1. Presents
Overview Part 1 MultiChrome

2. Veterinary Microbiology Culture System
MultiChrome TM
Veterinary Microbiology Culture System
Procedures and Techniques For Performing
In-house Cultures using Kacey MultiChrome™ Bi-pates.
Gram Positive
Gram Negative
Exp 3/10
Lot #123
MultiChrome GP(-)
MultiChrome GP(+)

3. Sensi-Rings™ Quick Reference Card
Kacey Diagnostics Quick Reference For Culture & Sensitivity Study 1 2 3
INNOCULATION OF CULTURE BI-PLATE
1) Remove the plate from refrigerator & pre-heat in the degrees for minutes prior to inoculating the Kacey MultiChrome bi-plate.
2) Using a sterile inoculating loop ( 20uL) for liquids and a Kacey Sterile Swab for solids ,inoculate the sample onto both sides of the Kacey Multichrome
Bi-plate, by utilizing a zigzag streaking motion.
3) Replace clear lid and place Kacey MultiChrome
Bi-plate in the incubator upside down (inverted position) and incubate at 37°C +/- 2 degree C for no less then 24 hours . The plate should be examined after 24 hours, but no later than 48 hours after incubation.
TRANSFER TO MULLER HINTON PLATE USING KACEY “WST” TUBES
1) Taking a Kacey Sterile Swab or Loop carefully dab into the three (3) different places containing the bacteria on the MultiChrome Bi-plate.
2) Immediately place the sterile Kacey Swab or Loop containing the bacteria into the Kacey Working Solution tube (WST which contains 1.0 ml of sterile 0.085%solution). Mix the Kacey Swab or Loop with a gentle twirling motion while in the WST tube for approximately 3-5 seconds.
3) Compare sample to the Kacey Turbidity Standard (white cap) KTS. Sample should match in turbidity, if not add more WST sol (green top) or more sample to match KTS tube. (see enclosed Turbidity Card).
INOCULATION OF MULLER HINTON (MH) PLATE
1) Using the Kacey Swab or loop containing the now diluted WST & Turbidity adjusted dilution streak the 1st time the entire periphery of the Mueller Hinton plate making a 360° circle.
2) Streak a 2nd time again the Muller Hinton Plate with a fresh sample of the WST dilution usingbroad strokes starting at theto 3 o’clockposition.
3) Streak a 3rd time again the Muller Hinton plate with a fresh sample of the WST dilution using wide broad strokes starting at the 11 to 5 o’clock position
4) Remove “Sensi-Ring” from the foil pouch with tweezers at the inner tab & place the “Sensi-Ring” facing down onto the MH plate tapping down in non-disk areas, label the specimen to be incubated.
5) Place the MH Inoculated plate into an incubator upside down, set timer & incubate for 24 hours. Remove & read inhibition zones with Kacey Clear Acetate Reader. GN GP
24 Hrs GN GP 1 2 3
New Kacey Sterile Swab
Sample
KTS
White Tube
WST
Dilution
24 Hrs
Rotate Swab 3-5 sec
Compare to KTS tube
(white top) & match turbidity by adding more WST(grn top) solution or more sample
Quick reference only. Refer to detailed instruction manual for additional information.

4. BACTERIA AS A PATHOGEN
In veterinary medicine we routinely see them affecting nine areas:
Outer Ear Infections
Urinary Tract Infections
Conjunctival infections
Genital infections
Wound/abscess
Skin Infections
Upper Respiratory Infections
Mastitis
Septicemia

5. MultiChrome™ Supplies For Each Test
1 MultiChrome™ bi-plate
A sterile “Rayon”* swab included or sterile loop
included with MultiChrome™ kit.
Incubator: We recommend the Kacey Mini-Incubator
with its individual timers for up to six cultures.
MultiChrome GP(+)
Exp 3/10
Lot #123
MultiChrome GP(-)
*Always use Rayon swabs as chemicals in cotton
can alter results.

6. Kacey Mini Incubator Up Close
Small foot print (8”w x 13”hx12”d)
Precise Digital temperature control
Numbered trays fit both round & rectangle plates
Six individual timers
Audible/Visible alerts
Energy efficient, UL listed
Made in the USA
1year warranty
Patent Pending
MultiChrome GP(+)
Exp 3/10
Lot #123
MultiChrome GP(-)
Just Load… Press Timer...and Walk Away!

7. Pre-heat incubator. Pre heat incubator (98.6 F or 37 C default) which
will take approximately 30 minutes. The Kacey
Mini Incubator will automatically default to this
temperature when plugged in.
98.6 F or 37C

8. Pre-heat MultiChrome™ plate.
Remove a plate from the refrigerator and thaw
MultiChrome™ plate for approx 9-10 min. Place plate
upside down to avoid condensation landing on media.
The Kacey Mini Incubator has a “Red” pre-heat timer
button which allows you set a 10 minute warning.
Exp 3/10
Lot #123
MultiChrome GP(+)
MultiChrome GN(-)

9. Obtain sample and inoculate
MultiChrome™ plate.
Using a sterile loop or rayon swab obtain a sample
from the affected area.
Streak both the Gram (+) and (-) chambers of the
plate in a zigzag fashion.
Exp 3/10
Lot #123
MultiChrome GP(+)
MultiChrome GN(-)

10. Incubate sample. Cover MultiChrome™ plate with lid and turn upside
down.
Place in incubator and incubate for 24 hours. On the
Kacey Mini Incubator press one of six pre-set 24
hours timers and walk away.
24 hours
Exp 3/10
Lot #123
MultiChrome GP(+)
MultiChrome GN(-)

11. Read sample. At 24 hours remove from incubator and read.
Note whether the colony is a Gram (+) or (-) as
you reference the following color chart. Also, keep
in mind the types of bacteria most associated with
different affected areas of the body (see the next slide).
24 hours

12. Commonly found bacteria
Based on results, a further course of action can
now be with taken to determine the most efficacious
antibiotic(s) using the Sensi-Ring™ sensitivity system.
The following is a list of commonly found Aerobic
Bacteria in small animal practice:
Gram Positive: Staphylococci, Streptococci, Enterococci
Gram Negative: E coli, Proteus, Klebsiella, Pseudomonas
Ear: Staphylococci, Pseudomonas, Proteus, E. coli, Enterococci, Streptococci
UTI: Staphylococci, Proteus, E. coli, Klebsiella, Enterococci, occasionally Pseudomonas
Skin & Wound: Staphylococci, Streptococci, Pseudomonas, Enterococci, Proteus

13. Color reference chart
Gram Positive
Strep (+)
Mixed Staph/Strep (+)
This is a partial list of the various bacteria seen on MultiChrome™. Some bacteria may look slightly different than shown, but the colors and Gram (+), (-) designation remain constant. This library will continue to grow for use a reference guide.
Gram Negative

14. Bacterial pathogen probability charts for Canine and Feline:
These reference charts suggest the highest and lowest percentage for infection
of various parts of the body by common pathogenic bacteria. Once identified
proceed with accepted veterinary antibiotic protocols for treatment of pathogens.
Species: Canine
Percentage
Probability
Bacteria
UTI
Ear
Bone
Skin
Genital
Wound
Blood
URI
Staph Intermedius
10-15%
25-30%
40-50%
60-70%
15-25%
25-35%
30-35%
E. Coli
>50%
10-20%
20-30%
35-45%
15-20%
Enterococcus faecalis
<10%
NGS
Proteus mirabilis
15%
20-25%
Obligate anerobes
Pseudomonas
Klebsiella
20-35%
Pasturella
10-25%
NGS= Not Generally Seen
Species: Feline
Percent
Probability
Bacteria
Conjunctival
Genital
UTI
Wound
URI
Skin
Pasturella
10-20%
15-25%
10-15%
30-40%
>50%
E. Coli
NGS
30-35%
40-45%
15-20%
20-30%
Proteus mirabilis
<10%
Pseudomonas
Obligate anerobes
10-25%
25-35%
Klebsiella
Chlamydia psittoci
50-75%
B-hemolytic strep
NGS= Not Generally Seen
Probability reference charts taken from “Target: The Antimicrobial Reference Guide to Effective Treatment” , second edition, revised.
Author: Dr. David Aucoin, DVM, Diplomate of the American College of Veterinary Clinical Pharmacology. Data on file, Kacey® Asheville, N.C

15. Treatment. Many organisms can be easily identified and treated
after the use of the MultiChrome™ system without
further testing. However, for organisms that have
proven resistant to typical antibiotic regimens it may
be necessary to conduct a Sensitivity test.
Kacey Diagnostics has made “Sensitivity” testing easy,
accurate and cost effective as the MultiChrome™ by
introducing the Sensi-Ring system in the next section.

16. Sensi-Rings™ Quick Reference Card
Kacey Diagnostics Quick Reference For Culture & Sensitivity Study 1 2 3
INNOCULATION OF CULTURE BI-PLATE
1) Remove the plate from refrigerator & pre-heat in the degrees for minutes prior to inoculating the Kacey MultiChrome bi-plate.
2) Using a sterile inoculating loop ( 20uL) for liquids and a Kacey Sterile Swab for solids ,inoculate the sample onto both sides of the Kacey Multichrome
Bi-plate, by utilizing a zigzag streaking motion.
3) Replace clear lid and place Kacey MultiChrome
Bi-plate in the incubator upside down (inverted position) and incubate at 37°C +/- 2 degree C for no less then 24 hours . The plate should be examined after 24 hours, but no later than 48 hours after incubation.
TRANSFER TO MULLER HINTON PLATE USING KACEY “WST” TUBES
1) Taking a Kacey Sterile Swab or Loop carefully dab into the three (3) different places containing the bacteria on the MultiChrome Bi-plate.
2) Immediately place the sterile Kacey Swab or Loop containing the bacteria into the Kacey Working Solution tube (WST which contains 1.0 ml of sterile 0.085%solution). Mix the Kacey Swab or Loop with a gentle twirling motion while in the WST tube for approximately 3-5 seconds.
3) Compare sample to the Kacey Turbidity Standard (white cap) KTS. Sample should match in turbidity, if not add more WST sol (green top) or more sample to match KTS tube. (see enclosed Turbidity Card).
INOCULATION OF MULLER HINTON (MH) PLATE
1) Using the Kacey Swab or loop containing the now diluted WST & Turbidity adjusted dilution streak the 1st time the entire periphery of the Mueller Hinton plate making a 360° circle.
2) Streak a 2nd time again the Muller Hinton Plate with a fresh sample of the WST dilution usingbroad strokes starting at theto 3 o’clockposition.
3) Streak a 3rd time again the Muller Hinton plate with a fresh sample of the WST dilution using wide broad strokes starting at the 11 to 5 o’clock position
4) Remove “Sensi-Ring” from the foil pouch with tweezers at the inner tab & place the “Sensi-Ring” facing down onto the MH plate tapping down in non-disk areas, label the specimen to be incubated.
5) Place the MH Inoculated plate into an incubator upside down, set timer & incubate for 24 hours. Remove & read inhibition zones with Kacey Clear Acetate Reader. GN GP
24 Hrs GN GP 1 2 3
New Kacey Sterile Swab
Sample
KTS
White Tube
WST
Dilution
24 Hrs
Rotate Swab 3-5 sec
Compare to KTS tube
(white top) & match turbidity by adding more WST(grn top) solution or more sample
Quick reference only. Refer to detailed instruction manual for additional information.

17. Step 1: Obtaining live cultured bacteria from a Kacey MultiChrome™ bi-plate.
Using the yellow loop, take 3 or 4 passes through
the bacterial growth on the MultiChrome™.
For bacterial colonies in both chambers see next slide.

18. Sensi-Ring™ Supplies For Each Test
Kacey Mueller Hinton Plate
Loop
Rayon Swab
Red Top Turbidity Tube
Appropriate Sensi-Ring™
Kacey Micro Incubator
For added convenience we also offer a Culture & Sensitivity Starter Kit
enough for four complete tests with all the supplies you need. Product #20241

19. Procedure For MultiChrome bi-plates with bacteria on both the Gram Positive and Negative side.
In this case, you will prepare two Kacey Maxi Mueller
Hinton plates as indicated. One for the (+) and one for
the (-).
To each MH plate you will add the appropriate Sensi-Ring
disk Example: place a Gram (+) in the MH plate with Gram (+) and
Gram (-) in the MH plate with Gram (-). Then incubate each.

20. Mixing with Saline (WST) to dilute
Insert the yellow loop into the saline and twirl
to make a cloudy solution. This is an important
step to avoid overgrowth of a bacterial colony.

21. Comparing Sample to Standard
Compare your red top tube against the Standard (white top tube) which comes which every kit. They should match closely in turbidity. You can place the barcode card behind them to compare. If sample is too opaque, add more WST solution from dropper enclosed.
Standard
White Tube
Sample
Place Behind Tubes
Kacey C&S Turbidity Card
Directions On Back

22. Sensi-Rings™ Quick Reference Card
Kacey Diagnostics Quick Reference For Culture & Sensitivity Study 1 2 3
INNOCULATION OF CULTURE BI-PLATE
1) Remove the plate from refrigerator & pre-heat in the degrees for minutes prior to inoculating the Kacey MultiChrome bi-plate.
2) Using a sterile inoculating loop ( 20uL) for liquids and a Kacey Sterile Swab for solids ,inoculate the sample onto both sides of the Kacey Multichrome
Bi-plate, by utilizing a zigzag streaking motion.
3) Replace clear lid and place Kacey MultiChrome
Bi-plate in the incubator upside down (inverted position) and incubate at 37°C +/- 2 degree C for no less then 24 hours . The plate should be examined after 24 hours, but no later than 48 hours after incubation.
TRANSFER TO MULLER HINTON PLATE USING KACEY “WST” TUBES
1) Taking a Kacey Sterile Swab or Loop carefully dab into the three (3) different places containing the bacteria on the MultiChrome Bi-plate.
2) Immediately place the sterile Kacey Swab or Loop containing the bacteria into the Kacey Working Solution tube (WST which contains 1.0 ml of sterile 0.085%solution). Mix the Kacey Swab or Loop with a gentle twirling motion while in the WST tube for approximately 3-5 seconds.
3) Compare sample to the Kacey Turbidity Standard (white cap) KTS. Sample should match in turbidity, if not add more WST sol (green top) or more sample to match KTS tube. (see enclosed Turbidity Card).
INOCULATION OF MULLER HINTON (MH) PLATE
1) Using the Kacey Swab or loop containing the now diluted WST & Turbidity adjusted dilution streak the 1st time the entire periphery of the Mueller Hinton plate making a 360° circle.
2) Streak a 2nd time again the Muller Hinton Plate with a fresh sample of the WST dilution usingbroad strokes starting at theto 3 o’clockposition.
3) Streak a 3rd time again the Muller Hinton plate with a fresh sample of the WST dilution using wide broad strokes starting at the 11 to 5 o’clock position
4) Remove “Sensi-Ring” from the foil pouch with tweezers at the inner tab & place the “Sensi-Ring” facing down onto the MH plate tapping down in non-disk areas, label the specimen to be incubated.
5) Place the MH Inoculated plate into an incubator upside down, set timer & incubate for 24 hours. Remove & read inhibition zones with Kacey Clear Acetate Reader. GN GP
24 Hrs GN GP 1 2 3
New Kacey Sterile Swab
Sample
KTS
White Tube
WST
Dilution
24 Hrs
Rotate Swab 3-5 sec
Compare to KTS tube
(white top) & match turbidity by adding more WST(grn top) solution or more sample
Quick reference only. Refer to detailed instruction manual for additional information.

23. Why use Kacey Sensi-Ring™?
In house rapid identification (24 hours) of bacterial antibiotic
sensitivity using 8 of the most common veterinary antibiotic
choices per ring.
Cost Effective: Less than 1/3 the cost of a commercial laboratory
like Antech and Idexx.
Specific Sensi-Ring for both Gram Positive and Gram Negative
System specific Sensi-Ring:
Urinary Tract Infection
- Ear Infection
Skin and Wound Infection

24. Removing the diluted sample with enclosed Rayon* Swab
Take a sample using the applicator tip of the swab.
* Do not use cotton swabs even if sterile. Cotton will release
a chemical that will alter the results.

25. Inoculate diluted bacteria onto Kacey Maxi* Mueller Hinton plate
Using the diluted bacteria solution on the soaked
swab, evenly distribute the sample onto a warmed
Kacey Maxi Muller Hinton plate (37 degrees min).
Streak the plate as shown
below , 1-7, 9-3 o’clock
*Kacey Maxi Mueller Hinton plates are “true” 100mm plates. Most
commercially sold Muller Hinton plates are 88mm and will not work
with this system.

26. Apply the Sensi-Ring™
Pick the appropriate Sensi-Ring™ ie. Gram Positive,
Gram Negative, UTI or Skin/wound depending on
samples origin.
Grasp the tab on the inside of the ring and lower
onto the agar upside down with lettering facing agar,
centering the ring with tweezers.

27. Tamp down Sensi-Ring™ Make sure that all of the disks touch the agar
plate to allow antibiotic diffusion.

28. Incubate Upside Down Place Kacey Maxi Mueller Hinton plate now
inoculated upside down to prevent any contamination.
Place into a preheated Kacey incubator at 37 degrees
for 24 hours.
24 hours

29. Remove and read after 24 hours
Carefully remove plate from incubator keeping it
upside down so that Sensi-Ring letters can be seen.

30. Use the Kacey Zone Reader Card
Place Kacey Maxi Mueller Hinton Plate upside down
with antibiotic letters facing up.
Place Zone Reader over the plate and align the
notch of the Reader to the notch of the Sensi-Ring.
Concentric circles should now encircle all of the disks.
Read the Inhibition Zone in mm for each disk.
Notch

31. Log Test Results Using the Patient Lab Report form supplied log
inhibition zones for each antibiotic disk.

32. Treat with appropriate antibiotic
Based on results, a course of action can now be
taken with the most efficacious antibiotic(s).
The following is a list of commonly found Aerobic
Bacteria in small animal practice:
Gram Positive: Staphylococci, Streptococci, Enterococci
Gram Negative: E coli, Proteus, Klebsiella, Pseudomonas
Ear: Staphylococci, Pseudomonas, Proteus, E. coli, Enterococci, Streptococci
UTI: Staphylococci, Proteus, E. coli, Klebsiella, Enterococci, occasionally Pseudomonas
Skin & Wound: Staphylococci, Streptococci, Pseudomonas, Enterococci, Proteus

33. MRSA & MRSP
Can be detected with Sensi-Ring™ Skin and Wound

34. Sensi-Rings™ case samples
Case study: 2 year old canine which cultured on the MultiChrome™ both a large colony of Gram Positive Staph in pink and a smaller Strep colony in blue. The colonies when tested for sensitivity using the Sensi-Ring™ indicated sensitivity to a variety of antibiotics not previously used.
Staph
Strep

35. UTI and Ear samples

36. Sensi-Rings™ Ability to identify different strains of bacteria.
Sensi-Rings™ used to identify antibiotic sensitivity
on different strains of cultured Staphylococci.

37. Disposal of plate & clean up
Once read, plates should be sprayed with a bactericidal
agent, covered with its lid taped shut, placed in a plastic
bag and disposed of in your bio-hazard bin.
If you have a Kacey Sanitizer unit (coming soon) follow
its instructions. Place lid back on the plate, tape it shut, bag
it and dispose of it in your bio-hazard bin.
Dispose of all used supplies in your bio-hazard bin.
Spray your counters down and wash your hands in an
aseptic manor. Remember, these are live cultures.

38. “Kacey Products Providing Better Animal Care”
1854 A Hendersonville Rd
Asheville, NC 28803
Ph: 1(828)
Fax: 1(828)
“Kacey Products Providing Better Animal Care”
Kacey Diagnostics © 2014 All Rights Reserved
Rev
“MultiChrome”, “Sensi-Rings” are trademarks of Kacey® Inc.