive for Nitrite (NO2) Reductase (bubbles of N2 gas in Durham tube)

Negative for Nitrite (NO2) Reductase (no bubbles) 


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Updated on November 1, 2022
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