Meat Science 97 (2014) 197206

Contents lists available at ScienceDirect

Meat Science
journal homepage: www.elsevier.com/locate/meatsci

Review

Functionality of liquid smoke as an all-natural antimicrobial in

food preservation
Jody M. Lingbeck a, Paola Cordero b, Corliss A. O'Bryan b, Michael G. Johnson b,
Steven C. Ricke a,b,c, Philip G. Crandall a,b,
a
b
c

Sea Star International LLC., 2138 East Revere Place, Fayetteville, AR 72701, USA
Department of Food Science and Center for Food Safety, University of Arkansas, 2650 Young Ave., Fayetteville, AR 72704, USA
Department of Poultry Science, Division of Agriculture, University of Arkansas, Fayetteville, AR 72704, USA

a r t i c l e

i n f o

Article history:
Received 16 October 2013
Received in revised form 28 January 2014
Accepted 2 February 2014
Available online 9 February 2014
Keywords:
Liquid smoke
Antimicrobial
Listeria monocytogenes
Salmonella

a b s t r a c t
The smoking of foods, especially meats, has been used as a preservation technique for centuries. Today, smoking
methods often involve the use of wood smoke condensates, commonly known as liquid smoke. Liquid smoke is
produced by condensing wood smoke created by the pyrolysis of sawdust or wood chips followed by removal of
the carcinogenic polyaromatic hydrocarbons. The main products of wood pyrolysis are phenols, carbonyls and
organic acids which are responsible for the avor, color and antimicrobial properties of liquid smoke. Several
common food-borne pathogens such as Listeria monocytogenes, Salmonella, pathogenic Escherichia coli and
Staphylococcus have shown sensitivity to liquid smoke in vitro and in food systems. Therefore liquid smoke has
potential for use as an all-natural antimicrobial in commercial applications where smoke avor is desired.
This review will cover the application and effectiveness of liquid smoke and fractions of liquid smoke as
an all-natural food preservative. This review will be valuable for the industrial and research communities in
the food science and technology areas.
2014 Published by Elsevier Ltd.

Contents
1.
2.
3.

Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Generation of liquid smoke from wood pyrolysis . . . . . . . . . . . .
Antimicrobial activity of liquid smoke . . . . . . . . . . . . . . . . .
3.1.
Possible mechanisms of antimicrobial action of liquid smokes . . .
3.2.
Activity of phenols . . . . . . . . . . . . . . . . . . . . . .
3.3.
Activity of carbonyls . . . . . . . . . . . . . . . . . . . . . .
4.
Antimicrobial activity of liquid smoke against Listeria . . . . . . . . . .
4.1.
In vitro effects on Listeria . . . . . . . . . . . . . . . . . . . .
4.2.
Antilisterial effects in ready-to-eat meats . . . . . . . . . . . .
4.3.
Genetic basis of the antimicrobial effects of liquid smoke on Listeria
5.
Effects of liquid smoke on Salmonella spp. . . . . . . . . . . . . . . .
6.
Effects of liquid smoke on E. coli . . . . . . . . . . . . . . . . . . . .
6.1.
In vitro effects of liquid smoke on E. coli . . . . . . . . . . . . .
6.2.
Effects of liquid smoke on E. coli in beef . . . . . . . . . . . . .
7.
Effect of liquid smoke on Staphylococcus . . . . . . . . . . . . . . . .
8.
Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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197
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205

1. Introduction
Corresponding author at: 2650 Young Ave., Fayetteville, AR 72704, USA.Tel.: +1 479
575 7686.
E-mail address: crandal@uark.edu (P.G. Crandall).

http://dx.doi.org/10.1016/j.meatsci.2014.02.003
0309-1740 2014 Published by Elsevier Ltd.

Traditional smoking of foods, especially meats, has been used as

a preservation technique for centuries. Wood smoke, in addition to

198

J.M. Lingbeck et al. / Meat Science 97 (2014) 197206

preserving food quality with its antioxidant and antimicrobial properties, also imparts a desirable color, avor and aroma to smoked foods.
Application of liquid smoke requires less time than traditional smoking,
is more environmentally friendly, and eliminates potentially toxic compounds while still imparting the desired avors and aromas of traditional smoking. Use of condensates or liquid smoke allows the processor
to control the concentration of smoke being applied more readily than
generating smoke by burning of wood (Suen, Fernandez-Galian, &
Aristimuo, 2001). Liquid smoke is traditionally applied to meat, sh
and poultry and it has also been used to impart avor to non-meat
items such as cheese, tofu and even pet food. Because the smoke avor
is concentrated, application of liquid smoke is best suited for use in marinades, sauces or brines or topically to processed meat items such as hot
dogs, sausage, ham and bacon (Rozum, 2009).
According to an annual poll conducted by The Center for Food Integrity consumers have less condence in the safety and quality of the food
supply and are demanding more all-natural and minimally processed
foods with less synthetic chemical additives (Andrews, 2012). Consumers also have increased interest in organic foods because they believe they are healthier, better tasting, or fresher than conventional
products (Wier & Calverley, 2002). However, although free of synthetic
chemicals, organic and all-natural foods are not exempt from bacterial
contamination and may require the addition of an all-natural antimicrobial to insure their safety. All-natural antimicrobials including those
derived from plants, animals and bacteria have been shown to be effective in increasing the safety of food products by destroying or limiting
the growth of bacterial pathogens. Several reviews have been written
on all-natural antimicrobials from bacterial, plant and animal origin
(Davidson, Critzer, & Taylor, 2013; Juneja, Dwivedi, & Yan, 2012; Rai &
Chikindas, 2011), as well as their use in organic poultry and meat production (Ricke, Van Loo, Johnson, & O'Bryan, 2012; Sirsat, Muthaiyan,
& Ricke, 2009). However, these reviews contain little or no information
on the use of liquid smoke as an effective all-natural antimicrobial. The
review by Holley and Patel (2005) provides a nice overview on the use
of liquid smoke as well as its antimicrobial properties in food systems,
especially in sh. This review builds on the information presented in
Holley and Patel (2005) as well as provides a more detailed and up to
date discussion on the effectiveness of liquid smoke as an all-natural
preservative in food products. We will examine the effectiveness of
liquid smoke, including ranges of microbial susceptibility and factors

affecting antimicrobial action and discuss currently understood mechanisms of action.

2. Generation of liquid smoke from wood pyrolysis
Liquid smoke is produced by condensing wood smoke created by the
controlled, minimal oxygen pyrolysis of sawdust or wood chips. The
wood is placed in large retorts where intense heat is applied, causing
the wood to smolder (not burn), releasing the gases seen in ordinary
smoke. These gases are quickly chilled in condensers, which liquees
the smoke. The liquid smoke is then forced through rening vats, and
then ltered to remove toxic and carcinogenic impurities. Finally, the
liquid is aged for mellowness. Fig. 1 shows a schematic of a typical liquid
smoke production facility. Factors inuencing the avor and antimicrobial properties of liquid smoke include the temperature of smoke generation, moisture content of the wood as well as the type of wood used to
generate the smoke (Simko, 2005). Common woods include hickory
and mesquite, but liquid smoke has also been prepared from rice hulls
(Kim et al., 2011, 2012), coconut shells (Zuraida, Sukarno, & Budijanto,
2011) and pecan shells (Van Loo, Babu, Crandall, & Ricke, 2012). In general, woods used to generate liquid smoke are roughly comprised of 25%
hemicellulose, 50% cellulose, and 25% lignins (Simko, 2005). See Table 1
for information about composition of specic woods. Pyrolysis occurs in
four stages starting with water evaporation, followed by decomposition
of hemicelluloses, cellulose decomposition and nally decomposition of
lignins. Pyrolysis of hemicellulose and cellulose occurs between 180 C
and 350 C and produces carboxylic acids and carbonyl compounds
while lignins are pyrolyzed between 300 C and 500 C and generate
phenols (Ramakrishnan & Moeller, 2002; Simko, 2005). Smoke avor
compounds, including phenols, are responsible for the smoke avor
and smoky aroma while carbonyl compounds impart a sweet aroma
and color to smoked meat products.
In addition to carbonyls, acids, and phenols, pyrolysis of wood often
generates unfavorable compounds such as polycyclic aromatic hydrocarbons (PAH). Polycyclic aromatic hydrocarbons are families of compounds, some which are naturally occurring, others are the result of
incomplete burning and are typically formed at pyrolysis temperatures
between 500 C and 900 C (Simko, 2005). The level of PAH formation is
also inuenced by the wood source (Guilln, Sopelana, & Partearroyo,
2000). Some PAH compounds such as benzo(a)pyrene (B(a)P), have

Fig. 1. Flow diagram of typical liquid smoke production.

J.M. Lingbeck et al. / Meat Science 97 (2014) 197206

Table 1
Wood carbohydrate composition (%).
Wood

Cellulose

Hemicellulose

Lignin

Apple
Cherry
Chestnut
Hard maple
Hickory
Mesquite
Red oak
White oak

20.7
20.7
21.4
17.2
41.4
8.0
58.6
21.4

6.9
3.4
3.6
17.2
1.7
8.0
3.4
3.6

37.9
13.8
32.1
55.2
24.1
44.0
24.1
39.3

From Chen and Maga (1993).

been shown to cause birth defects when pregnant mice were exposed to
more than 300 ppm in food. Foods containing levels greater than
900 ppm led to liver and blood defects in test animals (EPA, 2008).
The European Union (EU) regulations limit the amount of PAH allowed
in food while the US Food and Drug Administration has not set an upper
limit for PAH exposure (Dolan, Matulka, & Burdock, 2010). Because data
has shown that it is possible to reach lower levels of B(a)P in smoked
meats, acceptable levels of regulated PAH, specically B(a)P, will drop
from 0.005 to 0.002 ppm in 2014 for these foodstuffs sold in the EU
((EC) No. 835/2011). The 2014 level of PAH is set 150,000 times
below the levels known to cause birth defects. Although PAH are extremely toxic, they have low water solubility which allows liquid
smoke manufacturers to easily separate out these compounds from
their nished products using phase separation and ltration techniques.
For more information on PAH in liquid smoke see Guilln and Sopelana
(2003) and Simon, de la Calle, Palme, Meier, and Anklam (2005).
3. Antimicrobial activity of liquid smoke
Different woods generate different levels of phenols, carbonyls and
organic acids upon pyrolysis which affect their antimicrobial properties
(see Table 2). Liquid smokes from 20 different types of woods including
redwood, black walnut, birch, hickory, aspen, white oak, cherry
and chestnut were assessed for their antimicrobial properties against
Staphylococcus aureus and Aeromonas hydrophila in broth culture
(Boyle, Sofos, & Maga, 1988; Sofos, Maga, & Boyle, 1988). It was found
that wood smoke generated from Douglas r sapwood was inhibitory
to both the bacterial strains in that it delayed initiation of growth and
growth rates of these organisms while smoke from mesquite or lodge
pole pine did little to inhibit these pathogens (Boyle et al., 1988; Sofos
et al., 1988). In a separate study, liquid smoke from white mangrove,
mahogany and abura were shown to inhibit S. aureus and Escherichia
coli. Red mangrove and alstonia were able to inhibit S. aureus, but not
E. coli, indicating that not only does the type of wood affect the antimicrobial properties of liquid smoke but also that pathogenic organisms
have varying degrees of sensitivity to the ingredients of the liquid
smoke (Asita & Campbell, 1990). The susceptibility of major food
borne pathogens Listeria monocytogenes, Salmonella, E. coli and Staphylococcus to liquid smoke in laboratory media as well as in model meat systems will be reviewed. Table 3 summarizes the in vitro results of liquid
smoke against several food borne pathogens while Table 4 is a summary
of the antimicrobial activity of liquid smoke against pathogens in model
food systems.
3.1. Possible mechanisms of antimicrobial action of liquid smokes
Gram-positive and Gram-negative organisms may behave differently to exposure to liquid smoke or fractions of liquid smoke and there
may be varying susceptibility within differing strains of the same organism thus making it difcult to identify the mechanism and compounds
responsible for microbial inhibition (Sofos et al., 1988). The amount of
phenols present in liquid smoke condensates has been reported to
be approximately 9.911.1 mg/mL (Ramakrishnan & Moeller, 2002).

199

Phenolic compounds are known to disturb the cytoplasmic membranes

of bacteria and cause the intracellular uids to leak (Davidson, 1997).
Carbonyls have been reported in liquid smokes in amounts of approximately 2.6 to 4.6% (Milly, Toledo, & Ramakrishnan, 2005). The efcacy
of carbonyls as an antimicrobial can be inferred based on the 133 different aldehydes and ketones present in liquid smoke (Montazeri, Oliveira,
Himelbloom, Leigh, & Crapo, 2013). Carbonyls inhibit microbial growth
by penetrating the cell wall and inactivating enzymes located in the
cytoplasm and the cytoplasmic membrane (Milly, 2003). Carbonyls act
by condensing with the free, primary amino-groups in the polypeptide
chains, primarily in the side-chains of basic amino-acids. These aminogroups may be an essential part of active site of the enzyme, or they
may function to bind the substrate by hydrogen-bonding (Painter,
1998). Even if the carbonyls cannot access the interior of a microbial
cell, they can still inhibit growth by interfering with the uptake of nutrients. There are 3 proposed mechanisms involved in this interference,
termed A, B and C types.
Type A inhibition entails the sequestration of amino acids or ammonia
by condensation with the carbonyl compounds, thus lowering the effective concentration in the growth medium (Painter, 1998). Some carbonyls, including -keto-carboxylic acids, enediols, 3-hydroxyketones
and 1,3-diones, can also remove essential metal cations by chelation
(Painter, 1998). Type B inhibition is active against putrefactive bacteria
or molds which excrete exocellular proteases or glycanases to break
down proteins or glycans into small fragments which can be taken up
by the cells. Carbonyl compounds either inactivate the enzymes as
described for type A inhibition, or by immobilizing them in insoluble
particles or a three-dimensional polymeric network which physically
isolates them from their substrates (Painter, 1998). Type C inhibition
involves direct chemical modication of a substrate itself, so that it becomes less accessible or susceptible to the microbial enzymes (Painter,
1998). Antimicrobial activity of phenols and carbonyls is discussed
further in the following sections.

3.2. Activity of phenols

Early studies on antimicrobial activity of liquid smoke on
L. monocytogenes and other pathogens attributed the activity to phenolic compounds. Faith, Yousef, and Luchansky (1992) evaluated several
liquid smoke fractions and common smoke phenols for antilisterial activity. A three strain cocktail of L. monocytogenes including isolates
Scott A, V7 and 101 M was incubated in hot dog exudate containing
0.2% or 0.6% of the liquid smoke fraction CharSol Supreme (Red Arrow
Company, Manitowoc, WI) at 25 C for up to 114 h. L. monocytogenes
levels decreased in liquid smoke treated samples with D-values of 36
h for exudate containing 0.2% liquid smoke and 4.5 h for 0.6% treated
samples. When eleven individual smoke phenols were evaluated for
their antilisterial activity against L. monocytogenes Scott A in Tryptose
Broth, only isoeugenol was able to delay growth. The addition of acetic
acid further enhanced inactivation due to its bactericidal activity
(Young & Foegeding, 1993).
Suen (1998) measured the antimicrobial activity of seven different smoke fractions used in the Spanish food industry against
L. monocytogenes and other pathogenic microorganisms. The MIC values
were determined using an agar dilution technique with smoke extracts
prepared in buffer, pH adjusted to 7 and used at concentrations up to 2
the maximum level recommended by the manufacturer. Antimicrobial
activity was directly related to phenol concentrations in that extracts
possessing a high concentration of phenols (94 to 153 mg/kg)
corresponded to the fractions with the strongest antimicrobial properties. However, the fraction containing the highest concentration of
phenols was not the most active. The most active fraction, in addition
to having a high level of phenols (21 mg/kg), also contained the highest
concentration of acids (34 mg/kg) which may have contributed to the
antimicrobial action of this fraction.

200

J.M. Lingbeck et al. / Meat Science 97 (2014) 197206

Table 2
Chemical properties of commercial liquid smokes.
Liquid smoke tested

Manufacturer

pH

Titratable acidity as percent

acetic acid (wt/wt)

Phenol content
(mg/mL)

Carbonyl content
(g/100 mL)

References

Charsol Supreme

2.12.6

1416

1825

2025

Vitt et al. (2001)

2.22.8
No data
2.12.6
2.02.4
3.04.0
7.38.1
Not listed
2.53.3

1416
No data
10.512
1315
4.0 maximum
No data
1011
3.55.6

1523
2530
1015
914
3742
No data
916 mg/g
1.7 maximum

2430
No data
1213
1620
No data
No data
1216
1922

AM-3

Red Arrow Company

(Manitowoc, WI)
Red Arrow Company
Red Arrow Company
Red Arrow Company
Red Arrow Company
Red Arrow Company
Red Arrow Company
Red Arrow Company
Mastertaste, Inc.
(Monterey, TN)
Mastertaste, Inc.

4.254.85

1.82.1

0.30.8

1620

AM-3

Mastertaste, Inc.

4.3

2.2

Not detected

Not listed

List-A-Smoke

Mastertaste Inc.

2.02.5

7.08.0

1.754.25

58

Code 10-Poly

Mastertaste Inc.

2.3

10.3

3.22

Not listed

AM-10

Mastertaste Inc.

4.2

2.3

Not detected

Not listed

1291

Mastertaste Inc.

5.7

0.7

0.1

Not listed

Code V

Hickory Specialties
(Brentwood, TN)
Mastertaste Inc.
Mastertaste Inc.
Mastertaste Inc.
Mastertaste Inc.
Mastertaste Inc.
Mastertaste Inc.
Mastertaste Inc.
Mastertaste Inc.
Mastertaste Inc.
Mastertaste Inc.
Mastertaste Inc.
Mastertaste Inc.
Mastertaste Inc.

2.0

6.87.8

1.44.0

2.07.0

Paranjpye et al. (2004)

Faith et al. (1992)
Vitt et al. (2001)
Vitt et al. (2001)
Vitt et al. (2001)
Vitt et al. (2001)
Paranjpye et al. (2004)
Gedela, Escoubas, et al. (2007)
and Gedela, Gamble, et al. (2007)
Gedela, Escoubas, et al. (2007)
and Gedela, Gamble, et al. (2007)
Montazeri, Himelbloom, et al. (2013);
Montazeri, Oliveira, et al. (2013b)
Gedela, Escoubas, et al. (2007) and
Gedela, Gamble, et al. (2007)
Montazeri, Himelbloom, et al. (2013)
and Montazeri, Oliveira, et al. (2013)
Montazeri, Himelbloom, et al. (2013)
and Montazeri, Oliveira, et al. (2013)
Montazeri, Himelbloom, et al. (2013)
and Montazeri, Oliveira, et al. (2013)
EstradaMuoz et al. (1998)

23
6.17.0
23.0
4.15.0
23.0
23.0
5.16.0
6.17.0
6.17.0
23
33.2
4.14.3
67.4

4.55.9
01.4
6.07.4
3.04.4
6.07.4
6.07.4
1.52.9
01.4
01.4
4.55.9
3.43.7
1.51.8
01.4

05
05
05
20.125.0
05
05
05
05
05
0.30.6
0.30.6
0.30.6
0.30.6

151200.9
101150.9
101150.9
050.9
101150.9
51100.9
51100.9
101150.9
51100.9
151200
120132
110120
100110

Milly et
Milly et
Milly et
Milly et
Milly et
Milly et
Milly et
Milly et
Milly et
Milly et
Milly et
Milly et
Milly et

Charsol Supreme
Charsol Supreme
Charsol H-10
Charsol LFB Supreme Poly
Aro-Smoke P-50
CharOil
C-10
Zesti B

F1
F2
F3
F4
F5
F6
F7
F8
F9
S1
S2
S3
S4

Other studies suggest that the antimicrobial properties of liquid

smoke are not attributed to their phenol composition. Smoke fractions
from the Spanish food industries (L1, L2, L3 and S) were evaluated for
their antimicrobial properties at low temperature against A. hydrophila,
Yersinia enterocolitica and L. monocytogenes (Suen et al., 2001).
The pH of the liquid smokes was adjusted to neutrality and used at the
maximal concentration recommended by the manufacturer (0.4 to 4%)
and were subsequently incubated with 4 to 5 log10 CFU/mL of pathogen
for up to 21 days at 4 C in broth culture. All four extracts were effective
at eliminating or suppressing growth of A. hydrophila after 21 days.
Fractions L1 and S were bacteriostatic against Y. enterocolitica while fractions L2 and L3 were ineffective at reducing Y. enterocolitica numbers.
The most effective fraction against L. monocytogenes was fraction S
(3.2 log10 CFU/g reduction after 21 days) while fractions L1 and L2
exhibited slight inhibition by reducing cell counts by 2.1 log10 CFU/mL
and 1.9 log 10 CFU/mL, respectively. Fraction L3 did not inhibit
L. monocytogenes growth. The most active fraction, fraction S, was lowest in phenol concentration (23 mg/kg), but high in acid concentration
(23 mg/kg), while the least effective fraction L3 contained a high level
of phenols (99 mg/kg) suggesting that phenol concentration is not
indicative of the antimicrobial activity of liquid smoke.
3.3. Activity of carbonyls
Carbonyl compounds have also been suggested to contribute to the
antimicrobial properties of liquid smoke. Milly et al. (2005) determined
minimal inhibitory concentration (MIC) values of low phenolic liquid
smoke fractions against Listeria innocua M1 and several Gramnegative bacteria. The MIC values were determined for nine liquid

al. (2005)
al. (2005)
al. (2005)
al. (2005)
al. (2005)
al. (2005)
al. (2005)
al. (2005)
al. (2005)
al. (2008)
al. (2008)
al. (2008)
al. (2008)

smoke fractions (F19), eight of which had a phenol concentration of

0 to 5 mg/mL so that the antimicrobial effect of the non-phenolic compounds (i.e. carbonyl compounds and organic acids) could be evaluated.
The fraction F1 was found to be the most effective against the Gramnegative cocktail mixture with an MIC value of 1.5%. Fraction F1 had a
pH range of 2 to 3 and had the highest carbonyl content at 151 to
200.9 mg/mL. Fractions F3, F5 and F6 had MIC values of 2%, were similar
to F1 in pH and phenol level, but were lower in carbonyl content (101 to
150.9 mg/mL for F3 and F5; 51 to 100.9 mg/mL for F6). Fraction F4
which had a phenol content of 20.1 to 25.0 mg/mL, a pH of 4.1 to 5.0
and a carbonyl content of 0 to 50.9 mg/mL produced an MIC value of
3%. The least effective fractions, F7 (MIC 5%) and F9 (MIC 9%) contained
the same carbonyl level (51 to 100.9 mg/mL) but differed slightly in pH.
Fraction F7 with a lower pH range of 5.1 to 6.0 compared to that of F9,
pH 6.1 to 7.0, exhibited signicantly improved antimicrobial properties.
In assessing the antimicrobial properties of different liquid smoke
fractions, these data demonstrate the importance of knowing the concentrations of carbonyl compounds, acids and the pH values of these
products.
Three rened liquid smoke fractions along with a full strength fraction, Code 10-Poly (Kerry Ingredients and Flavors, Monterey, TN) were
tested in vitro for antilisterial properties. The three rened liquid
smoke fractions 1291, AM-10 and AM-3 possessed little to no phenols.
Using a disk diffusion assay it was determined that all fractions with
the exception of 1291 were effective against L. innocua. The largest inhibition zones were seen with Code 10-Poly which had a pH of 2.3, 10.3%
titratable acidity and total phenol content of 3.22 mg/mL. Fractions AM10 and AM-3 were also able to reduce the counts of L. innocua in culture
media tests, despite their having undetectable phenol levels suggesting

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201

Table 3
Efcacy of liquid smoke as an antimicrobial against select bacteria in vitro.
Fraction

Concentration

Bacteria

Method

Results

Reference

Aro Smoke P-50,

CharOil,
Charsol H-10,
Charsol LFB Supreme
Poly,
Charsol Supreme
Nine commercial LS from
Mastertaste F1F9

0100%

Listeria monocytogenes
ATCC 19115
Listeria innocua
ATCC 33090

MIC values determined by a

broth dilution method in BHI

Vitt et al. (2001)

100.5%

L. innocua M1

MICs determined by a broth

dilution method

100%
75%
50%
25%
Up to 2
manufacturers
recommended
concentration
0.4% L1
0.6% L2
4% L3
1% S
10.2%

L. innocua ATCC 33090

Disk diffusion

L. monocytogenes
CECT 932,
L. innocua CECT 4030

Agar dilution

L. monocytogenes
CECT 932

Flask containing LS were

inoculated and incubated at 4 C.

MIC values
Aro Smoke P-50 = 2.5%
CharOil = 5%
Charsol H-10 = 1.25%
Charsol LFB Supreme Poly = 1.25%
Charsol Supreme = 0.5%
MIC Values
F1 = 1.25%
F2 = 2%
F3 2%
F4 = 2%
F5 = 2%
F6 = 2%
F7 = 4%
F8 = 2%
F9 = 6%
Larger zones of inhibition were seen with
increasing LS concentration. Code 10-Poly
was the most effective fraction at all
conditions while 1291 was least effective.
Fractions L1, L4, S1 and S3 were not
effective. Fraction L2 was inhibitory at
0.6%, L3 at 8% for L. innocua and N8% for
L. monocytogenes, S2 at 1%
L1 and reduced Listeria by ~2 log CFU/mL
S reduced Listeria by 3.2 log CFU/mL and L3
was ineffective at reducing Listeria

Code 10-Poly
AM-3
AM-10
1291
Seven fractions from
Spanish food industry
L1, L2, L3, L4, S1, S2, and
S3 (all pH 7)
Four fractions from
Spanish food industry
L1, L2, L3, and S
Rice hull smoke extract

Four water based

commercial samples
and four concentrated
extracts from
commercial sources
Nine commercial LS from
Mastertaste F1F9

960.375%

100.5%

Lab generated from pyrolysis 1%0.05%

of beachwood chips

Four water based

960.375%
commercial samples and
four concentrated extracts
from commercial sources
Lab generated from pyrolysis 1%0.05%
of beachwood chips

Four water-based
commercial samples and
four concentrated extracts
from commercial sources

960.375%

Salmonella Typhimurium MICs were determined in

ATCC 14028
nutrient broth with phenol in a
microtiter plate
Salmonella Enteritidis
MICs were determined by a
PT 13A
microdilution method in
microtiter plates at 37 C.

MIC of rice hull smoke

condensate = 0.822%

Milly et al. (2005)

Montazeri, Himelbloom,
et al. (2013) and
Montazeri, Oliveira, et al.
(2013)
Suen (1998)

Suen et al. (2001)

Kim et al. (2012)

MIC values were 6% for 3 of the 4 water

Van Loo et al. (2012)
based samples and 12% for the fourth
sample. MICs were 0.5% for 3 of the 4
concentrated extracts; the nal concentrated
extract had an MIC value of 3%.
Gram negative cocktail of MIC values were determined
MIC Values
Milly et al. (2005)
Salmonella Muenster,
by a broth dilution method
F1 = 1.25%
Salmonella Seftenburg,
at 37 C.
F2 2%
Salmonella Typhimurium
F3 = 2%
and Escherichia coli 8677
F4 = 3%
F5 = 2%
F6 = 2%
F7 = 5%
F8 = 2%
F9 = 9%
E. coli ATCC 14948-K12
Smoke condensates were mixed Growth was delayed by 1 day at 0.125%, by Fretheim et al. (1980)
with nutrient broth agar followed 1.5 days at 0.166% and no growth was
by spread plating of E. coli. Plates observed at concentrations above 0.25%
were incubated at 30 C for
2 weeks.
E. coli O157:H7
MICs were determined by a
MIC were 6% for the water based samples Van Loo et al. (2012)
ATCC 43888
microdilution method in
and 0.5% for 3 of the 4 concentrated
microtiter plates
extracts, the nal concentrated extract had
an MIC value of 6%
Staphylococcus aureus
Smoke condensates were mixed No growth was observed at concentrations Fretheim et al. (1980)
ATCC 25923
with nutrient broth agar followed above 0.1%
by spread plating of E. coli. Plates
were incubated at 30 C for
2 weeks.
S. aureus ATCC 25923
MICs were determined by a
MIC were 6% for the water based samples Van Loo et al. (2012)
ATCC 6538
microdilution method in
and ~0.38% for concentrated extracts.
Mu50, MRSA
microtiter plates
Col, MRSA

that organic acids or carbonyls rather than phenols were involved in the
antilisterial properties of liquid smoke products they tested (Montazeri,
Himelbloom, Oliveira, Leigh, & Crapo, 2013).
Milly, Toledo, and Chen (2008) used high-end turkey rolls which
were whole parts of turkey breast formed to have no more than 40%
binders and broth added, low-end turkey rolls which were minced turkey breast parts with up to 60% binders and broth added before forming
and cooking, as well as roast beef cuts to evaluate low phenolic liquid

smoke fractions for antimicrobial properties. Turkey rolls and roast

beef cuts were rst inoculated with L. innocua M1, subsequently treated
with four different liquid smoke fractions (Mastertaste Inc., Brentwood,
TN), vacuum packed and refrigerated at 4 C for up to four weeks of storage. Samples were treated with 2 log10 CFU/25 cm2 of L. innocua M1 and
evaluated at 2 and 4 weeks of storage. Samples treated with smoke fractions S1, S2 and S3 had levels below the detection limit for L. innocua at
all the sampling times. Samples treated with S4 remained positive after

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J.M. Lingbeck et al. / Meat Science 97 (2014) 197206

Table 4
Efcacy of liquid smoke as an antimicrobial against several bacteria in food systems.
Liquid smoke fraction

Liquid smoke
concentration

Strain

Processing parameters

Result

Reference

CharSol Supreme

0.6% and 0.2%

Listeria monocytogenes
Scott A, V7, 101 M

60%

Listeria innocua

AM-3
AM-10

0.9%

L. innocua ATCC 33090

Listeria counts decreased after 3 days.

Estimated D-values are 4.5 h at 0.6% and
36 h for 0.2%
A 15 s dip resulted in a 3 log reduction
greater reductions were seen with
longer dip times
Both fractions reduced L. innocua to
b2 log CFU/g after 2 weeks.

Faith et al. (1992)

Charsol Supreme

Hot dog exudates containing LS were

inoculated with Listeria and incubated
at 25 C for up to 114 h.
100 g chum salmon samples were
brined, dipped for 5 min in LS,
inoculated and dried in a smoke house
Salmon strips were treated with LS,
inoculated, vacuum sealed, and stored
at 4 C for up to 49 days.

Fractions from
Spanish food
industries L1, L2,
L3 and S

100%, 1 min dip

L. monocytogenes CECT
932

Filets were brined, inoculated, treated

with LS and stored at 4 C for 21 days

CharSol C-10

100%
50%
25%
10%
dipped

L. monocytogenes strains
4121 and 1455

Salmon lets were brined, treated with

LS, inoculated and heat processed.

Zesti-B

100% dipped for 1

or 5 s or sprayed

L. monocytogenes Scott
A-2, V7-2, 39-2, 383-2

Zesti-B

100% sprayed

L. monocytogenes Scott
A-2, V7-2, 39-2, 383-2

Frankfurters formulated without lactate

and diacetate were dipped or sprayed in
LS, inoculated, vacuum packed and
stored at 1.7 C for 10 weeks.
Frankfurters formulated without lactate
and diacetate were sprayed in LS,
inoculated, vacuum packed and stored at
6.1 C for 10 weeks.

Fractions L1 and L2 immediately reduced

L. m to below detectable levels. Fraction
S slowly reduced Listeria and was below
detectable levels by 21 days. Fraction L3
did not show any inhibitory affects.
Internal minimum lethality temperatures
were N82.8 C in untreated salmon steaks,
67.2 C generated smoke was applied
throughout the entire smoking process
or N 80 C when smoke was only applied
during the last half of the process, 58.9 C
when dipped in 100% CharSol C-10 and
62.8 C, 68.9 C and 72.8 C with 50%, 25%
and 10% LS.
L. m. was reduced to undetectable
numbers after 4 weeks with all
treatments

Zesti-B AM-3

100%, 1 s dip

L. monocytogenes Scott
A-2, V7-2, 39-2, 383-2

AM-3

100% sprayed to
equal 1.8 mL per
frank

CharSol-10

100%, dipped

Zesti Smoke

Formulated into
franks at 10, 5,
and 2.5% (wt/wt)

L. monocytogenes ARS
V67, ARS V72, ARS V113,
ARS V125, ARS V105,
LCDC 81861
L. monocytogenes LCDC
81861, M1, M2, M5, C6,
serotype 4b derived
ATCC 19115
L. monocytogenes

Zesti-B

100%, 1 s dip

L. monocytogenes Scott
A-2, V7-2, 39-2, 383-2

Four commercial
LS from Mastertaste
S1, S2, S3 and S4

100%, 60s dip

L. innocua M1

Four commercial
LS from Mastertaste
S1, S2, S3 and S4

100%, 60s dip

L. innocua M1

Frankfurters were formulated without

lactate or diacetate, dipped in LS,
inoculated, vacuum sealed, heat
pasteurized for 1 min at 37 C, chilled
and stored at 6.1 C for 10 weeks
Frankfurters were formulated without
lactate or diacetate, sprayed with LS,
inoculated, sealed and stored at 4 C for
140 days.
Frankfurters were inoculated, dipped in
LS, vacuum packed and stored at 4 C for
72 h.
Frankfurters were inoculated, vacuum
packed and stored at 4 C for 12 weeks.

Deli turkey formulated without lactate or

diacetate were dipped in LS, inoculated,
vacuum packed, heat pasteurized 60s at
93.3 C, chilled and stored at 6.1 C for
10 weeks
High end turkey rolls (containing
less-than 40% binders and broth added),
low end turkey rolls (up to 60% binders
and broth added) were dipped in LS,
inoculated with L. i. in a marked 25 cm2
area, vacuum sealed and stored at 4 C
for 4 weeks.
Roast beef cuts were dipped in LS, inoculated with L. i. in a marked 25 cm2 area,
vacuum sealed and stored at 4 C for
4 weeks.

Vitt et al. (2001)

Montazeri,
Himelbloom, et al.
(2013) and
Montazeri, Oliveira,
et al. (2013)
Suen et al. (2003)

Poysky et al. (1997)

Gedela, Escoubas
et al. (2007)

L. m. was reduced to undetectable levels

after 1 week at log 1 inoculum levels, at
log 2 the levels declined slowly and were
undetectable after 4 weeks. The log 3
inoculum slowly increased over 10 weeks
to 0.8 logs higher that inoculum while
control increased by 7 logs
L. m. levels were undetectable after
3 weeks with both Zesti-B and AM-3
fractions

Gedela, Escoubas
et al. (2007)

L. m. was reduced by nearly 3 logs after

30 days and continued to slowly decline
for up to 130 days

Martin et al. (2010)

L. m. was reduced initially by at least

1 log and was undetectable after 72 h.

Messina et al. (1988)

Frankfurters formulated with 2.5% LS

saw a 0.5 log CFU/mL reduction when
inoculated at high L. m. levels and ~2 log
CFU/mL when inoculated with lower
levels of Listeria. Frankfurters formulated
with 5% LS saw greater reductions and
were listericidal after 6 weeks. 10% LS
was listericidal after 4 weeks.
L. m. levels were reduced by 2 logs after
2 weeks and remained low after
10 weeks

Morey et al. (2012)

L. i. levels were reduced to undetectable

levels after 2 and 4 weeks of storage on
both high and low end turkey rolls with
all LS samples tested, with the exception
of S4 on low end rolls. Only a small
reduction was seen after two weeks
(0.53 log CFU) but was not detected after
4 weeks.
L. i. levels were reduced to undetectable
levels after 2 and 4 weeks of storage with
all fractions tested.

Milly et al. (2008)

Gedela, Gamble,
et al. (2007)

Gedela, Gamble,
et al. (2007)

Milly et al. (2008)

J.M. Lingbeck et al. / Meat Science 97 (2014) 197206

203

Table 4 (continued)
Liquid smoke fraction

Liquid smoke
concentration

Strain

Processing parameters

Result

Reference

Code V

8%

Escherichia coli O157:H7

Growth was reduced by 2.3 log CFU/g

Estrada-Muoz et al.
(1998)

C10

Staphylococcus aureus

Charsol Supreme

75%

No reduction after 5 days

None detected after 5 days
None detected after 3 days
None detected after 3 days
1.7 log reduction after 3 h compared to
control
No reduction after 15 h compared to
control, however no enterotoxins were
present in LS treated samples
0.74 log reduction and no enterotoxins
were detected

Paranjpye et al.
(2004)

Charsol Supreme

25%
50%
75%
100%
1.25%

Beef trimmings were inoculated and

treated with LS after which they were
ground and formed into 7090 g patties.
Patties were packaged, and stored at 4 C
for 3 days.
Brined strips were dipped for 1 min,
inoculated, and processed at 30 C for
35 days

Cocktail mixture of
S. aureus ATCC 27664
(enterotoxin E), ATCC
13565 (enterotoxin A),
and ATCC 12660
Cocktail mixture of
S. aureus ATCC 27664
(enterotoxin E), ATCC
13565 (enterotoxin A),
and ATCC 12660

Inoculated, ground pork bellies were

treated with LS and heated to 50 C over
6 h followed by cooling to 7.2 C for
315 h
50 g pieces were inoculated and heated
to 50 C over 6 h. LS was sprayed at
hours 4 or 5. Sample were cooled to
7.2 C for 15 h.

both two and four weeks of cold storage. All liquid smoke fractions tested contained similar phenol concentrations (0.3 to 0.6 mg/mL) while
carbonyl concentrations ranged from 110 to 200 mg/mL for fractions
S1, S2 and S3 and from 100 to 110 mg/mL for S4. Fraction S4 was also
lower in acidity (0 to 1.4%) and higher in pH (6 to 7.4) than the other
samples (acidity range1.5 to 5.9% and pH 2 to 4.3). Again, the data
suggest that phenols contribute little to the bacteriostatic properties of
liquid smoke and that carbonyl compounds and acidity levels are
important in assessing the antimicrobial properties of liquid smoke
products. The authors also reported that food product composition
also plays an important role in bacterial survival in that the lower end
turkey rolls were apparently able to support bacterial growth better
than the high end turkey rolls or roast beef cuts (Milly, Toledo, &
Chen, 2008).
Liquid smoke has demonstrated ability at reducing foodborne
pathogens including L. monocytogenes, E. coli, S. aureus and Salmonella
in RTE foods and other food products. The exact mechanism of action
is unknown, and both phenolic compounds and carbonyl compounds
are thought to contribute to its antimicrobial properties. Table 4 lists
the chemical components of several commercial liquid smokes and
other liquid smoke fractions discussed below.
4. Antimicrobial activity of liquid smoke against Listeria
L. monocytogenes is a Gram-positive food borne bacterium. Listeria
has the ability to grow to infective doses at low temperatures, high
salt concentrations, and under acidic and microaerophilic conditions,
which make it difcult to control on many ready-to-eat (RTE), refrigerated foods. The infective dose of L. monocytogenes is dependent upon
several factors including the bacterial strain and susceptibility of the individual to the bacterium, but it can be as low as 1000 bacteria (SchmidHempel & Frank, 2007). Consumption of an infective dose of Listeria
may lead to the food borne infection, listeriosis, in humans especially
in older adults or persons who are immunocompromised as well as
pregnant women and newborns. Symptoms of listeriosis include fever,
muscle aches, nausea or diarrhea and may result in meningitis, premature labor, miscarriage or even death. In the US, 1651 cases of listeriosis
and 292 deaths or fetal losses as a result of Listeria infection were reported between 2009 and 2011 (CDC, 2011).
4.1. In vitro effects on Listeria
Pittman et al. (2012) studied the effects of individual stressors of
cold smoking including freezing, thawing, salt exposure, exposure to
liquid smoke and cold storage on virulent and avirulent Listeria. Two

Taormina and
Bartholomew (2005)

Taormina and
Bartholomew (2005)

strains, virulent HCC7 and avirulent HCC23, both in mid-log growth

were examined by transmission electron microscopy (TEM) to identify
changes in cell wall integrity due to exposure to a simulated cold
smoking process in brain heart infusion (BHI) culture medium. Bacteria
were subjected to a sequence of stresses: a freeze thaw cycle (20 C
for 2 h followed by room temperature thawing for 1 h); exposure to
6% NaCl at 30 C for 1 h, exposure to 0.6% liquid smoke at 30 C for 1 h
and nally anaerobic storage at 2 C for 16 h. The authors found that
the avirulent strain was more susceptible than the virulent strain to all
of the above conditions of the cold-smoking process; the virulent strain
was only signicantly affected by application of liquid smoke and anaerobic storage. The results suggested that these two different strains
utilize different mechanisms to adapt to the above stresses. More importantly, they found that both strains can survive the effects of this
simulated cold smoking process, emphasizing the need for additional
effective but all-natural antimicrobial interventions to control this pathogen (Pittman et al., 2012).
4.2. Antilisterial effects in ready-to-eat meats
Ready-to-eat (RTE) meats such as luncheon meat and frankfurters treated with liquid smoke have shown reduced levels of
L. monocytogenes. Beef franks were inoculated by immersion into a
3 log10 CFU/mL cocktail of six strains of L. monocytogenes. After air
drying, frankfurters were dipped in full strength CharSol-10, air dried,
vacuum packed and stored at 4 C. After 72 h, frankfurters treated
with CharSol-10 exhibited a 3 log10 reduction in L. monocytogenes
counts compared to control samples whose bacterial count remained
unchanged (Messina, Ahmad, Marchello, Gerba, & Paquette, 1988).
Spray application of liquid smoke was also successful in reducing
L. monocytogenes levels on frankfurters formulated without the typical
Listeria growth inhibitors of sodium lactate and sodium diacetate.
Frankfurters were sprayed with the commercial liquid smoke fraction AM-3 followed by inoculation with a six strain cocktail of
L. monocytogenes at a concentration of 5 log10 CFU/mL. Frankfurters
were stored at 4 C and bacterial levels were monitored at 2, 24 and
48 h. Liquid smoke treated samples showed a continuous decline of
L. monocytogenes with time until no detectable levels were seen at 48
h (Martin et al., 2010). An extended shelf life study in which lactate/
diacetate free frankfurters were sprayed with AM-3 and inoculated
with a cocktail of L. monocytogenes, showed that liquid smoke could
suppress L. monocytogenes on frankfurters vacuum packed and stored
at 4 C for up to 130 days (Martin et al., 2010). In a similar study
lactate/diacetate free frankfurters were inoculated with 3 log10 CFU/mL
of a four strain cocktail of L. monocytogenes, sprayed with the liquid

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J.M. Lingbeck et al. / Meat Science 97 (2014) 197206

smoke fraction Zesti B (Mastertaste, Monterey, TN) and stored at

the abuse temperature of 6.1 C for up to 10 weeks. Levels of
L. monocytogenes decreased after one week of storage, but thereafter increased by only 0.8 log10 CFU/g above the initial inoculum level by the
end of the 10 weeks as compared to a 7 log10 increase in untreated
franks (Gedela, Escoubas, & Muriana, 2007). The authors also tested
the effectiveness of a dip application of liquid smoke to frankfurters.
Retail franks (containing lactate and diacetate) which had previously
been shown to permit growth of L. monocytogenes at 1.7 C after
6 weeks were dipped in Zesti-B liquid smoke for 1 to 90 s followed by
inoculation with 1 log10 CFU of a Listeria cocktail. Viable counts were
reduced to below the detection level after 4 weeks of storage for all
dip times tested whereas untreated controls showed a greater than
4 log10 increase (Gedela, Escoubas, et al., 2007; Gedela, Gamble, et al.,
2007).
A combination of post-process heat pasteurization and a liquid
smoke containing reduced acid levels and low phenolic concentrations
was able to reduce L. monocytogenes on frankfurters and deli turkey. Of
two smoke extracts tested, the product coded as Zesti-B had 3.5 to 5.6%
acidity, a pH 2.5 to 3.3, with a maximum of 1.7 mg/mL of phenols and a
carbonyl concentration of 19 to 22 g/100 mL while the product coded as
AM-3, a fraction lighter in color and reduced smoke avor, contained
1.8 to 2.1% acidity, a pH 4.25 to 4.85, with a 0.3 to 0.8 mg/mL phenol
concentration and a carbonyl concentration of 16 to 20 g/100 mL.
Frankfurters were formulated without lactate/diacetate and dipped for
1 s into Zesti-B or AM-3, inoculated with a four strain mixture of
L. monocytogenes at 5 log10 CFU/sample, vacuum packed, pasteurized
at 73.9 C for 1 min and stored at the mild abuse temperature of 6.1 C
for up to 10 weeks. No Listeria was detected immediately after processing and L. monocytogenes levels remained below detectable limits during 10 weeks of storage. Turkey breast chubs formulated without
lactate/diacetate or nitrate underwent similar treatment. Samples
treated with liquid smoke 60 to 30 s post-process pasteurization resulted in 2 to 3 log 10 reductions of Listeria by week two.
L. monocytogenes levels remained consistently low for the remainder of
the experiment (Gedela, Gamble, Macwana, Escoubas, & Mariana, 2007).
The use of liquid smoke as an ingredient, as opposed to topical application, has also been shown to be effective in reducing L. monocytogenes.
Frankfurters, formulated with 2.5%, 5% or 10% (wt/wt) of the liquid
smoke fraction Zesti Smoke (Kerry Ingredients and Flavors, TN), were
inoculated with either 4 or 8 log10 CFU/mL of L. monocytogenes, vacuum
packed and stored at 4 C for 12 weeks. At select sampling times the
frankfurters were rinsed and the rinsates were plated on Modied
Oxford agar to detect the presence of Listeria. The liquid smoke incorporated as an ingredient in frankfurters was able to suppress the growth of
L. monocytogenes inoculated at both high (8 log10 CFU/mL) and low inoculation levels (4 log10 CFU/mL) at its lowest concentration (2.5%)
compared to untreated samples. Inoculated frankfurters not formulated
with liquid smoke permitted growth of L. monocytogenes up to more
than 8 log10 CFU/mL in 12 weeks for both high and low inoculation
levels. The addition of 2.5% liquid smoke to the frankfurters reduced
L. monocytogenes levels compared to control levels to 6.2 log10 CFU/mL
for the low inoculation level and 7.47 log10 CFU/mL for the high level
inoculation. Further suppression was seen with increasing liquid
smoke concentrations. However, frankfurters containing 10% liquid
smoke were somewhat less acceptable in sensory panel tests as compared to frankfurters containing 2.5% or 5% liquid smoke (Morey,
Bratcher, Singh, & McKee, 2012).
4.3. Genetic basis of the antimicrobial effects of liquid smoke on Listeria
Liquid smoke has a demonstrated success at limiting the growth of
Listeria. Using proteomics Guilbaud et al. (2008) examined the effects
of liquid smoke on L. monocytogenes. Listeria was exposed to 30 g/mL
of phenol for 2 h which resulted in the upregulation of ClpP-2, a subunit
of the general stress protein ClpP which is required for virulence

expression in L. monocytogenes. Also affected were proteins involved

in metabolic pathways including Lmo355 and Lmo2829, proteins involved in membrane bioengineering and lipid metabolism, suggesting
that liquid smoke affects the synthesis of the cell membrane. In addition
liquid smoke reduced the hemolytic activity of Listeria and may reduce
the virulence of Listeria. This may partially explain the extremely limited
number of food borne outbreaks involving cold-smoked sh.
5. Effects of liquid smoke on Salmonella spp.
Salmonella is a Gram-negative food borne bacterium. Salmonella contamination is especially prevalent in raw poultry and eggs, but the organism can contaminate a wide variety of different foods including dairy,
meat and raw vegetables and fruits and animal feeds (Maciorowski,
Jones, Pillai, & Ricke, 2004; Foley et al., 2011; Howard, O'Bryan,
Crandall, & Ricke, 2012; Finstad, O'Bryan, Marcy, Crandall, & Ricke,
2012). The infectious dose of Salmonella is approximately 5 log10
organisms (Schmid-Hempel & Frank, 2007). Symptoms of Salmonella
infection include fever, cramps, vomiting and diarrhea. Generally symptoms only last a few days and leave no lasting effects; however, in some
cases life threatening complications may arise (Ricke, Koo, Foley, &
Nayak, 2013).
A liquid smoke generated from the pyrolysis of rice hulls was shown
to be bactericidal against Salmonella Typhimurium in a disk diffusion
assay at concentrations ranging from 0.1% to 1%. Larger zones of inhibition were seen at higher rice hull liquid smoke concentrations. The MIC
was 0.822% (v/v) as determined by a broth dilution assay. Interestingly,
rice hull liquid smoke also enhanced survival of mice infected with a
lethal dose of Salmonella. The Balb/c mice were fed a commercial
chow diet containing 1% rice hull liquid smoke for 14 days after which
they were injected with 5 log10 CFU of Salmonella Typhimurium. Mice
fed rice hull liquid smoke survived twice as long (about14 days)
compared to mice that were fed the untreated diet who survived approximately 7 days (Kim et al., 2012).
Van Loo et al. (2012) determined the MICs of seven commercial
liquid smoke samples from three commercial manufacturers against S.
Typhimurium using a microdilution method. The liquid smoke samples
were separated into two groups. Group I contained water based liquid
smokes extracted from woods of hickory, mesquite, apple and pecan.
Group II were concentrated liquid smokes extracted from hickory and
mesquite. Group II liquid smokes were more effective at reducing
Salmonella with MIC values ranging from 0.5% to 4%. Group I was less
effective and produced MIC values from 6% to 12%. Similar results
were seen with nine commercial liquid smoke samples (Mastertaste,
Brentwood, TN) on a cocktail of Gram-negative bacteria consisting of
S. Muenster, S. Seftenburg, S. Typhimurium and E. coli. MIC values
ranged from 1.5% to 9% (Milly et al., 2005).
6. Effects of liquid smoke on E. coli
E. coli is a Gram-negative bacterium found in the intestines of most
animals including humans. Although most strains of E. coli are innocuous, some strains such as the Shiga toxin producing strain O157:H7
(STEC) and non O157:H7 Shiga toxin producing strains can cause illness
if ingested (Jaeger, 1999). E. coli O157:H7 is infectious at concentrations
as low as 10 organisms (Schmid-Hempel & Frank, 2007). Complications
arising from STEC strains of E. coli infections include cramps, vomiting,
bloody diarrhea, and blood in the urine, with severe infections leading
to kidney failure (Belongia et al., 1991). Contamination with STEC strains
of E. coli is often found in undercooked meat especially beef, unpasteurized dairy products and raw fruits and vegetables.
6.1. In vitro effects of liquid smoke on E. coli
The antimicrobial properties of smoke condensates in concentrations ranging from 0.05% to 1% were evaluated against STEC strains of

J.M. Lingbeck et al. / Meat Science 97 (2014) 197206

E. coli. These strains were able to grow in up to 0.1% smoke while at

0.125%, growth was delayed by 1 day and at 0.25% no growth was
observed (Fretheim, Granum, & Vold, 1980). Values for MICs for several
commercial liquid smokes against E. coli O157:H7 were reported by Van
Loo et al. (2012). Effective concentrations ranged from 6% to as low as
0.75% depending upon the commercial brand of liquid smoke.

205

avor and also has inhibitory effects on food borne pathogens. The
preservative effect of liquid smokes is achieved by antimicrobial and
antioxidant compounds such as aldehydes, carboxylic acids and phenols.
Liquid smoke also has several advantages over traditional smoking techniques including ease of application, speed of smoking process, good reproducibility of desired characteristics obtained in the nal smoked
food, and omission of hazardous polycyclic aromatic hydrocarbons.

6.2. Effects of liquid smoke on E. coli in beef

The effects of liquid smoke on E. coli in a model meat system have
been reported. Beef trimmings were inoculated with 7 log10 CFU/g of
the STEC strain E. coli O157:H7 and treated with a nal concentration
of 8% of the liquid smoke fraction Code V (Hickory Specialties,
Brentwood, TN). Trimmings were then ground, formed into patties,
heat sealed and stored at 4 C. Liquid smoke treated trimmings showed
a 2.3 log10 CFU/g reduction after three days of refrigerated storage compared to untreated samples (Estrada-Muoz, Boyle, & Marsden, 1998).
7. Effect of liquid smoke on Staphylococcus
Staphylococcus is a Gram-positive spherical-shaped bacterium
appearing in grape-like clusters (Betts, 2010). S. aureus is salt tolerant
and produces heat-stable enterotoxins which are responsible for
staphylococcal foodborne illnesses. Foodborne illness due to S. aureus
is a result of eating foods contaminated with toxins produced by the
bacteria as opposed to consumption of the bacteria itself (Betts,
2010); it is generally agreed that populations of S. aureus need to
reach 5 to 6 log10 organisms for toxin production to occur (SchmidHempel & Frank, 2007). Symptoms of S. aureus infection include nausea,
vomiting and diarrhea which generally last 1 to 3 days. Foods commonly contaminated with S. aureus are prepared foods that require no additional cooking such as salads of tuna, egg, chicken, potato and macaroni,
sandwiches, bakery products containing a cream lling as well as meat,
poultry and eggs (Betts, 2010).
Bacon has been shown to also harbor staphylococci (Taormina &
Bartholomew, 2005). In order to validate that bacon processing does
not permit the growth of S. aureus, cured, raw pork bellies were ground
and inoculated with a cocktail of S. aureus followed by application of
Charsol Supreme to a nal concentration of 1.25%. Samples were slowly
heated over the course of 6 h to 50 C, the peak smoking temperature
for bacon, and cooled to 7.2 C in 3 h to simulate smoking and blast chilling that occurs in commercial plants. S. aureus populations increased by
only 0.68 log10 CFU/g in liquid smoke treated samples, when compared
to control samples which had an increase of 2.38 log10 CFU/g. When
cooling times were increased from 6 to 15 h S. aureus grew by 4 log10
CFU/g in both smoke treated and untreated samples, however no
enterotoxins were detected in samples treated with liquid smoke. In a
separate experiment, whole pork bellies were sliced into 50 g pieces
and inoculated by injection of the bacterial cocktail to a nal inoculation
concentration of approximately 2 log10 CFU/g. The pieces were then
subjected to simulated smoking. A 75% solution of Charsol Supreme
was sprayed after 4 and 5 h of the smoking process after which the
samples were cooled to 7.2 C over the course of 15 h and bacterial populations were enumerated. After simulated smoking and extended
cooling S. aureus populations were reduced to 0.74 0.53 log10 CFU/g
and samples were negative for staphylococcal enterotoxins (Taormina
& Bartholomew, 2005).
8. Conclusions
Liquid smoke is an effective antimicrobial against an array of bacterial pathogens as demonstrated in both broth culture and food systems.
Commercial use of liquid smoke in the food industry may satisfy consumer demand for all-natural foods while still maintaining their safety.
Liquid smoke is being used more frequently in preserving protein-based
foods, namely meat, sh, and cheese, because it imparts a pleasant

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