Simple Mnemonics for

Members of the
Enterobacteriaceae Using
Most medical students, clinical pathology resi-
Octal Numbers dents and medical technology students have, from
time to time, complained that they cannot re-
member for any long period of time the various
biochemical reactions upon which bacteriologists
base their classification of members of the En-
terobacteriaceae. Most pathology residents and
medical technology students learn these reactions
at the time of their rotation in microbiology, and
perhaps just at the time of their board examination,
only to be forgotten the next day. Some mnemonic
by James W. Lo, M.D., and Bernard R. Tess, Ph.D. codes are needed as an extra crutch by those
without a photographic memory or an extremely
logical mind. (The drawbacks of mnemonics are not
discussed here.)
We long have been looking for crutches to help
us and others to recall the more important
biochemical reactions for most of the bacteria of
clinical significance. The recently available En-
terotube 3 and API1 systems, as well as other compu-
ter programs,4,5'6 have aroused considerable in-
terest in those who realize the role that coding
systems and computers can play in the rapid
speciation of members of the Enterobacteriaceae
family. However, the logic involved may not easily
be understood by some bacteriology technologists
and there certainly are no simple mnemonic codes
for easy recall.
In the process of developing computerization of
bacteriology reports using the newly available
CLAS-800 (B-D Spear), we were introduced to the
octal and the binary numbers. It dawned upon us
that a simple mnemonic code could be constructed

Table 1
Octal Binary

0 0x22 + 0x21 + 0x2° = 0 0 0

1 0 x 2 2 + 0 x 2 l + 1 x2° = 0 0 1
2 0x22+1 x21 + 0x2° = 0 1 0
3 0 x 22 + 1 x 21 + 1 x 2° = 0 1 1
4 1x22 + 0 x 2 l + 0x2°=1 0 0
5 1 x 22 + 0 x 2 1 + 1 x 2° = 1 0 1
James W. Lo, M.D., and Bernard R. Tess, Ph.D., are 6 1 x 22 + 1 x 2 1 + 0 x 2° = 1 1 0
with the Microbiology Section, Pathology Depart- 7 1 x 22 + 1 x 21 + 1 x 2° = 1 1 1
ment, Mercy Hospital and Medical Center, Chicago.

24 LABORATORY MEDICINE • VOL 7, NO. 1, JANUARY 1976

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 for the biochemical results of most of the bacteria Table III
using the octal and the binary numbering systems.
TSI Group (Gr. 1) 1. Fermentation of lactose
The members of Enterobacteriaceae are chosen for
and sucrose
the purpose of illustration.
2. Gas production from
glucose
Method
3. Production of H2S
The mnemonics proposed for instant recall of the
results of the biochemical reactions of the mem- IMViC Group 1. Indole production
bers of Enterobacteriaceae is given in octal numbers (Gr. 2) 2 Methyl red reaction
(that is, based on powers of eight and using digits 3. Citrate utilization
0, 1, 2, 3, 4, 5, 6, and 7). The corresponding binary (Simmons)
numbers are as shown in table I.
Deaminase- 1. Phenylalanine
Each of the eight "octal numbers" corresponds to Decarboxylase deaminase
three bits of the binary numbers. If 0 of the binary Group (Gr. 3) 2. Lysine decarboxylase
number is chosen to represent a negative biochem- 3. Ornithine
ical reaction and 1 of the binary system to represent decarboxylase
a positive biochemical reaction, each bit of the
binary system can be used to characterize the result Miscellaneous 1. Urease
of a particular reaction. If three biochemical tests Group (Gr. 4) 2. Dulcitol
are grouped together in a previously assigned 3. Rhamnose
order, the results of these three tests can be
expressed by a set of three binary numbers or just
one octal number. tests is arbitrarily chosen and fixed as listed, there
is only one set of 12 results that is characterized by
A simple illustration will clarify this point. For an
an octal number of four digits. Hence, the number
example, with three successive bits of the binary
of 6631 (octal numbering system) means an or-
system denoting (a) fermentation of lactose and
ganism that ferments lactose and sucrose, pro-
sucrose, (b) production of gas from glucose and (c)
duces gas from glucose, H2S ( - ) ; Indole (+), MR
production of H2S, see table II.
(+), Citrate ( - ) ; Phenylalanine deaminase ( - ) ,
Lysine decarboxylase (+), Ornithine decarboxylase
Table II (+); Urease ( - ) , Dulcitol (-), and Rhamnose (+).
Biochemical Reaction Apparently, these biochemical reactions are the
characteristics of Escherichia species. Similar four-
Fermentation Gas digit octal numbers can be made for most members
of Lactose & from Binary Octal of the Enterobacteriaceae at the level of genera and
Sucrose Glucose H2S No. No. sometimes at the level of the species.

- - - 0 0 0 0 The mnemonic code applies only to the more

- - + 0 0 1 1 common members of the Enterobacteriaceae.
- - Hence, an organism has to be a Gram-negative rod
+ 0 1 0 2
that ferments glucose and is oxidase negative
- + + 0 1 1 3
- - before one can use the biochemical parameters for
+ 1 0 0 4
- genus or species determination.
+ + 1 0 1 5
+ + - 1 1 0 6 The first group of the biochemical reactions
+ + + 1 1 1 7 summarizes the more important information that
can be derived from the TSI agar or its equivalent,
Similar groups of three other biochemical reac- the second group of tests refers to the IMViC tests
tions are chosen and the results are again expressed with the VP reaction deleted because of spatial
in (a) binary and (b) octal numbers. In order to limitation, the third group of tests covers three
simplify the mnemonics, only 12 biochemical reac- important enzymes elaborated by certain bacilli of
tions (or four groups of reactions with three the Enterobacteriaceae, and the fourth group is a
reactions in each group) are chosen and assigned miscellaneous group with urease and two fermenta-
the order shown in table III. tion reactions.

Each group yields one octal number, and four The four octal numbers for these four groups
octal numbers in succession yield the results of 12 may be remembered as a set of four-digit numbers;
biochemical tests. If the order of the biochemical or one may prefer to commit to memory one, two

LABORATORY MEDICINE • VOL. 7, NO. 1, JANUARY 1976 25

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 or three octal numbers separately for a particular
organism depending on one's aptitude for num-
bers. This mnemonic system can be, therefore,
easily simplified or expanded at will. Also, the
mnemonic code does not commit one to the use of
a particular commercial kit (be it R/B improved, API
20 Enteric, Enterotube improved, Minitek, efc.) or
the conventional system.
It will be obvious from table IV, which lists the
binary numbers, octal numbers and the names of
the organisms (based on the CDC Enteric Bacteriol-
ogy Laboratory Chart2), that genus determination
can be obtained in most cases by knowing the first
three octal digits (Providencia and Arizona being
the exceptions). If all four digits are known, specia-
tion may be attempted in some cases (Table IV).

The important things to note are the following: Discussion

1. Remember the mnemonic code (in octal num- The preceding describes an attempt to produce a
bers) for the particular organism in question. simple mnemonic code for the purpose of rapid
2. Know how to convert octal numbers into recall of the more common biochemical results of
binary numbers. the major members of the Enterobacteriaceae fam-
3. Know the order of the 12 biochemical reac- ily. As such, it does not replace the detailed charts,
tions. profile registers and tables that are now available
for speciation purpose.
The whole idea is to present simple one to four
digit octal numbers that most of us, with practice,
Table IV can recall; the conversion from octal numbers back
Binary No. to binary numbers is known to most doctors and
Organism Gr 1 Gr. 3 Gr. 4
technologists and can be mastered within a short
Gr. 2 Octal No.
time by the uninitiated. Theoretically, one could
ESCHERICHEAE expand the number of biochemical parameters and
Escherichia sp. J_ 1° 1 1
9- 0 1» 1" 0 0 V 6 6 3 1 the corresponding binary and octal numbers to a
Shigella sp. 2' 0 0 0_ 1 0. 00 0» 0 0 0" 0 2 0 0 few dozen, but the simplicity of the mnemonics
EDWARDSIELLEAE
will be lost, and computers or large tables will be
necessary to give us instant recall of the test results
Edwardsiella _0 1 i_ ri o 0 1 1 0 £ 0_ 3 6 3 0
rather than the human brains.
SALMONELLEAE

Salmonella typhi _0 o 1 0 1 0 0 1 0_ 0 .0 0 12 2 0
The mnemonic code presented as such also does
not take into consideration the variable results of
Other Salmonella sp. 0_ 1 1_ 0 1-1 0 1 1 0 1 1 3 3 3 3
the biochemical reactions. This is a major limita-
Arizona Of 1 1 0 1 1 0 1 1 0 0 1 3 3 3 1
tion, but it could be circumvented if one cares to
Citrobacter sp. V 1 1_ 0 1 1 0 0_ 1" 1» 1" 1 3 3 17 remember more than one four-digit octal number
KLEBSIELLEAE for each genus. Maybe by increasing it to five
Klebsiella sp. 1 1 0 0£ 1 0 1 p_ 1 0 1 6 12 5 or six digits, a more comprehensive speciation can
Enterobacter aerogenes 1 1 0 0_0 1 0 1 1 _0 0 1 6 13 1
be done. The latter is up to the interested
microbiologist/technologist to try.
Enterobacter cloaceae 1 1 0 0_0 1 0 0 1 1 0 1 6 115

Enterobacter hafniae 0 1 0 0J3 0" 0 1 1 0 0 1 2 0 3 1 For others, it may turn out that a three-digit octal
Enterobacter number is all that is needed for their particular
agglomerans 0 1 0 0 0 1' 00 0 0 0 1 2 10 1 purpose (e.g., determination of the genus of a
Serratia sp. 1 1 0 0_0 1 0 1 1 0 0 j) 6 13 0 particular bacillus, given only a few parameters at
PROTEEAE the time of a microbiology practical examination).
Providencia 0 0 0 1 1 1 1 0 0 0 7 4 0
1° °. References
1
Proteus vulgaris 0 1 1 1 ° 2 oo 1 0 0 3 6 4 4
1. API Profile Recognition System (API Coder, API Profile Register, and
Proteus mirabilis 0 1 1 11 1 1 0 1 1 0 0 3 3 5 4 API Percent Chart), Analytab Products, Inc.

1 1 ° i° 1
Proteus morganii 1 0 0
2. Enteric Bacteriology Laboratory Chart: Differentiation of Enterobac-
0 1 0 2 6 5 4
teriaceae by Biochemical Tests. DHEW-PHS-CDC, Atlanta, GA. 30333,
Proteus rettgeri 0 0 0 1 1 1 i °1 1 0 0- 0 7 4 4 July 1973.
3. Enterotube Numerical Coding and Identification System for Enterobac-
Underlined binary numbers indicate important biochemical tests for teriaceae, 1973, Hoffmann-La Roche, Inc.
differentiation.
8 4. Friedman, R.B. ef a/.: Computer-assisted Identification of Bacteria, Am.
= Different biochemical types—regard as 1 if more than 50% are positive
J. Clin. Path. 60:395-403, 1973.
and 0 if more than 50% are negative.
b
= Shigella sonnei ferments lactose, sucrose and rhamnose slowly and 5. Ring, A.M.: Programs for the Programmable Calculator in the Labora-
decarboxylates ornithine. tory, Am. J. Clin. Path. 61:575-576, 1974.
c
= Lactose variable and sucrose negative. 6. Spencer, F., and Hyde, T.A.: An Approach to Microbiologic Diagnosis
d
= Both lactose and sucrose are variable. and Matrix Problems for the Small Hospital Laboratory Using a Small
Computer, Am. J. Clin Path. 60:264-267, 1973. [fl]

26 LABORATORY MEDICINE • VOL. 7, NO. 1, JANUARY 1976

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