LaMotte POL-H User Manual

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LaMotte BioPaddles™ TECH DOCUMENT

Call: 800-344-3100

Email: tech@lamotte.com

BioPaddles™ Colony Identification App

SAB/SAB

Code 5551

USE:

Selective isolation of fungi (yeasts and molds) (SAB)

Side 1 & 2: Sabouraud Dextrose Agar (SAB) (colorless / slightly hazy)

APPLICATION

Sabouraud Dextrose Agar (SAB) is a modification of dextrose agar described by Sabouraud.1 SAB is used for cultivating pathogenic and commensal fungi and yeasts. The high dextrose concentration and acidic pH of the formula permit selectivity of fungi.2 Sabouraud Dextrose Agar is used for determining the microbial content of cosmetics,3 in the mycological evaluation of food,4 and clinically to aid in the diagnosis of yeast and fungal infections.5

PADDLE AGARS

Sabouraud Dextrose Agar (SAB) – Enzymatic digest of casein and enzymatic digest of animal tissue provide the nitrogen and vitamin source required for organism growth in SAB. The high concentration of dextrose is included as an energy source. Agar and a proprietary polymer are the solidifying agents.

CULTURE CONTROLS

10-300 inoculum (CFU)

SAB Agar Aspergillus niger GROWTH Candida albicans GROWTH Escherichia coli INHIBITED Microsporum canis GROWTH Trichophyton mentagrophytes GROWTH

Sabouraud, R. 1892. Ann. Dermatol. Syphilol. 3:1061.

Jarett, L., and A. C. Sonnenwirth (eds.). 1980. Gradwohl’s and parasitic infections, 7th ed. American Public Health Association, Washington, D.C.

Curry, A. S., J. G. Graf, and G. N. McEwen, Jr. (eds.). 1993. CTFA Microbiology Guidelines. The Cosmetic, Toiletry, and Fragrance Association, Washington, D.C.

U.S. Food and Drug Administration. 1995. Bacteriological analytical manual, 8thed. AOAC International, Gaithersburg, MD.

Murray, P. R., E. J. Baron, M. A. Pfaller, F. C. Tenover, and R. H. Yolken (eds.). Manual of clinical microbiology, 6th ed. American Society for Microbiology,

Washington, D.C.

For in vitro diagnostic use only. This product should be used only by adequately trained personnel with knowledge of microbiological techniques in the laboratory.

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Page 2

LaMotte BioPaddles™ TECH DOCUMENT

Call: 800-344-3100

Email: tech@lamotte.com

STORAGE / EXPIRATION

Store tightly sealed BioPaddles™ in a cool, dry location. Shield from direct sunlight. Store BioPaddles™ at room temperature (65 - 77°F/18 - 25°C). Avoid sudden temperature changes. Temperature fluctuations may result in condensation settling at the bottom of the vial. This will not affect the culture properties, but could reduce the shelf-life or cause the agar to separate from the plastic paddle support. Do not refrigerate. Refrigeration may result in water condensation. Avoid freezing. If freezing occurs, thaw for 3-6 hours in the refrigerator (40°F; 4.4°C). Freezing can promote excess water loss and variation in media surface due to crystal formation.

Refer to Best Before End date (See: BBE stamped on vial). Discard if paddle agar appears oxidized and darker than the expected color or if contaminants appear. The expiration date is based on medium in an intact container that is stored as directed.

SAMPLING

Liquids: Twist to remove paddle from vial. Fill vial to 40 mL fill line with the liquid to be sampled. The 40 mL volume can be used to calculate Total Viable Count (TVC) and/or Total Colony Count (TCC). Replace paddle. Allow a contact time of 15 seconds. Remove the paddle. Empty the vial. Replace the paddle in the vial.

Surfaces: Twist to remove paddle from vial. Allow the paddle surface (10 cm2) to come into physical contact with the test surface. Recovery rate is about 50%. To ensure an accurate recovery, gently sweep (or touch) the paddle to cover a 20 cm2 area. Replace paddle in vial.

INCUBATION

Temperature Minimum Period Optimal Period

35°C (bacteria) 72 hours 5-7 days 20-25°C (fungi) 5 days 7 days

COLONY MEASURING

Each BioPaddles™ paddle has molded media attachment points that are 4mm in length (point-to-point). This feature provides a useful guidepost to estimating nearby colony size.

For in vitro diagnostic use only. This product should be used only by adequately trained personnel with knowledge of microbiological techniques in the laboratory.

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IDENTIFICATION

ORGANISM

PHYSIOLOGY

◆

Precision Test Strip

Available

GROWTH

COLONY

LaMotte BioPaddles™ TECH DOCUMENT

Call: 800-344-3100

Email: tech@lamotte.com

SAB

IMAGE

B. subtilis

E. coli

lternaria spp.

spergillus niger

• Lactose (-)

• Indole (-) ◆

• Oxidase (-) ◆

• Catalase (+) ◆

• Urease (+) ◆

• Gram (+) Rod

• Lactose (+)

• Indole (+) ◆

• Oxidase (-) ◆

• Catalase (+) ◆

• Urease (-) ◆

• Gram (-) Rod)

• Catalase (+) ◆

• Ascomycete

•

Catalase (+) ◆

•

Ascomycete

+++

PARTIAL COMPLETE INHIBITION

+++

• Translucent to dull, off-white; opaque

• Smooth to rough

• irregular / dendroid margins

to spreading

• 2-4mm

---

• Downy to woolly; flat

• Grayish, short, aerial hyphae

• later becomes greenish black

or olive brown with a light border

• 2-5++cm

• Granular

• White, w/ jet black fruiting

bodies w/ yellow/gray hyphae

• 2-5++cm

---

For in vitro diagnostic use only. This product should be used only by adequately trained personnel with knowledge of microbiological techniques in the laboratory.

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Page 4

ORGANISM

PHYSIOLOGY

◆

Precision Test Strip

Available

GROWTH

COLONY

LaMotte BioPaddles™ TECH DOCUMENT

Call: 800-344-3100

Email: tech@lamotte.com

SAB

IMAGE

spergillus flavus

spergillus

fumagatus

• Catalase (+) ◆

• Ascomycete

• Catalase (+) ◆

• Ascomycete

+++

• Granular to wooly

• Yellow, yellow-green or

yellow-brown pigment

• 2-5++cm

• Granular to cottony

• Blue-green, green-gray,

green-brown pigment

• 2-5++cm

spergillus terreus

• Catalase (+) ◆

• Ascomycete

+++

• Granular, radially rugose (wrinkled)

• Cinnamon buff, brown pigment

• 2-5++cm

IMAGE PENDING

For in vitro diagnostic use only. This product should be used only by adequately trained personnel with knowledge of microbiological techniques in the laboratory.

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Page 5

ORGANISM

PHYSIOLOGY

◆

Precision Test Strip

Available

GROWTH

COLONY

LaMotte BioPaddles™ TECH DOCUMENT

Call: 800-344-3100

Email: tech@lamotte.com

SAB

IMAGE

Botrytis spp.

Candida albicans

Chaetomium spp.

• Catalase (+) ◆

• Ascomycete

• Catalase (+) ◆

• Ascomycete

• Catalase (+) ◆

• Ascomycete

+++

• Wooly

• white, grey/brown pigment

• 2-5++cm

• White to cream

• Smooth /Spreading

• 2-6mm

• Wooly

• white, grey/olive pigment

• 2-5cm

Cladosporium spp.

• Catalase (+) ◆

• Ascomycete

• Granular to wooly (velvety)

• Olive-brown to blackish-brown

(sometimes gray) on a dark base

• 2-5++cm

For in vitro diagnostic use only. This product should be used only by adequately trained personnel with knowledge of microbiological techniques in the laboratory.

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Page 6

ORGANISM

PHYSIOLOGY

◆

Precision Test Strip

Available

GROWTH

COLONY

LaMotte BioPaddles™ TECH DOCUMENT

Call: 800-344-3100

Email: tech@lamotte.com

SAB

IMAGE

Epicoccum spp.

Fusarium spp.

• Catalase (+) ◆

• Ascomycete

• Catalase (+) ◆

• Ascomycete

+++

• Wooly / Cottony / Felty

• yellow, orange, red, brown

pigment

• 2-5++cm

• Wooly / Flat (sometimes

mucous-like)

• White, yellow, pink, purple, or

pale brown pigment

• 1-2cm

Microsporum spp.

• Catalase (+) ◆

• Ascomycete

Muccor spp.

• Catalase (+) ◆

• Zygomycete

+++

For in vitro diagnostic use only. This product should be used only by adequately trained personnel with knowledge of microbiological techniques in the laboratory.

• Glaborous (smooth) / Downy /

Wooly / Powdery

• White at first, later becoming grayish yellow to blue green with age

• 1-9+cm

• Wooly, Fluffy (like cotton

candy)

• White at first, later becoming grayish yellow to blue green with age

• 2-5++cm

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ORGANISM

PHYSIOLOGY

◆

Precision Test Strip

Available

GROWTH

COLONY

LaMotte BioPaddles™ TECH DOCUMENT

Call: 800-344-3100

Email: tech@lamotte.com

SAB

IMAGE

Penicillium chrysogenum (notatum)

Penicillium roqueforti

Penicillium digittum

• Catalase (+) ◆

• Ascomycete

• Catalase (+) ◆

• Ascomycete

• Catalase (+) ◆

• Ascomycete

+++

 Granular, velvet-

like/powdery, flat

• Initially white, then various shades of green blue-green or yellow-green pigment

• 2-5++cm

• Granular, dull green in color

arachnoid (with many spiderweb-like fibers) colony margins)

• 2-5++cm

• Wooly, Fluffy (like cotton

candy)

• White at first, later becoming green with age

• 2-5++cm

Rhizopus spp.

• Catalase (+) ◆

• Zygomycete

+++

For in vitro diagnostic use only. This product should be used only by adequately trained personnel with knowledge of microbiological techniques in the laboratory.

• Cottony

• White to blackish grey (black

fruiting bodies)

• 5++mm (rapidly spreading)

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Page 8

ORGANISM

PHYSIOLOGY

◆

Precision Test Strip

Available

GROWTH

COLONY

LaMotte BioPaddles™ TECH DOCUMENT

Call: 800-344-3100

Email: tech@lamotte.com

SAB

IMAGE

Saccharomyces cerevisiae

Stachybotrys spp.

Torula spp.

Trichoderma spp.

• Catalase (+) ◆

• Ascomycete

• Catalase (+) ◆

• Ascomycete

• Catalase (+) ◆

• Ascomycete

• Catalase (+) ◆

• Ascomycete

+++

• Creamy white to tannishcream

• Circular, entire, raised to convex, w/ glistening surface

• 1-4mm

• Dark gray

• Powdery

• white, pink, orange, black

pigment

• 2-5++cm

•Arrowhead / Circle or Heart

shape

• Grey, white to brown pigment

with age

• 2-5++cm

•

Cottony

•

White / later scattered green

or yellow-green patches (rings)

•

2-5++cm

IMAGE PENDING

•

Trichophyton spp.

• Catalase (+) ◆

• Ascomycete

Wooly with indented boarders

•

White to brownish-tan pigment

•

2-5++cm

For in vitro diagnostic use only. This product should be used only by adequately trained personnel with knowledge of microbiological techniques in the laboratory.

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ENUMERATION

SAB

Very Light Light Moderate

TVC/TCC

LaMotte BioPaddles™ TECH DOCUMENT

Call: 800-344-3100

Email: tech@lamotte.com

TVC/TCC (Total Viable Count/ Total Colony Counts)

Colony Counts <1000

Count colonies

TVC/TCC Count = Count x 2.5

Colony Counts >1000

Use chart

TVC/TCC Count = Count x 2.5

(Based on a 40 mL sample)

Example: Inoculated NUT/TTC paddle showing approximately 1000 CFU/100 mL.

For in vitro diagnostic use only. This product should be used only by adequately trained personnel with knowledge of microbiological techniques in the laboratory.

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Page 10

LaMotte BioPaddles™ TECH DOCUMENT

Call: 800-344-3100

Email: tech@lamotte.com

DISPOSAL

Twist to remove paddle from vial. Fill vial to 40 mL fill line with 1:9 dilution of household bleach (5.25% sodium hypochlorite). Replace paddle in vial. Allow 15 minute contact time. Remove paddle. Discard bleach solution. Replace paddle in vial and dispose. Alternatively, loosen cap and microwave for 30 seconds, autoclave, or incinerate.

GLOSSARY:

Catalase Test Catalase enzyme will react with hydrogen peroxide to produce oxygen if the bacteria is

catalase positive.

Lactose Test Lactose positive bacteria can ferment available lactose in the agar producing an acid which

lowers the pH. Lactose negative bacteria are non-fermenting.

Indole Test Biochemical test to determine the ability of an organism to split indole from the amino acid

tryptophan. P. vulgaris is indole positive while P. mirabilis is indole negative.

Oxidase Test Oxidase positive bacteria contain cytochrome c oxidase which will turn an indicator dark blue. In

contact with oxidase negative bacteria, the indicator will remain colorless.

Urease Test Bacteria containing urease will hydrolyze urea to ammonia and carbon dioxide

causing an alkaline environment which changes the color of a pH indicator from yellow to fuchsia.

β-D-Glucoronidase The presence of E. coli is determined when both β-D-Glucoronidase and Indole

Reaction are positive, and the organism is gram negative.

Gram Staining A method for differentiating bacteria into two groups – gram positive and gram negative –

based on the chemical and physical properties of their cell walls. Often the first step in identifying bacteria.

For in vitro diagnostic use only. This product should be used only by adequately trained personnel with knowledge of microbiological techniques in the laboratory.

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LaMotte POL-H User Manual

Specifications and Main Features

Frequently Asked Questions

User Manual