63
Journal ofFood Protection. Vol. 45, No. I, Pages 63· 731January 1982)
Copyright©, International Association of Milk, Food, and Environmental Sanitarians
Microbial Flora of Livers, Kidneys and Hearts ·from Beef, Pork
and Lamb: Effects of Refrigeration, Freezing and Thawing
M. 0. HANNA, G. C. SMITH, J. W. SAVELL, F. K. McKEITH and C. VANDERZANT"'
Department ofAnimal Science, Texas Agricultural Experiment Station, TexasA&M University, College Station, Texas 77843
(Received for publication February 24, 1981)
ABSTRACT
Aerobic plate counts (APC) of livers, kidneys and hearts
obtained from beef, pork and Iamb soon after slaughter were
nearly always <104 and often <103 per cm 2 • Differences in APC
of different sites of the same liver, kidney or heart, within each
species, were not significant (P>0.05). APC of livers, kidneys
and hearts from pork and Iamb after storage for 1, 3 or 5 days
at 2 C were not significantly different (P>0.05) from those at
day 0. APC of beef livers, kidneys and hearts after 5 days at 2 C
differed significantly (P<0.05) from those at day 0, 1 and 3.
Temperature abuse of fresh organs for 6-12 h at 30 C before
freezing caused major increases in count. Frozen storage of
livers, kidneys and hearts (4 days at -20 C) did not cause
significant changes in APC. The initial microbial flora of fresh
livers, kidneys and hearts was varied with coryneform bacteria
and Micrococcus sp. often constituting a major part (>25%) of
the microbial flora. After storage for 5 days at 2 C,
Pseudomonas sp. more often became a major part of the
microbial flora of liver samples. Frozen storage for 4 days at
-20 C did not change the microbial flora of beef samples
greatly; in pork and lamb, coryneform bacteria more frequently
became a major part of the microbial flora after freezing.
Changes in pH of livers, kidneys and hearts during storage for 5
days at 2 C were small.
Numerous reports are available in the research
literature which describe the numbers and types of
bacteria on fresh meats. Most of these reports deal with
the microbial flora of surface and interior parts of
muscles of beef, pork and lamb carcasses, only a few
reports relate to the microbial flora of organs, such as
livers, kidneys and hearts (1,3,4). Gardner (1) reported
that the initial aerobic plate counts (APC) of fresh pork
liver was approximately 104 10 5 per cm 2 • The initial
surface microbial flora was varied and consisted of
Flavobacterium, Alcaligenes, Micrococcus, Leuconostoc,
Acinetobacter sp., Microbacterium thermosphactum,
coryneform bacteria and lactic streptococci. Storage of
pork live_rs for 7 days at 5 C in air resulted in a
proteolytic-type spoilage, with APC of 108 - 10 9 per cm 2•
Spoilage was caused by Pseudomonas, Alcaligenes,
JOURNAL OF FOOD PROTECTION, VOL. 45. JANUARY 1982
Escherichia sp., M. thermosphactum and lactic strepto-
cocci.
According to Shelef (4), initial counts of beef livers
from supermarkets were approximately 10 5 per g. The
microbial flora consisted of gram-positive cocci, spore-
formers, coliform bacteria and gram-negative rods.
Following storage in air for 7 - 10 days at 5 C, the livers
became organoleptically unacceptable due to souring, at
which time counts had reached 7-8 x 107 per g. Lactic
acid bacteria were predominant in the spoiled livers.
Patterson and Gibbs (3) reported on the microbial
flora of edible offal (liver, kidney, heart, tongue,
diaphragm). The initial microbial flora was varied, with
counts ranging from 104 - 10 5 per cm 2 • Shelf-life of the
products in air was 7 - 10 days at 4 C; when stored in
vacuum packages at 1 C for 3 weeks, products had an
additional shelf-life in air of 3 - 4 days at 4 C.
The microbiological quality of organ meats is
important to the United States meat industry since large
quantities, primarily beef livers, are shipped in the frozen
state to Europe for subsequent retail display and sale or
for manufacturing purposes. Previous reports (1,3) have
shown that the microbiological quality of organ meats
often is inferior because of poor sanitation during
handling and inadequate chilling and/or freezing before
shipment. The present study reports on the effects of
refrigeration, freezing and thawing on the microbial flora
and pH oflivers, kidneys and hearts from beef, pork and
lamb. A companion report (2) presents information on
the effect of vacuum-packaging vs. packaging in
polyvinyl chloride on the microflora of beef livers, beef
kidneys and pork livers.
MATERIALS AND METHODS
Samples
Livers, kidneys and hearts from beef, pork and lamb were obtained
from several meat packaging plants in Texas. Samples were
transported to the laboratory in polyvinyl chloride (PVC) film, covered
with ice. Refrigerated samples were kept in PVC film at 2 ± 1 C.
Samples to be stored frozen were wrapped in aluminum foil and frozen
in a freezer at -20 C. Before sampling, they were thawed overnight at
2C.
64 MICROBIAL FLORA OF LIVERS, KIDNEYS AND HEARTS

For the experiments in which pork livers and kidneys were frozen
immediately after slaughtering and dressing and after storage for 6 or
12 hat 30 C, organs were taken from hogs that were processed at the
Texas A&M University Meats Laboratory under identical conditions
over a period of a few hours. In the experiments on the effect of
defrosting on the microbial flora of beef livers and kidneys,
commercially processed livers and kidneys were used. Beef livers, two
per box, were wrapped individually in unsealed polyethylene film.
Kidneys were packaged in 10-lb. boxes. When samples were subjected
to various treatments (storage time, before and after freezing), each
sample was cut with a sterile knife into as many parts as were required
for the various treatments.
Ana~vses
Sampling was conducted by removing 10-cm2 areas (2mm thick)
from the surface (usually from 1-3 sites) of the organs with sterile
scalpels. Each sample was placed in 100 ml of sterile 0.1 o/o peptone in a
Stomacher bag and was macerated for 1 min in a Stomacher-400. In
experiments comparing counts of different sites of an organ, samples
from different sites were placed in separate Stomacher bags, in all
others they were combined before maceration. One-tenth-ml volumes
of appropriate dilutions were plated on prepoured plates of tryptic soy
agar (TSA) by the spread-plate method. Plates were incubated for 48 h
at 25 C. A few of each of the colony types appearing on each countable
plate were picked and placed on TSA slants. Slants were incubated for
48 h at 25 C. Identity of isolates was determined by biochemical tests
and identification schemes previously described by V anderzant and
Nickelson (6). Each of the colony types, now identified to the generic or
species level, then was expressed as a percentage of the total number of
colonies appearing on the countable plate. The pH value of samples
was measured after blending a 3-g sample with 25 ml of 0.005 M
iodoacetate for 1 min in a blender. The pH was measured with a
Coming Model 12 pH meter. Statistical analyses of the bacteriological
count data were made by using two-way analysis ofvariance (5).
RESULTS
Log aerobic plate counts (APC) of beef livers, kidneys
and hearts ranged from 2.00 to 2.81, from 2.52 to 3.00
and from 1.70 to 2.99, respectively (Table 1). The pH
values of beef livers, kidneys and hearts ranged from 6.18
to 6.29, from 6.36 to 6.62 and from 6.17 to 6.60,
respectively. Log APC of pork livers, kidneys and hearts
ranged from 2.30 to 3.92, from 3.04 to 4.75 and from 1.48
to 4.04, respectively. The pH values of pork livers,
kidneys and hearts ranged from 6.42 to 6.48, from 6.72 to
7.00 and from 6.05 to 6.55, respectively. Log APC of
lamb livers, kidneys and hearts ranged from 2.15 to 5.32,
from 3.08 to 3.46 and from 2.59 to 3.63, respectively. The
pH values oflamb livers, kidneys and hearts ranged from
6.15 to 6.39, from 6.48 to 6.68 and from 5.65 to 5.67,
respectively. Differences in APC of different sites of the
same liver, kidney or heart, within each of the species,
were not significant (P>0.05).
Log APC of beef livers, kidneys and hearts after
storage under refrigeration for 5 days at 2 C ranged from
2.36 to 4.08 (Table 2). Compared with counts at day 0,
APC of beef livers, kidneys and hearts following storage

TABLE 1. Aerobic plate counts ofthree different sites and pH value of livers, kidneys and hearts.

Aerobic plate count 0og10 /cm2)

Sample - No. Site 1 Site 2 Site 3 pH
Beef
-1
Liver 2.57 2.00 2.81 6.29
-2 2.53 2.60 2.45 6.20
-3 2.48 2.30 2.72 6.18
Kidney- 1 3.00 3.00 2.80 6.41
-2 3.00 2.65 2.62 6.62
-3 2.52 2.57 2.74 6.36
Heart - 1 2.18 1.70 2.23 6.60
2 2.11 2.66 2.74 6.17
-3 2.04 1.90 2.99 6.48
Pork
Liver -1 2.30 2.42 2.46 6.42
.2 3.92 3.81 3.52 6.48
3 3.76 3.45 3.84
Kidney- 1 3.38 3.04 3.38 7.00
-2 4.75 4.38 4.38 6.72
-3 3.73 3.84 3.34
Heart - 1 1.70 2.11 1.48 6.55
-2 3.71 3.32 3.53 6.05
3 4.04 3.43 4.00
Lamb
Liver -1 4.85 4.94 5.32 6.39
-2 2.15 3.53 3.48 6.15
-3 4.67 4.23 4.98 6.19
Kidney- 1 3.30 3.30 6.48
-2 3.08 3.46 6.68
-3 3.23 3.34 6.49
Heart . 1 3.63 3.59 3.15 5.67
-2 2.95 2.97 2.59 5.65
-3 3.04 2.83 2.74 5.65

JOURNAL OF FOOD PROTECTION. VOL. 45, JANUARY 1982

 HANNAETAL. 65
TABLE 2. Effect ofrefrigerated storage at 2 Con aerobic plate count and pH oflivers, kidneys and hearts.

Aerobic plate count (log10 /cm 2) at day

Sample No. 0 3 5 0 1 3 5
Beef
Liver .1 2.38 2.66 2.30 2.79 6.13 6.05 6.10 5.82
-2 2.68 2.67 1.85 2.36 6.00 6.00 6.12 5.72
.3 2.38 2.62 2.76 2.64 6.12 6.10 6.15 6.08
Kidney -1 2.92 2.53 3.00 4.08 6.16 6.25 6.50 6.52
.2 2.56 3.04 2.70 2.76 6.35 6.32 6.70 6.50
-3 2.69 3.23 3.04 3.18 6.41 6.31 6.70 6.80
Heart . 1 2.08 2.08 2.57 3.04 6.60 6.58 6.90 6.72
.2 1.60 1.74 2.26 3.32 6.59 6.41 6.95 6.50
-3 2.53 2.48 2.70 3.40 6.21 6.12 6.42 6.32
Pork
Liver . 1 2.34 2.58 2.70 2.92 6.00 5.92 6.05 6.00
.2 1.70 2.48 2.92 2.40 6.05 5.95 6.05 5.95
.3 1.20 2.36 2.36 1.63 6.09 6.10 6.00 6.00
Kidney -1 2.68 2.38 2.60 2.20 6.75 6.70 6.85 6.62
-2 2.57 2.40 2.78 3.11 6.75 6.71 7.00 6.90
.3 2.80 2.61 2.60 2.51 6.75 6.66 7.00 6.72
Heart . 1 2.00 1.48 2.00 1.00 6.20 6.10 6.50 6.42
.2 2.00 1.78 2.00 2.15 6.50 6.51 6.75 6.58
.3 2.36 1.30 2.00 2.38 6.30 6.38 6.80 6.49
Lamb
Liver . 1 3.49 3.42 4.23 3.26 6.10 5.99 5.85 5.89
.2 3.36 3.20 3.23 4.40 6.10 5.88 5.55 5.99
.3 3.32 3.38 3.61 3.52 6.30 6.01 6.00 6.29
Kidney- 1 3.28 3.08 6.62 6.89
-2 3.34 3.60 6.51 6.99
.3 3.20 3.85 6.51 6.99
Heart . 1 3.08 3.70 3.81 5.59 5.58 5.90
.2 3.04 3.08 3.15 5.90 5.96 6.61
.3 3.11 3.97 3.32 6.13 6.21 6.51

for 5 days at 2 C increased in 8 of 9 comparisons.
Increases in log APC ranged from 0.20 to 1.72. Only
counts at day 5 were significantly (P<0.05) higher than
those at 0, 1 or 3 days. Decreases in pH of beef livers
during refrigerated storage (5 days at 2 C) ranged from
0.04 to 0.31, increases in pH of beef kidneys during the
same period ranged from 0.15 to 0.39. Refrigerated
storage of beef hearts caused only minor changes (0.09 to
0.12) in pH.
Log APC of pork livers, kidneys and hearts after
storage under refrigeration for 5 days at 2 C ranged from
1.00 to 3.11 (Table 2). Compared with counts at day 0,
APC of pork livers, kidneys and hearts following storage
for 5 days at 2 C increased in 6 of 9 comparisons.
Increases in log APC from day 0 to day 5 for pork livers,
kidneys and hearts were relatively small and ranged from
0.02 to 0.70. Counts of samples after 1, 3, 5 days of
storage were not significantly (P>0.05) different from
those obtained at day 0. Changes in pH of pork livers,
kidneys and hearts during refrigerated storage were
small {0 to 0.22); decreases {0.03 to 0.13) were noted in 4
of 9 samples and increases {0.08 to 0.22) were observed in
4 of 9 samples.
Log AP.C of lamb livers, kidneys and hearts following
storage for 5 days at 2 C ranged from 3.08 to 4.40
(Table 2). Compared with counts at day 0, APC of lamb
livers, kidneys and hearts following refrigerated storage
for 5 days increased in 7 of 9 comparisons. Increases in
log APC during 5 days of storage were relatively small
and ranged from 0.11 to 1.04, decreases in log APC
ranged from 0.20 to 0.23. Counts obtained after 1, 3 and
5 days of storage were not significantly (P>0.05) different
from those at day 0. During refrigerated storage for 5
days, minor decreases (0.01 to 0.21) in pH of lamb livers
occurred. During the same period, pH values of lamb
kidneys and hearts increased slightly, ranges of increases
in pH were 0.27 to0.48 and 0.31 to 0.71, respectively.
APC (Table 3) of livers, kidneys and hearts from beef,
pork and lamb before and after freezing for 4 days at
-20 C were not significantly different (P>0.05).
The microbial flora of beef livers (Table 4) at day 0
consisted of Micrococcus, Streptococcus, Staphylo·
coccus, coryneform bacteria, Pseudomonas, and Morax·
ella·Acinetobacter sp. Micrococcus sp., coagulase nega-
tive staphylococci and coryneform bacteria contributed a
major part (>25o/q) of the microbial flora of one or more
of the beef livers at day 0. The microbial flora of beef
livers following storage at 2 C for 5 days was different
from that at day 0; changes in microbial flora, however,
were not consistent. After refrigerated storage of beef
livers for 5 days, Streptococcus spp. were dominant in
sample 1, yeasts and coryneform bacteria in sample 2,

JOURNAL OF FOOD PROTECTION, VOL. 45, JANUARY 1982

66 MICROBIAL FLORA OF LIVERS, KIDNEYS AND HEARTS

TABLE 3. Effect offrozen storage at -20 Con aerobic plate Staphylococcus and Moraxella-Acinetobacter sp. All of
count oflivers, kidneys and hearts. these, except for Moraxella-Acinetobacter sp., consti-
tuted a major part (>25 o/o) of the microbial flora of one or
Aerobic plate count (log10/ cm2)
more of the three pork livers at day 0. Following
Sample No. Before freezing After freezing (4d) refrigerated storage of pork livers for 5 days, the
Beef microbial flora of sample 1 was dominated by
Liver -1 2.32 2.73 Staphylococcus sp., that of sample 2 by Moraxella-
-2 2.08 2.04 Acinetobacter sp. and that of sample 3 by Micrococcus
-3 2.18 3.08 and Streptococcus sp.
Kidney- 1 2.52 2.53
The microbial flora of pork kidneys (Table 5) at day 0
-2 2.46 3.43
-3 2.95 2.94
consisted of Micrococcus, Streptococcus. Staphylo-
Heart • 1 3.04 2.83 coccus, coryneform bacteria, and Flavobacterium sp.
.2 3.11 2.23 Micrococcus, Staphylococcus sp. and coryneform bac-
.3 3.30 2.34 teria constituted a major part (>25%) of the microbial
Pork flora of one or more of the pork kidneys at day 0.
Liver 1 3.61 3.26 Following storage of pork kidneys for 5 days at 2 C, the
.2 3.68 3.60 microbial flora of sample 1 was dominated by
.3 3.18 3.57 coryneform and Micrococcus sp., that of sample 2 by
Kidney· 1 3.69 3.20 Pseudomonas, Micrococcus, and Staphylococcus sp.,
.2 3.74 3.57 that of sample 3 by Staphylococcus sp. and coryneform
-3 3.52 3.54
Heart 1 3.23
bacteria.
3.49
-2 2.93 2.56
The microbial flora of pork hearts at day 0 (Table 5)
.3 2.65 2.81 consisted of Micrococcus, Streptococcus, Staphylo-
Lamb coccus, coryneform bacteria and Pseudomonas sp. All of
Liver . 1 2.62 3.08 these, except for Staphylococcus sp., constituted a major
.2 2.91 3.08 part (>25 %) of the microbial flora of one or more of the
-3 2.45 2.73 pork hearts at day 0. After storage of pork hearts for 5
Kidney 1 2.90 2.72 days at 2 C, the microbial flora of samples 1 and 2 was
-2 2.70 2.93 dominated by Micrococcus sp., that of sample 3 by
.3 2.58 2.72 Pseudomonas sp.
Heart 1 1.85 2.32
.2
The microbial flora of lamb livers at day 0 (Table 6)
1.48 1.48
.3 consisted of Micrococcus, Staphylococcus, coryneform
1.70 2.00
bacteria. Pseudomonas, Moraxella-Acinetobacter, Fla-
and Pseudomonas spp. in sample 3. vobacterium sp. and E. coli. Micrococcus, coryneform
bacteria. Pseudomonas and Moraxella-Acinetobacter sp.
The microbial flora of beef kidneys (Table 4) at day 0
made up a major part (>25%) of the microbial flora of at
consisted of Micrococcus, Streptococcus, Staphylo·
coccus, coryneform bacteria, Pseudomonas, Moraxella·
least one of the lamb livers at day 0. Following storage of
Acinetobacter and Flavobacterium sp. Micrococcus,
lamb livers for 5 days at 2 C, the microbial flora of
sample 1 was dominated by coryneform bacteria, that of
Streptococcus, coryneform bacteria and Flavobacterium
sp. made up a major part (>25%) of the microbial flora of sample 2 by Staphylococcus and Micrococcus sp., that of
sample 3 by coryneform bacteria and Streptococcus sp.
one or more of the beef kidneys at day 0. Changes in the
microbial flora of beef kidneys during refrigerated The microbial flora of lamb kidneys at day 0 (Table 6)
storage for 5 days at 2 C were not consistent; in samples 1 consisted of Micrococcus, Staphylococcus, coryneform
and 2, Streptococcus sp. became dominant; in sample 3 bacteria and yeasts. Coryneform bacteria and Micro·
coccus sp. were a dominant part of the microbial flora at
the microflora consisted of coryneform bacteria and
Micrococcus sp.
day 0. Following storage for 5 days at 2 C, coryneform
The microbial flora of beef hearts (Table 4) at day 0 bacteria constituted from 58.2 to 72.5% of the microbial
consisted of Micrococcus, Staphylococcus, coryneform flora oflamb kidneys.
bacteria and M oraxella·Acinetobacter sp. Micrococcus The microbial flora of lamb hearts at day 0 (Table 6)
and/or coryneform bacteria dominated the microbial consisted of Micrococcus, Streptococcus. Staphylo-
flora of all samples at day 0. The microbial flora of beef coccus, coryneform bacteria, Moraxella·Acinetobacter,
hearts was not as varied as those of beef livers or beef Flavobacterium sp. and molds. Coryneform bacteria,
kidneys. Following storage of beef hearts for 5 days at Micrococcus and Streptococcus sp. made up a major
2 C, the microbial flora of samples 1 and 3 was part (>25 o/o) of the microbial flora of one or more of the
dominated by Pseudomonas sp., that of sample 2 by lamb hearts at day 0. After storage for 5 days at 2 C,
Micrococcus sp. coryneform bacteria and Micrococcus sp. made up over
The microbial flora of pork livers (Table 5) at day 0 60%ofthe microbial flora of lamb hearts.
consisted of Micrococcus, Streptococcus, Pediococcus, The microbial flora of beef livers. kidneys and hearts,

JOURNAL OF FOOD PROTECTION, VOL.45, JANUARY 1982

 TABLE 4. Distribution of microbial flora of beef livers, kidneys and hearts stored for 0 to 5 days at 2 C.

Percentage of microbial populationa

Storage time
-No. Mic Cor Ps M-A Flav H.a. y M Uk
Liver- l 0 47.9 16.9 18.3 11.3 5.6
3 28.6 60.3 11.1
5 17.0 69.2 9.3 2.2 2.2
C3 Liver· 2 0 22.9 3.5 27.1 46.5
§a
3 19.0 66.7 9.5 4.8
~ 5 28.6 14.3 57.1
~ Liver· 3 0 47.9 46.6 5.5
3 63.6 8.1 13.9 6.9 7.5
~
0 5 19.5 5.3 75.2
0 40.5
Kidney 1 0 23.8 2.4 13.1 16.7 3.6
~ 3 17.3 3.8 3.8 75.0 ::t:
0
t;j 5 0.8 95.2 2.4 1.6 z>
(J_ z
Kidney 2 0 27.8 41.7 5.6 25.0 >
:::1 18.0 tTl
,_,
0 3 68.0 14.0
~ 5 3.4 67.2 29.3 >
<: r
0 Kidney 3 0 16.3 34.7 10.2 6.1 32.7
r
.j>, 3 26.4 6.6 18.9 37.7 10.4
·"'
..... 5 46.2 53.8
z> Heart- 1 0 20.8 8.3 54.2 16.7
c::: 3 9.5 58.1 32.4
>::0 5 5.7 14.4 3.8 76.1
....:
..... Heart 2 0 75.0 25.0
'-0
00
N 3 69.4 8.3 22.2
5 87.8 12.2
Heart 3 0 85.3 14.7
3 37.4 29.3 27.3 6.1
5 4.1 95.9
aMic Micrococcus, Strep =Streptococcus, Staph- =coagulase-Staphylococcus, Cor= Coryneform bacteria, Ps =Pseudomonas,
M-A Moraxella·Acinetobacter, Flav Flavobacterium, H.a. =H. alvei, Y =Yeasts, M =Molds, Uk Unknown.

-..:1
"'
 0\
00

TABLE 5. Distribution of microbialflora ofpork livers, kidneys and hearts stored for up to 5

Cl Storage time ~
~ ·No. Mic Ped M-A Flav M R
::0
Liver- 1 0 27.3 54.4 12.1 6.1 0
~ txl
0 5 24.0 52.0 16.0 8.0
"'1 r>
Liver- 2 0 93.3 6.7
~ 5 14.9 2.7 14.9 58.1 9.4
0 5
::0
tl Liver- 3 0 4.1 44.9 36.7 14.3 >
;; 5 46.1 38.5 7.7 7.7 0
0 'Tl
Kidney- 1 0 68.8 31.2 t::
t;i <
(') 5 31.3 50.0 18.8
::;j Kidney- 2 0 29.7 59.5 10.8
!;;]
0 .!"'
:<: 5 28.4 28.3 1.5 40.3 1.5 ~
< ...,
0 Kidney- 3 0 22.2 11.1 39.7 27.0
r-' 5 18.7 43.7 25.1 12.5 z0
-!>- !;1
.!"' Heart- 1 0 100.0 V>
.... 5 100.0
z> z>
e Heart- 2 0 30.0 10.0 60.0 0
> 5 75.0 18.7 6.3 ::c
::0
-< Heart- 3 0 39.1 52.2 ~
.... 8.7 ::0
...,
'1:>
00 5 4.2 4.2 91.6 V>

"" Micrococcus, Strep =Streptococcus, Ped = Pediococcus, Staph-= coagulase-Staphylococcus, Cor= Corynefonn bacteria,
Pseudomonas, M-A = Moraxella·Acinetobacter, Flav Flavobacterium, M = Molds.
 TABLE 6. Distribution of microbial flora oflamb livers, kidneys and hearts stored forO to 5 days at 2 C.

Percentage of microbial populationa

Storage time
Sample No. (days) Mic Strep Staph· Cor Ps M-A Flav E.e. y M Uk
Liver- 1 0 9.8 6.6 52.5 31.1
C5 3 9.1 9.1 36.4 15.1 27.3 3.0
~ 5 17.6 61.2 10.5 10.8
Liver- 2 0 44.4 28.9 20.0 2.2 4.4
~ 3 53.3 26.7 20.0
c
"tl 5 32.0 50.0 18.0
;:s Liver- 3 0 4.8 42.9 52.4
g 3 29.3 54.9 3.6 12.2
~ 5 3.0 31.8 36.4 19.7 9.1 ~
c Kidney- 1 0 15.8 5.7 78.5
t;j z>
<"l 5 17.6 24.2 58.2 z
>
::.:l Kidney- 2 0 9.9 4.9 75.8 9.4
c ~
~ 5 7.5 72.5 20.0 >
< Kidney- 3 0 60.4 6.1 33.5 r
0
r 5 4.5 61.2 34.2
~
!J' Heart - 1 0 10.6 5.7 74.8 8.9
>
-z 3 30.0 12.0 52.0 6.0
c: 5 40.0 4.6 30.8 13.8 4.6 6.2
>
::>:1
Heart- 2 0 3.1 27.1 44.8 17.7 5.2 2.1
-< 3 21.3 12.3 54.9 3.3 8.2
~ 5 10.3 11.0 53.7 25.0
tv
- Heart- 3 0 27.3 1.6 54.7 12.5 3.9
3 25.5 27.7 31.9 6.4 8.5
5 42.1 7.6 4.8 26.3 5.3 13.9
aMic =Micrococcus, Strep Streptococcus, Staph- coagulase-Staphylococcus, Cor = Coryneform bacteria, Ps = Pseudomonas,
M-A Moraxella·Acinetobacter, Flav =Flavobacterium, E.c. =E. coli, Y =Yeasts, M =Molds, Uk =Unknown.

0\
10
70 MICROBIAL FLORA OF LIVERS, KIDNEYS AND HEARTS

used in trials on the effect of freezing on the microbial
flora of variety meats, consisted primarily of coryneform
bacteria and/or Micrococcus sp. (Table 7). After freezing
(day 4), coryneform bacteria and/or Micrococcus sp.
constituted over 50% of the microbial flora in 7 of the 9
samples. In some instances, changes in the microbial
flora because of freezing were consistent. For example,
freezing caused decreases in the percentages of
coryneform bacteria in livers; in beef hearts freezing
caused increases in the percentage of coryneform
bacteria and decreases in percentage of Micrococcus sp.
Coryneform bacteria, and to a lesser extent Micro-
coccus, Flavobacterium and Staphylococcus sp., were a
major part of the microbial flora of pork livers, kidneys
and hearts used in experiments on the effect of freezing
on the microbial flora of these variety meats (Table 8).
After freezing, coryneform bacteria with (4 samples) or
without (4 samples) Micrococcus sp. comprised over 50%
of the microbial flora of 8 of the 9 samples.
Micrococcus, Staphylococcus, Streptococcus, Mor-
axella-Acinetobacter, Flavobacterium and coryneform
bacteria were a major part (>2So/o) of the microbial flora

TABLE 7. Distribution of microbial flora of beef livers, kidneys and hearts before and after frozen storage.

Storage time Percentage of microbial population a

Sample- No. (days) Mic Strep Staph- Cor M-A Flav E.c. S.i. y

Liver - 1 0 25.8 38.7 17.7 17.7

4 47.8 36.0 14.9 1.2
Liver- 2 0 14.3 85.7
4 9.4 59.4 31.2
Liver - 3 0 27.3 25.0 11.4 36.4
4 44.4 19.4 19.4 16.7
Kidney -1 0 10.6 24.2 54.5 10.6
4 28.4 62.7 7.5 1.5
Kidney- 2 0 45.6 7.0 17.5 15.8 14.0
4 9.3 90.7
Kidney- 3 0 44.4 44.4 11.1
4 43.5 4.3 4.3 8.7 39.1
Heart- 1 0 81.7 1.6 16.7
4 51.7 7.3 34.1 2.9 3.9
Heart- 2 0 100.0
4 51.0 5.9 23.5 11.8 7.8
Heart 3 0 78.8 6.1 15.1
4 40.9 13.6 43.9 1.5
aMic =Micrococcus, Strep Streptococcus, Staph- =coagulase-Staphylococcus, Cor Coryneform bacteria, M-A = Moraxella·
Acinetobacter, Flav =Flavobacterium, E.c. E. coli, S.!. S.liquefaciens, Y =Yeasts.

TABLE 8. Distribution of microbialflora ofpork livers, kidneys and hearts before and after frozen storage.

Storage time Percentage of microbial population a

Sample No. (days) Mic Strep Staph- Cor Ps M-A Flav E.h. y
Liver 1 0 27.4 14.5 46.0 2.4 6.5 3.2
4 7.6 11.3 39.6 7.6 9.4 24.5
Liver- 2 0 21.1 2.4 12.5 35.9 28.1
4 1.7 74.2 6.7 13.3 4.2
Liver- 3 0 9.1 22.7 22.7 4.6 38.6 2.3
4 18.9 59.5 9.0 9.9 2.7
Kidney- 1 0 30.6 6.1 26.5 8.2 8.2 20.4
4 18.4 7.4 12.3 47.2 4.9 9.8
Kidney- 2 0 75.5 11.3 5.7 7.5
4 27.0 10.8 27.0 13.5 21.6
Kidney· 3 0 15.1 30.3 36.4 18.2
4 45.7 8.6 14.3 28.6 2.9
Heart- 1 0 11.1 4.7 55.6 13.4 15.2
4 36.7 20.0 13.3 30.0
Heart· 2 0 11.8 20.0 32.9 8.2 27.1
4 8.3 11.1 55.6 11.1 11.1 2.8
Heart· 3 0 22.3 51.1 13.3 13.3
4 15.4 7.7 72.3 1.5 3.1
aMic Micrococcus, Strep Streptococcus, Staph·= coagulase-Staphylococcus, Cor= Coryneform bacteria, Ps =Pseudomonas,
M-A Moraxella·Acinetobacter, Flav Flavobacterium, E.h. =E. herbicola, Y =Yeasts.

JOURNAL OF FOOD PROTECTION, VOL. 45. JANUARY 1982

 HANNAETAL. 71

of one or more of the nine lamb livers, kidneys and hearts
used in experiments on the effect of freezing on the
microbial flora of these variety meats (Table 9).
Micrococcus sp. constituted 25% or more of the
microbial flora of 5 of the 9 samples at day 0. After
freezing of lamb livers, coryneform bacteria with or
without Streptococcus and Micrococcus sp. were
dominant. After freezing of lamb kidneys, Micrococcus
sp. with either coryneform bacteria, Moraxella-Acineto-
bacter spp. or yeasts were dominant. The microbial flora
oflamb hearts after freezing consisted primarily of either
Staphylococcus or M oraxella-Acinetobacter sp.
Experiments on simulated temperature abuse of pork
livers and kidneys showed (Table 10) that counts of fresh
livers after holding at 30 C for 6 or 12 h increased by 1.07
and 2.99 logs, those for fresh kidneys by 1.38 and 3.08
logs, respectively. When livers that had been held frozen
for 1 month were compared with counterpart livers that
had been held at 30 C for 30 min, 6 h or 12 h, but never
frozen, differences in count between livers were -0.14,
-0.69 and +0.06log, respectively; comparable differences
in count for kidneys were 0.18, 0.90 and 0.78 log,
respectively. The initial microbial flora of pork livers and
kidneys was varied but Micrococcus sp. clearly were
dominant (73.5-98.3o/o). Holding fresh livers and kidneys
for 12 hat 30 C caused changes (>10%) in the microbial
flora which included a decrease in Micrococcus sp. and
an increase in homofermentative lactobacilli; in kidneys
held for 12 h at 30 C there was also an increase in
coagulase-negative Staphylococcus and in Acinetobacter
sp. Storage of fresh liver for 6 h at 30 C did not cause
changes in the microbial flora exceeding 10%; in kidneys
Micrococcus sp. decreased and homofermentative
lactobacilli increased. In the livers and kidneys stored for
1 month at -20 C, Micrococcus sp. and/or coryneform
bacteria were dominant.
When boxes of commercially frozen kidneys and livers
were set at room temperature (25 C) for 24 or 48 h,
increases in count of kidneys and livers after 24 h were
0.26 and 0.46 log and after 48 h were 2.82 and 3.76logs,
respectively (Table 11).
DISCUSSION
Initial APC of livers, kidneys and hearts in this study
(Tables 1,2,3) were considerably lower than those
reported by others (1,3,4). However, the initial counts of
commercially frozen variety meats in cardboard boxes
were similar to the counts reported by Gardner (1),
Patterson and Gibbs (3) and Shelef (4). Except for the
commercially frozen variety meats, livers, kidneys and
hearts used in this study were obtained soon after
slaughter to determine what could be achieved
microbiologically with good handling and storage
practices.
Results indicate that counts of different sites of organs
were not significantly different. This is important,
because when samples were subjected to different
treatments (time of storage, before and after freezing),
each sample was subdivided into as many parts as were
required for the various treatments. If organs are
promptly and properly refrigerated, no major increases
in microbial count occurred over a 5-day period
(Table 2). When, however, they are subjected to
temperature abuse for 6-12 h at 30 C before freezing
(Table 10), major increases in count occurred. In
addition, defrosting of frozen items for 24-48 h at 25 C or
break-down of freezing temperature maintenance may
lead to increases in microbial count (Table 11). These

TABLE 9. Distribution of microbial flora of lamb livers, kidneys and hearts before and after frozen storage.

Storage time of microbial

Sample· No. (days) Mic Strep Staph· Cor Ps M-A Flav y
Liver· 1 0 35.7 42.1 11.1 7.1 4.0
4 22.2 13.9 52.8 11.1
Liver· 2 0 12.2 8.2 74.3 5.3
4 8.3 16.7 72.2 2.8
Liver· 3 0 27.8 7.2 4.8 1.2 59.0
4 36.8 33.7 27.7 1.8
Kidney· 1 0 56.3 28.7 5.0 10.0
4 47.2 7.5 26.4 17.0 1.9
Kidney- 2 0 10.0 18.0 38.0 8.0 6.0 20.0
4 31.8 12.9 31.8 23.5
Kidney· 3 0 10.5 18.4 15.8 21.1 34.2
4 26.4 5.7 3.8 20.7 11.3 32.1
Heart- 1 0 42.8 14.3 14.3 28.6
4 9.5 52.4 9.5 28.6
Heart· 2 0 66.7 33.3
4 100.0
Heart- 3 0 25.0 75.0
4 60.0 20.0 20.0
aMic =Micrococcus, Strep =Streptococcus, Staph-= coagulase-Staphylococcus, Cor= Coryneform bacteria, Ps- Pseudomonas,
M-A = Moraxella·Acinetobacter, Flav =Flavobacterium, Y =Yeasts.

JOURNAL OF FOOD PROTECTION, VOL. 45, JANUARY 1982

 -..J
N

TABLE 10. Microbial}lora ofpork livers and kidneys immediately after slaughter-dressing [0 storage time], after storage at 30 C
for 30 min, 6 or 12 h and after frozen [-20 C]jor 1 month.

a Storage I..ogAPC Hom. 3:

~ time cm 2 Bac Cor Lac Mic M.t. + Ac C.d. E.h. Flav Ps y (")
'""'
~
Liver, frozen Oh 2.79 0.4 97.4 0.5 1.9 0
~ l:tJ
0..., after 12 h 12 h 5.78 7.1 6.7 16.4 54.9 0.9 6.7 7.5 >
..., t"'
at30 C lmonth 5.72 2.6 6.6 62.1 17.8 3.9 1.7 1.1 0.6 3.8 "'i
0 t"'
Kidney, frozen Oh 3.45 2.7 0.9 85.8 0.9 6.1 3.7 0
g after 12 h 12 h 6.53 4.4 6.6 11.6 42.1 19.5 1.1 11.0 2.9 1.1 ~
>
~ at 30 C 1month 5.75 0.4 3.8 71.0 0.8 12.4 8.2 3.6 0
0 "'i
Liver, frozen Oh 2.08 1.7 90.6 3.4 1.7 2.8 t"'
....
t;j
\'l after 6 h 6h 3.15 0.8 5.9 89.4 2.0 2.0 <
:::! [i;J
0 at 30 C 1month 3.84 64.4 1.0 18.5 3.8 0.3 8.1 3.1 1.0 Y'
;<!: Kidney. frozen Oh 3.28 0.4 1.2 98.3 0.2 ~
< after6 h 6h 4.66 0.7 30.2 64.6 4.0 0.6
0
t"'
.j:..
at30 C 1month 3.76 27.5 6.6 43.8 8.3 12.3 1.7 ~t"r1
Y' ....::
Liver, frozen Oh 2.64 25.0 73.5 1.6 til
....
after 30 min 1month 2.78 36.8 0.7 50.8 2.0 2.5 1.3 0.6 1.6 3.9 z>
z> 0
e at 30 C
> :I:
~ Kidney, frozen Oh 3.38 0.8 4.7 16.9 75.5 0.6 1.6 t"r1
....:: >
after 30 min lmonth 3.20 12.5 4.0 61.7 8.6 8.5 4.9
~ at 30 C ::1
Vl
N

a sac Bacillus, Cor = Coryneform bacteria, Hom. Lac Homofermentative Lactobacillus, Mic =Micrococcus, M.t. =Micro·
bacterium thermosphactum, Staph- =coagulase-Staphylococcus, Staph+= coagulase+Staphylococcus, Strep =Streptococcus,
Ac = Acinetobacter, Ale Alcaligenes, C.d. = Citrobacter diversus, E.cl. = Enterobacter cloacae, E.c. = Escherichia coli,
E.h. = Erwinia herbicola, Flav =Flavobacterium, Ps =Pseudomonas, Y Yeasts.
 HANNAETAL.
TABLE 11. Aerobic plate count of frozen beef kidneys and
livers before and after defrosting at room temperature [25 C]
or48 h.
Aerobic plate count Oog10 /cm2 )a at
Sample Oh 24 h 48 h
Kidney 5.85 6.11 8.67
Liver 5.00 5.46 8.76
a Average of2 samples.
results are of significance to meat packers who might
wish to sell variety meats in the domestic trade (not
usually frozen) and to meat packers who freeze variety
meats for subsequent transport-distribution to foreign
markets. Rapid chilling and/or freezing retards micro-
bial growth; maintenance of cold (for unfrozen product)
and frozen (previously frozen product) conditions also
retards microbial growth.
It is well known that freezing can cause sublethal
injury and death to many microbial species in foods.
Freezing of livers, kidneys and hearts did not cause
significant changes in APC. In general, changes in pH of
livers, kidneys and hearts during storage for S days at 2 C
were minor, which is in agreement with the lack of major
microbial proliferation during this period. The initial
microbial flora of livers, kidneys and hearts was varied,
similar results have been reported by Gardner (J), Shelef
(4) and Patterson and Gibbs (3). Coryneform bacteria
and Micrococcus sp. were frequently encountered and
constituted 25 o/o or more of the microbial flora of 17 of 27
and 12 of 27 samples, respectively. Pseudomonas sp.
made up 25 o/o of the microbial flora of only 2 of 27
samples. Although storage for S days at 2 C did not cause
major increases in counts, the number of samples on
which Pseudomonas sp. constituted 25o/o or more of the
microbial flora had increased from 2 of 27, to 7 of 27
JOURNAL OF FOOD PROTECTION, VOL. 45, JANUARY 1982
73
samples, among the heart samples alone they increased
from 1 of 9, to 4 of 9 samples. Gardner (J) encountered
Pseudomonas sp. as part of the aerobic spoilage flora of
pork liver. The microbial flora of livers, kidneys and
hearts before and after freezing was often dominated by
coryneform bacteria and Micrococcus sp. Before
freezing, Micrococcus sp. and coryneform bacteria
constituted 25 o/o or more of the microbial flora of 15 of 27
and 12 of 27 samples, respectively, after freezing of 14 of
27 and 17 of 27 samples, respectively. This more frequent
occurrence of coryneform bacteria at this level (>25o/o)
after freezing occurred in the pork and lamb samples
only. Experiments are now underway to determine the
effect of various commercial handling practices in meat
packing plants on the microbiological and organoleptic
qualities of variety meats.
ACKNOWLEDGMENTS
TA 16693 of the Texas Agricultural Experiment Station. This study
was supported in part by USDA-SEA Research Agreement Number
907-15·107.
REFERENCES
1. Gardner, G. A. 1971. A note on the aerobic microflora of fresh and
frozen porcine liver stored at 5• C. J. Food Technol. 6:225-231.
2. Hanna, M. 0., G. C. Smith, J. W. Savell, F. K. McKeith, and C.
Vanderzant. 1981. Effects of packaging methods on the microbial
flora of livers and kidneys from beef or pork. J. Food Prot.
3. Patterson, J. T., and P. A. Gibbs. 1979. Vacuum·packaging of
bovine edible offal. Meat Science. 3:209-222.
4. Shelef, L. A.l975. Microbial spoilageoffresh refrigerated beef liver.
J. Appl. Bacteriol. 39:273-280.
5. Snedecor, G. W. 1956. Statistical methods, 5th ed. The Iowa State
College Press, Ames, Iowa.
6. Vanderzant, C., and R. Nickelson. 1%9. A microbiological
examination of muscle tissue of beef, pork and lamb carcasses. J.
Milk Food Techno!. 32:357-361.