micro lab final presentation

THE ISOLATION, CHARACTERIZATION, AND IDENTIFICATION OF A BACTERIUM FROM:STAPHYLOCOCCUS EPIDERMIDIS

Taken from: Stillwell bathroom paper towel dispenser

Hailey Medder12/08/2016

CULTURE CHARACTERISTICSUnit Being Observed

Size Shape Color

Colony 1.0 millimeter Cocci (clusters)

Opaque cream

Cell 0.5-.0.7 micrometers

Spherical ‘points’

Opaque cream

Photo by: Brittany H. from StudyBlue “Microbiology Final”

Culture Growth CharacteristicsTemperature (oC)

Salt (%NaCl)

pH

Oxygen

-Does not need oxygen due to being a facultative anaerobe

45

15

4018-40

Salt% 1 2 3 4 5 6 pH0% + + + - - - 35% + + + + + + 510% + + + + + + 715% - - - + + + 9

Salt and pH Treatment

0.0 10

2.0 4.0

6.0

8.00.0-0.5

57.0-7.45

7.0

9.0

Enteropluri

Glucose fermentation: YesGas production:

PartialLysine decarboxylation: NoOrnithine decarboxylation: NoSulfur reduction to H2S:No

Indole produced: NoAdonitol fermentation: NoLactose fermentation: NoArabinose fermentation: PartialSorbitol fermentation: NoAcetoin production (VP):NoDulcitol fermentation: No

Phenylalanine deamination: NoUrea hydrolysis: YesCitrate utilization: No

Motility- No

Oxidase- No

Catalase- Yes

Source: ResearchGate

Source: Microbeinfo.com

Blood agar

Maconkey agar

Although there were tons of colonies present, the S. epidermidis did not hemolyze the blood agar.

There was absolutely no growth on this Maconkey agar.

Positive gram stain

DNA SEQUENCES: DATA FOR 16S RDNA ANALYZED USING THE RIBOSOMAL DATABASE PROJECT

Database Name % Match EnvironmentType Cultures Only

Staphylococcus epidermidis

99.3 Clinical isolates in a Japanese zoo

Staphylococcus capitis 97.3 Japanese veterinary school Staphylococcus capitis 96.4 Skin samplesStaphylococcus caprae 96.3 Japanese rRNA samplesStaphylococcus saccharolyticus

95.8 Tissue samples

All RDP RecordsStaphylococcus epidermidis

100 GenBank in Northern Ireland

Staphylococcus epidermidis

100 Topoisomerase of isolates

Staphylococcus sp. 100 Wastewater in IndiaStaphylococcus sp. 100 Bluegill mucus in JapanStaphylococcus epidermidis

100 GenBank in Madrid Spain

DNA Sequences: Data for 16s rDNA Analyzed using the Ribosomal Database Project

Phylum: FirmicutesClass: BacilliOrder: BacillalesFamily: StaphylococcaceaGenus: Staphylococcus

COMPARATIVE CHARACTERISTICS TABLE

Test S. aureus S. caprae S. haemolyticus

S. epidermidis

Growth at 40ºC + + + +Growth at pH 4 - - - -Growth in 2% NaCl + + + +Hydrolysis of Tween 80

- - - -

Hemolyze blood cells

+ + - -

Acid from glucose + - + +Acid from raffinose - - - -Triclosan resistnce - - - -Capsules present + + + +Nitrate reduction - + + -

HAVE YOU IDENTIFIED A NEW SPECIES OF BACTERIA?

No

No I have not identified a new bacterial species. I know this because the data found on this species was published in multiple scholarly articles as well as in the Bergey’s Manual dating back to the early 1880’s.

ABSTRACT

Cells are Gram-positive, non-spore-forming, non-motile spheres approximately 0.5 μm in diameter. Colonies are creamy, circular, smooth, and 1.0 μm in diameter after 48 hour incubation at 37°C on R2A. The species could grow over10 μm from a range of salt concentrations from 0-0.5%, from pH 7.0-7.45, and at 37°C. Cultures are facultatively anaerobic, catalase-positive, and oxidase-negative. Colonies are non-hemolytic. Acid is produced from d-glucose, maltose, and partially from l-arabinose, but not from glycerol, glycogen, inulin, d-lactose, d-mannitol, raffinose, d-ribose, d-sorbitol, sucrose, trehalose or d-xylose. Hippurate, gelatin, and aesculin are not hydrolysed but starch and urea are. It does not utilize citrate. Nitrate is not reduced. Acetoin production is negative. Cells produce capsules but not flagella. No H2S is formed metabolically. DNAse and lipase activity are not present. Cells are susceptible to penicillin. The 16S rDNA sequence obtained from the culture revealed it to belong to the phylum Firmicutes, class Bacilli, order Bacillales, family Staphylococcaceae, and genus Staphylococcus. The closest relative of the culture is S. capitis, which has a 97.3% sequence homology to the isolate.

The strain was isolated from the paper towel dispenser handle in Cullowhee, North Carolina and is given the name Staphylococcus epidermidis.

REFERENCESATCC, https://www.atcc.org/Products/All/CRM-12228.aspx#culturemethod

S., F. A., Buchanan, R. E., Gibbons, N. E., & B. (1975). Bergey's Manual of Determinative Bacteriology. Taxon, 24(2/3), 377. doi:10.2307/1218353

Staphylococcus Genus Characteristics, http://biolabs.tmcc.edu/Micro%20Web/Staph.pdf

IJSEM database

M. B. (2004, September 27). Staphylococcus epidrmidis. Staphylococcus Epidrmidis. Retrieved December 6, 2016, from http://web.uconn.edu/mcbstaff/graf/Student presentations/S epidermidis/sepidermidis.html

https://www.ncbi.nlm.nih.gov/pubmed/9554167

Questions? Please be kind!