Sie sind auf Seite 1von 8

Perspectives on the Use of Organic Acids

and Short Chain Fatty Acids as Antimicrobials

S. C. Ricke1

Department of Poultry Science, Texas A&M University, College Station, Texas 77843-2472

ABSTRACT Organic acids have a long history of being Although this situation has implications regarding the
utilized as food additives and preservatives for pre- use of organic acids, it may only apply to circumstances
venting food deterioration and extending the shelf life of in which reduced acid levels have induced resistance and
perishable food ingredients. Specific organic acids have virulence mechanisms in exposed organisms. Evaluating
also been used to control microbial contamination and effectiveness of organic acids for specific applications re-
dissemination of foodborne pathogens in preharvest and quires more understanding general and specific stress
postharvest food production and processing. The antibac-
response capabilities of foodborne pathogens. Develop-
terial mechanism(s) for organic acids are not fully under-

Downloaded from http://ps.oxfordjournals.org/ by bahram afsharmanesh on June 25, 2014


ment and application of molecular tools to study patho-
stood, and activity may vary depending on physiological
status of the organism and the physicochemical character- gen behavior in preharvest and postharvest food produc-
istics of the external environment. An emerging potential tion environments will enable dissection of specific bacte-
problem is that organic acids have been observed to en- rial genetic regulation involved in response to organic
hance survivability of acid sensitive pathogens exposed acids. This could lead to the development of more tar-
to low pH by induction of an acid tolerance response and geted strategies to control foodborne pathogens with or-
that acid tolerance may be linked to increased virulence. ganic acids.
(Key words: organic acid, antimicrobial, short-chain fatty acid, stress)
2003 Poultry Science 82:632–639

INTRODUCTION given the equally intensive public concern regarding


prevalence of foodborne pathogens in food animals dur-
Antibiotic supplementation in farm animal diets to im- ing preharvest and postharvest phases of animal produc-
prove animal growth performance and efficiency of feed tion, limiting pathogen dissemination with antimicrobials
conversion, historically, has been theorized as a means remains a desired goal. This has lead to somewhat of
of establishing food animals as reservoirs for antibiotic- a renaissance in research activity on the discovery and
resistant pathogens isolated from humans (Smith, 1968; application of nonantibiotic chemical compounds capable
Levy et al., 1976; Armstrong, 1984; Neu, 1992). Given of either killing microorganisms outright or at the very
the rising concerns associated with increased antibiotic least retarding growth sufficiently to limit their dissemi-
resistance in bacterial pathogens making treatment of nation.
clinical diseases more difficult, antibiotic use in animal
management in the U.S. and Europe has become more of ORGANIC ACIDS
a controversial issue (Glynn et al., 1998; Monnet, 1999;
Koutsolioutsou et al., 2001). This has resulted in a height- Classification and Uses
ened awareness regarding exposure of foodborne patho-
gens to therapeutic levels of antibiotics during preharvest Organic acids have a long history of being utilized
phases of food animal production (Aarestrup and Weg- as food additives and preservatives for preventing food
ener, 1999; Threlfall et al., 2000). Such concern necessitates deterioration and extending the shelf life of perishable
a certain perceived inevitability for potential decreases food ingredients. As a group these compounds primarily
in the uses of traditional antibiotics for routine nonclinical include the saturated straight-chain monocarboxylic acids
applications in animal management practices. However, and their respective derivatives (unsaturated, hydroxylic,
phenolic, and multicarboxylic versions) and are often ge-
nerically referred to as fatty acids, volatile fatty acids, or
2003 Poultry Science Association, Inc.
Received for publication October 9, 2002.
Accepted for publication December 5, 2002.
1
To whom correspondence should be addressed: sricke@poultry. Abbreviation Key: ATP = adenosine triphosphate; SCFA = short-
tamu.edu. chain fatty acids.

632
SYMPOSIUM: USE OF ANTIMICROBIALS IN PRODUCTION 633
weak or carboxylic acids Cherrington et al., 1991). Several acids serve as uncouplers that generally dissipate pH
of these organic acid compounds are used as direct addi- and electrical gradients across cell membranes (Sheu and
tives incorporated into human foods or can accumulate Freese, 1972; Sheu et al., 1972, 1975; Freese et al., 1973;
over time as a consequence of the fermentation activity Salmond et al., 1984; Russell, 1992; Axe and Bailey, 1995;
of indigenous or starter cultures added to certain dairy, Davidson, 2001).
vegetable, and meat products. In addition, acid sprays Russell (1992) has hypothesized that anion accumula-
have been incorporated as sanitizers during meat pro- tion is the primary toxic effect of organic acids and that
cessing (Acuff et al., 1987; Cherrington et al., 1991; Dick- some organisms are more resistant to organic acids be-
son, 1992; Hardin et al., 1995; Dorsa, 1997). Some organic cause they are capable of allowing their internal pH to
acids, particularly the short-chain fatty acids (SCFA), ace- decline. Less direct antibacterial activities have also been
tate, propionate, and butyrate, are produced in millimolar attributed to organic acids and include interference with
quantities in the gastrointestinal tracts of food animals nutrient transport, cytoplasmic membrane damage re-
and humans and characteristically occur in high concen- sulting in leakage, disruption of outer membrane perme-
trations in regions where strictly anaerobic microflora are ability, and influencing macromolecular synthesis (Cher-
predominant. In the food animal industry, organic acids rington et al., 1991; Denyer and Stewart, 1998; Alakomi
were originally added to animal feeds to serve as fungi- et al., 2000; Davidson, 2001).
stats (Paster, 1979; Dixon and Hamilton, 1981), but in The mechanisms associated with these activities have

Downloaded from http://ps.oxfordjournals.org/ by bahram afsharmanesh on June 25, 2014


the past 30 yr, formic and propionic acids and various been difficult to establish due to the complex nature of
combinations have also been examined for potential bac- the interaction between energy dissipation and disruption
tericidal activity in feeds and feed ingredients contami- of ATP generation capabilities of the bacteria. Conse-
nated with foodborne pathogens, particularly Salmonella quently, bactericidal concentrations of organic acids may
spp. (Khan and Katamay, 1969; Vanderwal, 1979; Wil- be due to the combination of dissipation of proton-motive
liams, 1981; Hinton and Linton, 1988; Humphrey and force and inability to maintain internal pH followed by
Lanning, 1988; Izat et al., 1990; McHan and Shotts, 1992; denaturation of acid-sensitive proteins and DNA. Suble-
Berchieri and Barrow, 1996; Thompson and Hinton, 1997). thal concentrations may elicit their effects on overall cell
physiology and lead to responses such as enlargement of
Antibacterial Activities of Organic Acids bacterial cell size (Thompson and Hinton, 1996).

In general, potential bacterial targets of biocidal com- Factors that Influence Toxicity
pounds include the cell wall, cytoplasmic membrane, and Efficacy of Organic Acids
specific metabolic functions in the cytoplasm associated
with replication, protein synthesis, and function (Denyer The significant affects and the toxicity of organic acids
and Stewart, 1998; Davidson, 2001). Although the antibac- (either directly or indirectly) for foodborne pathogens
terial mechanism(s) for organic acids are not fully under- have not been clearly elucidated. This is in part due to
stood, they are capable of exhibiting bacteriostatic and complexities involved in designing and conducting the
bactericidal properties depending on the physiological experiments that precisely delineate all of the contribut-
status of the organism and the physicochemical character- ing factors. Direct assessment and comparison of antibac-
istics of the external environment. Given the weak acid terial potency of individual acids can be elusive because
nature of most of these compounds, pH is considered a of the influence of the physical chemistry of respective
primary determinant of effectiveness because it affects the acids, the bacterial species in question, growth conditions
concentration of undissociated acid formed (Davidson, or media composition, and the growth phase (Cherring-
2001). It has been traditionally assumed that undissoci- ton et al., 1991; Lasko et al., 2000). In addition, specific
ated forms of organic acids can easily penetrate the lipid quantitative in vitro responses do not necessarily translate
membrane of the bacterial cell and once internalized into to all in vivo possibilities for a particular organism. This
the neutral pH of the cell cytoplasm dissociate into anions is partially due to the fact that many of the common
and protons (Eklund, 1983, 1985; Salmond et al., 1984; foodborne pathogens such as Salmonella spp. can grow
Cherrington et al., 1990, 1991; Davidson, 2001). Genera- in a multitude of ecosystems and, thus, in vivo growth
tion of both of these species potentially presents problems conditions when exposure to organic acids occurs can
for bacteria that must maintain a near neutral pH cyto- vary widely. It is entirely possible that during a life cycle,
plasm to sustain functional macromolecules. Export of Salmonella can grow aerobically and within a short time
excess protons requires consumption of cellular adeno- frame be forced to switch to an anaerobic metabolism to
sine triphosphate (ATP) and may result in depletion of survive and potentially colonize a highly fermentative
cellular energy (Davidson, 2001). environment such as the gastrointestinal tract. Under
Other toxicity mechanisms have been proposed that these conditions not only would Salmonella be required to
attribute membrane uncoupling capabilities for organic survive high concentrations of SCFA produced by other
acids. It has been speculated that organic acids interfere organisms, but it would also be generating and exporting
with cytoplasmic membrane structure and membrane fermentation organic acids of its own. Distinguishing spe-
proteins such that electron transport is uncoupled and cific organic acid toxic mechanisms in foodborne patho-
subsequent ATP production is reduced or that organic gens will continue to be difficult to achieve until genetic
634 RICKE

techniques can be applied to screen for differential physi- acids simply disappear in the gastrointestinal tract before
ological responses in selected mutants exposed to the reaching locations inhabited by pathogens. Using radio-
multitude of potential environmental backgrounds. actively labeled propionate, Hume et al. (1993b) demon-
Efficacy of organic acids can also be highly influenced strated that most of the propionic acid originating from
by extrinsic factors associated with the environment that the treated feed is metabolized and absorbed in the fore-
they are being added to. An example of this complexity gut of the chicken (crop, gizzard, and proventriculus) and
is the use of propionic acid and its salt forms in animal does not reach the small and intestine or the cecum in
and poultry feeds as an anti-Salmonella and antifungal sufficient quantities to be effective.
compound. It has been suggested that the presence of
propionate in feeds limits Salmonella and fungal contami- Bacterial Mechanisms of Resistance to
nation in the ingredients added to feed and mixed feeds Antibacterial Activities of Organic Acids
during storage. However, several factors can alter the
effective concentration in the feed. Inconsistent antimicro- In addition to the environmental factors that can influ-
bial effects of propionic acid on fungal populations in ence the efficacy of organic acids as antimicrobial chemi-
feed has been attributed to possible buffering and conver- cals, inherent resistance of the target microorganism to
sion to its less active form by protein ingredients such as these compounds is also a factor (Davidson, 2001). Micro-
soybean meal as well as batch differences in primary feed bial sensitivity to antimicrobial agents determines relative

Downloaded from http://ps.oxfordjournals.org/ by bahram afsharmanesh on June 25, 2014


components such as corn meal (Dixon and Hamilton, tolerance and is dependent on the type of organism (fungi,
1981; Tabib et al., 1984). Feed composition may also influ- virus, or bacteria), strain-to-strain variation and form (en-
ence sensitivity of the organism. For example, there ap- dospore versus vegetative cell) with vegetative cells being
pears to be an interaction between water activity and more susceptible and bacterial spores being the most re-
survival of salmonellae with more sustained survival oc- sistant (Heinzel, 1998). Metabolically, microbial resistance
curring in populations adapted to lower water activities may be transient in nature or expressed in a more stable
(Mossel and Koopman, 1965; Liu et al., 1969; Carlson and form, and mechanisms of resistance can include exclusion
Snoeyenbos, 1970; Juven et al., 1984; Rouse et al., 1988). of the antimicrobial chemical, secretion of the toxic com-
This becomes a factor in evaluating organic acid efficacy pound, or metabolic detoxification (Heinzel, 1998).
because it has been demonstrated that after 8 wk only Genetic encoding in bacteria for resistance to antimicro-
minimal reductions in salmonellae populations occur and bial compounds can be fairly mobile within a population
viable populations can be recovered from feed 3 to 18 mo of the same organism and can potentially be transferred
post inoculation (Williams and Benson, 1978; Davies and among populations of genetically different organisms.
Wray, 1996; Ha et al., 1998a,b). The resulting physiological Although less is known about organic acids, genetic
status of the pathogen contained in the feed over long mechanisms in microorganisms for resistance to antibiot-
periods may also influence organic acid efficacy as bacte- ics and inorganic chemical-based disinfectants have been
rial cells in stationary phase can become more resistant well documented. Antibiotic resistance can be transmitted
to various environmental stresses (Kolter et al., 1993). among organisms by several mechanisms (Koutsoliout-
Feeds treated with organic acids are also believed to sou et al., 2001). Bacteria can become antibiotic resistant
limit pathogen infestation once ingested by the animal. by receiving and carrying antibiotic resistant encoding
Both formic and propionic acids have been reported in genes on a plasmid that is replicated as an extrachromo-
several studies to reduce Salmonella, coliforms, and Esche- somal genetic element during microbial multiplication
richia coli in the small intestinal, cecal, and fecal contents (Davies, 1994; Koutsolioutsou et al., 2001). In addition,
of chickens (Hinton and Linton, 1988; Rouse et al., 1988; mutations can occur on the bacterial chromosome that
Izat et al., 1990; McHan and Shotts, 1992). It is presumed decrease susceptibility to antibiotic killing mechanisms,
that the presence of organic acids in feeds limits pathogen or genes responsible for multiple resistances are turned
colonization by conversion of these acids into their respec- on by some sort of general stimuli (Nikaido, 1994; Spratt,
tive antibacterial forms once they have entered the diges- 1994; Miller and Sulavik, 1996; Alekshun and Levy, 1997).
tive tract of the animal consuming the treated feed. How- Although antibiotics have received the majority of the
ever, Hume et al. (1993a) observed inconsistent reduc- focus, other compounds used to control microbial con-
tions of Salmonella colonization in the crops and ceca of tamination in food production such as organic acids can
chickens infected with a Salmonella marker strain. Several also elicit resistance responses in microorganisms (Chap-
factors can alter the effective antibacterial concentration of man, 1998). Furthermore, the response of organisms to
an organic acid after the compound reaches the intestinal these compounds may decide whether or not they survive
tract. The pH of the gastrointestinal tract may be a factor a variety of environmental stressors encountered in ani-
that influences the level of dissociated organic acid pres- mal production and processing. Typical stresses encoun-
ent since the average pH levels increase toward neutrality tered in food systems by foodborne pathogens include
as the small intestine and large intestine are traversed in deficiencies in biologically available iron, heat and cold
most nonruminant animal species (Wolin, 1974; Watkins shock, oxidative stress, osmolarity stress, acid and alka-
and Miller, 1983). Likewise, the rumen in most ruminant line pH, and nutrient starvation (Archer, 1996). The ge-
species is generally buffered within a neutral pH range netic regulation of many of these environmental stressors
(Hungate, 1966; Wolin, 1974). In addition, some organic is becoming better understood and in many cases may
SYMPOSIUM: USE OF ANTIMICROBIALS IN PRODUCTION 635
involve highly intricate regulatory networks (Christman, mM) where they are widely used as preservatives due
et al., 1985; Spector et al., 1986; Matin et al., 1989; Spector to their antibacterial activities (Cherrington et al., 1991;
and Cubitt, 1992; Kolter et al., 1993; Foster and Spector, Hardin et al., 1995; Dorsa, 1997). The acid-tolerance sys-
1995). tem has been demonstrated to protect Salmonella against
Adding to this complexity is the recent suggestion that lethal effects of organic acids and increase survival in
E. coli possesses extracellular sensing components that fermented foods (Leyer and Johnson, 1992; Baik et al.,
may provide an early response to stress by the bacterial 1996). Although it might be anticipated that SCFA at
cell and result in induction of the corresponding stress acidic pH levels could induce acid tolerance, it has also
systems inside the bacterial cell (Rowbury, 2001). There been shown that high concentrations of SCFA at neutral
is some indication that if stress induction occurs in a pH are capable of inducing acid tolerance in E. coli and
bacterium when exposed to a single stressor, cross-protec- S. typhimurium (Kwon and Ricke, 1998a,b; Arnold et al.,
tion can be manifested across multiple stresses and gen- 2001). Kwon and Ricke (1998a,b) found that inorganic
eral tolerance of these environments is increased (Foster acid resistance of S. typhimurium was increased after expo-
and Spector, 1995a; Archer, 1996). Environmental changes sure to high concentrations of SCFA, and this SCFA-in-
may also potentially be relayed among members of bacte- duced acid resistance was further enhanced by acidic pH,
rial populations by cell-to-cell communication or quorum anaerobiosis, and prolonged exposure to SCFA. Previous
sensing of diffusible low molecular weight signal com- exposure to SCFA also increased S. typhimurium survivors

Downloaded from http://ps.oxfordjournals.org/ by bahram afsharmanesh on June 25, 2014


pounds (Surette et al., 1999; de Kievit and Iglewski, 2000; against extreme acid (pH 3.0), high osmolarity (2.5 M
Withers et al., 2001). NaCl), and reactive oxygen (20 mM hydrogen peroxide)
and consequently could potentially cross-protect Salmo-
Organic Acids and Induction nella from other environmental conditions that exist in
of Acid Tolerance food animal and processed food products (Kwon et al.,
2000).
Acid shock is a commonly occurring event experienced The interaction between acid shock and bacteria such
by foodborne pathogens during exposure to food pro- as Salmonella and E. coli, however, remains complicated
cessing and preservation. It has been known for over 60 because growth and survival responses to SCFA appear
yr that bacteria possess metabolic capabilities to respond to be dependent on not only the growth phase of the
to low external pH (Slonczewski, 1992). A more recent organism but atmospheric conditions (aerobic or anaero-
observation is that acid-sensitive bacteria could adapt to bic) used to recover survivors, salt form of the SCFA, and
acid stress by induction of an acid tolerance response. perhaps initial concentration of the SCFA (Kwon et al.,
This response essentially involves growth of the acid- 1998; Blankenhorn et al., 1999; Kwon and Ricke, 1999).
sensitive microorganism in a moderately low pH environ- Future research will require examining growth and sur-
ment that subsequently leads to survival when suddenly vival under more ecologically simulating conditions us-
exposed to what would normally be considered lethal ing techniques such as continuous culture to reproduce
acidic conditions (Foster, 1995). This adaptation permits the slow growth conditions and constant exposure to
the induction of genes involved in an acid-tolerance re- fermentation typically occurring in the gastrointestinal
sponse and synthesis of a series of acid shock proteins tract.
that are protective for extreme acidic conditions (Foster, In addition, genetic tools that allow for precise dissec-
1999). Multiple acid-tolerant systems that are physiologi- tion of all potential genes directly involved in SCFA and
cally distinct have been identified in foodborne pathogens stimulation of acid shock as well as peripheral systems
such as Salmonella and E. coli and are classified as log are required. Recent efforts with transposon mutagenesis
phase and stationary mechanisms (Foster, 1999). Foster and gene array approaches have indicated that these
(2001) has further classified these in ascending order of methods may be viable for identifying some of the pro-
tolerance as log phase bacteria, acid-adapted log phase spective genes involved in SCFA-induced acid tolerance
bacteria, stationary phase bacteria, and acid-adapted sta- (Arnold et al., 2001; Kwon et al., 2002).
tionary phase bacteria.
Weak organic acids such as the SCFA represent a poten- Organic Acids and Pathogenesis
tial acid stress exposure to foodborne pathogens due to of Foodborne Bacteria
high concentrations and low pH. Uncharged protonated
weak acids are freely diffusable across membranes with The concept of SCFA enhanced survivability to multi-
potential to lower internal pH (Bearson et al., 1997). It is ple environmental stresses has implications not only for
no surprise that an active area of research focus is the gastrointestinal ecology competitiveness of salmonellae
relationship between SCFA and potential for induction and foodborne pathogens in general, but, in the case of
of acid-tolerance mechanisms. High concentrations (any- SCFA, it raises practical issues regarding the use of or-
where from 10 to 100 mM) of SCFA produced by anaero- ganic acids as an intervention step in food processing.
bic native intestinal microflora are frequently encoun- Simply put this becomes a question of whether more
tered by foodborne pathogens (Meynell, 1963; Hungate, pathogenic subpopulations of a foodborne pathogen are
1966; Wolin, 1974; Cummings et al., 1987). Pathogens may selected for after exposure to organic acids. Microbial
also encounter the SCFA in food products (100 to 300 pathogenesis is a function of virulence expression in the
636 RICKE

organism and is essentially the degree to which organism enhance identification of signals for stimulation of viru-
can cause harm to its host. Microbial virulence phenotype lence in the pathogen of interest and provide specific
is defined as the ability of the respective pathogen to targets for development of more optimal strategies to
successfully infect the host and includes bacterial cell control this foodborne pathogen.
functions such as adherence to host tissue and invasion
into host cells (Mekalanos, 1992). It was originally sug- Conclusions
gested that because ingested foodborne pathogens come
in contact with low stomach pH, enhanced acid tolerance An obvious prerequisite for successful efforts to mini-
could increase the survivability after entry in the stomach mize the impact of antibiotic resistance in the animal
as well in vesicles of phagosomes after undergoing acidi- industry is development and implementation of alterna-
fication (Foster, 1992). In support of this concept, studies tive antimicrobials. Organic acids have a long history of
in which acid-sensitive mutants of pathogens have been use in the food industry as food preservatives and have
examined indicate that these mutants are also avirulent been demonstrated to be effective under a wide variety
(Lee et al., 1995; Wilmes-Riesenberg et al., 1997). Likewise, of food processing conditions. Although there is indica-
S. enteritidis isolates with enhanced acid tolerance were tion that some tolerance by foodborne pathogens is possi-
demonstrated to be more virulent in mice and more inva- ble, this tolerance may only be for particular circum-
sive in chickens (Humphrey et al., 1996). stances in which exposure to the organic acid is less than
maximum and provides the organism an opportunity to

Downloaded from http://ps.oxfordjournals.org/ by bahram afsharmanesh on June 25, 2014


Given that growth phase, acid tolerance, and virulence
may be linked, it is reasonable to suggest that SCFA may induce specific stress resistant and virulence mechanisms
also influence virulence (El-Gedaily et al., 1997). A series that allow for resistance to more severe acidic conditions.
of studies in which examination of the pathogenesis be- Improving effectiveness of organic acids will require
havior of S. typhimurium was determined as a function more understanding of general and specific stress re-
of adherence and invasion of mammalian cells after expo- sponse capabilities of foodborne pathogens. This would
sure to SCFA were done utilizing a tissue culture ap- include development and application of molecular tools
proach (HEp-2 cells; Gianella et al., 1973). Based on these for studying pathogen behavior in microbial ecosystems
studies, it appears that all SCFA (acetate, propionate, bu- in a variety of preharvest and postharvest food produc-
tyrate, and mixture of the three) are stimulatory to S. tion environments. Genetic techniques are now becoming
typhimurium invasion and adherence, but level of re- available to enable in depth dissection of the level of
sponse is dependent on growth phase, concentration, and interrelated regulation involved in the sensing and re-
pH (Durant et al., 1999, 2000c,d). sponse of a pathogen to the respective environment. How-
Although tissue culture approaches can help to identify ever, understanding the potentially high throughput of
which factors may be critical for adherence and invasion resulting data may present the next challenge to devising
phenotypes, quantifying relative importance and a regu- a practical strategy that anticipates potential problems
latory hierarchy is difficult. Understanding the genetics of and implementing effective control measures.
key regulatory genes in virulence expression in organisms
such as Salmonella has lead to the creation of bacterial ACKNOWLEDGMENTS
strains in which gene fusions have been constructed that
consist of the promoter region of the virulence regulatory This review was supported by Hatch grant H8311 ad-
gene hilA with a structural gene encoding for the biosyn- ministered by the Texas Agricultural Experiment Station
thesis of β-galactosidase (Bajaj et al., 1995). The resulting and USDA-NRI grant number 2002-35201-11608.
genetic fusion of these two gene components allows for
ease of measurement of hilA expression as a β-galactosi- REFERENCES
dase enzyme assay. The hilA gene is an ideal candidate Aarestrup, F. M., and H. C. Wegener. 1999. The effects of antibi-
because it encodes for a transcriptional regulator required otic usage in food animals on the development of antimicro-
for expression of genes encoding for proteins involved bial resistance of importance for humans in Campylobacter
in Salmonella pathogenicity and invasion of epithelial cells and Escherichia coli. Microbes Infect. 1:639–644.
Acuff, G. R., C. Vanderzant, J. W. Savell, D. K. Jones, D. B.
(Lee et al., 1992; Bajaj et al., 1995; Penheiter et al., 1997).
Griffin, and J. G. Ehlers. 1987. Effect of acid decontamination
When examining the influence using this approach, of beef subprimal cuts on the microbiological and sensory
growth phase appears to be an important interactive fac- characteristics of steaks. Meat Sci. 19:217–226.
tor based on Salmonella growth response and length of Alakomi, H.-L., E. Skyttä, M. Saarela, T. Mattila-Sandholm, K.
time for SCFA exposure prior to conducting the hilA gene Latva-Kala, and I. M. Helander. 2000. Lactic acid permeabil-
izes Gram-negative bacteria by disrupting the outer mem-
fusion assay (Durant et al., 2000a,b). brane. Appl. Environ. Microbiol. 66:2001–2005.
To further separate the complex interactions among Alekshun, M. N., and S. B. Levy. 1997. Regulation of chromo-
SCFA, other environmental stressors in food animal pro- somally mediated multiple antibiotic resistance: The mar re-
duction, pathogen virulence, and relevance to in vivo man- gulon. Antimicrob. Agents Chemother. 41:2067–2075.
ifestation of pathogenesis will require approaches such Archer, D. L. 1996. Preservation microbiology and safety: Evi-
dence that stress enhances virulence and triggers adaptive
as transposon mutagenesis to identify genes involved in mutations. Trends Food Sci. Technol. 7:91–95.
bacterial response to these harsh conditions (Kwon and Armstrong, D. G. 1984. Antibiotics as feed additives for rumi-
Ricke, 2000; Kwon et al., 2003). Such approaches will nant livestock. Pages 331–347 in Antimicrobials and Agricul-
SYMPOSIUM: USE OF ANTIMICROBIALS IN PRODUCTION 637
ture—Proceedings of the 4th International Symposium in Durant, J. A., D. E. Corrier, and S. C. Ricke. 2000a. Short chain
Agriculture: Benefits and Malefits. M. Woodbine, ed. But- volatile fatty acids modulate the expression of the hilA and
tersworths, London. invA genes of Salmonella typhimurium. J. Food Prot. 63:573–
Arnold, C. N., J. McElhanon, A. Lee, R. Leonhart, and D. A. 578.
Siegele. 2001. Global analysis of Escherichia coli gene expres- Durant, J. A. D. E. Corrier, L. H. Stanker, and S. C. Ricke.
sion during the acetate-induced tolerance response. J. Bacte- 2000b. Expression of the hilA Salmonella typhimurium gene in
riol. 183:2178–2186. a poultry Salmonella enteritidis isolate in response to lactate
Axe, D. D., and J. E. Bailey. 1995. Transport of lactate and acetate and nutrients. J. Appl. Microbiol. 89:63–69.
through the energized cytoplasmic membrane of Escherichia Durant, J. A., V. K. Lowry, D. J. Nisbet, L. H. Stanker, D. E.
coli. Biotechnol. Bioeng. 47:8–19. Corrier, and S. C. Ricke. 1999. Short-chain fatty acids affect
Baik, H. S., S. Bearson, S. Dunbar, and J. W. Foster. 1996. The cell-association and invasion of HEp-2 cells by Salmonella
acid tolerance response of Salmonella typhimurium provides typhimurium. J. Environ. Sci. Health B 34:1083–1099.
protection against organic acids. Microbiology 142:3195– Durant, J. A., V. K. Lowry, D. J. Nisbet, L. H. Stanker, D. E.
3200. Corrier, and S. C. Ricke. 2000c. Short-chain fatty acids alter
Bajaj, V., C. Hwang, and C. A. Lee. 1995. hilA is a novel ompR/ HEp-2 cell association and invasion by stationary growth
toxR family member that activates the expression of Salmo- phase Salmonella typhimurium. J. Food Sci. 65:1206–1209.
nella typhimurium invasion genes. Mol. Microbiol. 18:714–727. Durant, J. A., V. K. Lowry, D. J. Nisbet, L. H. Stanker, D. E.
Bearson, S., B. Bearson, and J. W. Foster. 1997. Acid stress re- Corrier, and S. C. Ricke. 2000d. Late logarithmic Salmonella
sponses in enterobacteria. FEMS Microbiol. Lett. 147:173–180. typhimurium HEp-2 cell association and invasion response to
Berchieri, A., Jr., and P. A. Barrow. 1996. Reduction in incidence short chain fatty acid addition. J. Food Safety 20:1–11.
of experimental fowl typhoid by incorporation of a commer- Eklund, T. 1983. The antimicrobial effect of dissociated and

Downloaded from http://ps.oxfordjournals.org/ by bahram afsharmanesh on June 25, 2014


cial formic acid preparation (Bio-Add) into poultry feed. undissociated sorbic acid at different pH levels. J. Appl. Bac-
Poult. Sci. 75:339–341. teriol. 54:383–389.
Blankenhorn, D., J. Phillips, and J. L. Slonczewski. 1999. Acid- Eklund, T. 1985. Inhibition of microbial growth at different pH
and base-induced proteins during aerobic and anaerobic levels by benzoic and propionic acids and esters of p-hydro-
growth of Escherichia coli revealed by two-dimensional gel xybenzoic acid. Int. J. Food Microbiol. 2:159–167.
electrophoresis. J. Bacteriol. 181:2209–2216. El-Gedaily, G. Paesold, C.-Y. Chen, D. G. Guiney, and M. Krause.
Carlson, V. L., and G. H. Snoeyenbos. 1970. Effect of moisture 1997. Plasmid virulence gene expression induced by short-
on salmonella populations in animal feeds. Poult. Sci. chain fatty acids in Salmonella dublin: Identification of rpoS-
49:717–725. dependent and rpoS-Independent mechanisms. J. Bacteriol.
Chapman, J. S. 1998. Characterizing bacterial resistance to pre- 179:1409–1412.
servatives and disinfectants. Int. Biodeter. & Biodegrad. Foster, J. W. 1992. Beyond pH homeostasis: The acid tolerance
41:241–245. response of salmonellae. Amer. Soc. Microbiol. News
Cherrington, C. A., M. Hinton, and I. Chopra. 1990. Effect of 58:266–270.
short-chain organic acids on macromolecular synthesis in Foster, J. W. 1995. Low pH adaptation and the acid tolerance
Escherichia coli. J. Bacteriol. 68:69–74. response of Salmonella typhimurium. Crit. Rev. Microbiol.
Cherrington, C. A., M. Hinton, G. C. Mead, and I. Chopra. 1991. 21:215–237.
Organic acids: Chemistry, antibacterial activity and practical Foster, J. W. 1999. When protons attack: Microbial strategies of
applications. Adv. Microb. Physiol. 32:87–108. acid adaptation. Curr. Opin. Microbiol. 2:170–174.
Christman, M. F., R. W. Morgan, F. S. Jacobson, and B. N. Foster, J. W. 2001. Acid stress responses of Salmonella and E.
Ames. 1985. Positive control of a regulon for defenses against coli: Survival mechanisms, regulation, and implications for
oxidative stress and some heat-shock proteins in Salmonella pathogenesis. J. Microbiol. 39:89–94.
typhimurium. Cell 41:753–762. Foster, J. W., and M. P. Spector. 1995. How Salmonella survive
Cummings, J. H., E. W. Pomare, W. J. Branch, C. P. E. Naylor, against the odds. Annu. Rev. Microbiol. 49:145–174.
and G. T. Macfarlane. 1987. Short chain fatty acids in human Freese, E., C. W. Sheu, and E. Galliers. 1973. Function of lipo-
large intestine, portal, hepatic and venous blood. Gut philic acids as antimicrobial food additives. Nature
28:1221–1227.
241:321–325.
Davidson, P. M. 2001. Chap. 29. Chemical preservatives and
Gianella, R. A., O. Washington, P. Gemski, and S. B. Formal.
natural antimicrobial compounds. Pages 593–627 in Food
1973. Invasion of HeLa cells by Salmonella typhimurium: A
Microbiology—Fundamentals and Frontiers. 2nd ed. M. P.
model for study of invasiveness of Salmonella. J. Infect. Dis.
Doyle, L. R. Beuchat, and T. J. Montville, ed. American Soci-
128:69–75.
ety for Microbiology, Washington, DC.
Glynn, M. K., C. Bopp, W. Dewitt, P. Dabney, M. Mokhtar,
Davies, J. 1994. Inactivation of antibiotics and the dissemination
and F. J. Angulo. 1998. Emergence of multidrug-resistant
of resistance genes. Science 264:375–382.
Davies, R. H., and C. Wray. 1996. Persistence of Salmonella enter- Salmonella enterica serotype Typhimurium DT104 infections
itidis in poultry units and poultry food. Br. Poult. Sci. in the United States. N. Engl. J. Med. 338:1333–1338.
37:589–596. Ha, S. D., K. G. Maciorowski, Y. M. Kwon, F. T. Jones, and S.
De Kievit, T. R., and B. H. Iglewski. 2000. Bacterial quorum C. Ricke. 1998a. Survivability of indigenous microflora and a
sensing in pathogenic relationships. Infect. Immun. Salmonella typhimurium marker strain in poultry mash treated
68:4839–4849. with buffered propionic acid. Anim. Feed Sci. Technol.
Denyer, S. P., and G. S. A. B. Stewart. 1998. Mechanisms of action 75:145–155.
of disinfectants. Int. Biodeterior. Biodegradation 41:261–268. Ha, S. D., K. G. Maciorowski, Y. M. Kwon, F. T. Jones, and S.
Dickson, J. S. 1992. Acetic acid action on beef tissue surfaces C. Ricke. 1998b. Indigenous feed microflora and Salmonella
contaminated with Salmonella typhimurium. J. Food Sci. typhimurium marker strain survival in poultry mash diets
57:297–301. containing varying levels of protein. Anim. Feed Sci. Technol.
Dixon, R. C., and P. B. Hamilton. 1981. Effect of feed ingredients 76:23–33.
on the antifungal activity of propionic acid. Poult. Sci. Hardin, M. D., G. R. Acuff, L. M. Lucia, J. S. Oman, and J. W.
60:2407–2411. Savell. 1995. Comparison of methods for decontamination
Dorsa, W. J. 1997. New and established carcass decontamination from beef carcass surfaces. J. Food Prot. 58:368–374.
procedures commonly used in the beef-processing industry. Heinzel, M. 1998. Phenomena of biocide resistance in microor-
J. Food Prot. 60:1146–1151. ganisms. Int. Biodeterior. Biodegradation 41:225–234.
638 RICKE
Hinton, M., and A. H. Linton. 1988. Control of Salmonella infec- a sustained acid tolerance response in virulent Salmonella
tions in broiler chickens by the acid treatment of their feed. typhimurium. Mol. Microbiol. 17:155–167.
Vet. Rec. 123:416–421. Levy, S. B., G. B. Fitzgerald, and A. B. Macone. 1976. Spread of
Hume, M. E., D. E. Corrier, S. Ambrus, A. Hinton Jr., and J. R. antibiotic-resistant plasmids from chicken to chicken and
DeLoach. 1993a. Effectiveness of dietary propionic acid in from chicken to man. Nature 260:40–42.
controlling Salmonella typhimurium colonization in broiler Leyer, G. J., and E. A. Johnson. 1992. Acid adaptation promotes
chicks. Avian Dis. 37:1051–1056. survival of Salmonella spp. in cheese. Appl. Environ. Micro-
Hume, M. E., D. E. Corrier, G. W. Ivie, and J. R. DeLoach. 1993b. biol. 58:2075–2080.
Metabolism of [14C] propionic acid in broiler chicks. Poult. Liu, T. S., G. H. Snoeyenbos, and V. L. Carlson. 1969. Thermal
Sci. 72:786–793. resistance of Salmonella senftenberg 775W in dry animal
Humphrey, T. J., and D. J. Lanning. 1988. The vertical transmis- feeds. Avian Dis. 13:611–631.
sion of salmonellas and formic acid treatment of chicken feed. Matin, A., E. A. Auger, P. H. Blum, and J. E. Schultz. 1989.
A possible strategy for control. Epidemiol. Infect. 100:43–49. Genetic basis of starvation survival in nondifferentiating bac-
Humphrey, T. J., A. Williams, K. McAlpine, M. S. Lever, J. teria. Annu. Rev. Microbiol. 43:293–316.
Guard-Petter, and J. M. Cox. 1996. Isolates of Salmonella enter- McHan, F., and E. B. Shotts. 1992. Effect of feeding selected short-
ica Enteritidis PT4 with enhanced heat and acid tolerance chain fatty acids on the in vivo attachment of Salmonella
are more virulent in mice and more invasive in chickens. typhimurium in chick ceca. Avian Dis. 36:139–142.
Epidemiol. Infect. 117:79–88. Mekalanos, J. J. 1992. Environmental signals controlling expres-
Hungate, R. E. 1966. The Rumen and Its Microbes. Academic sion of virulence determinants in bacteria. J. Bacteriol.
Press, New York. 174:1–7.
Izat, A. L., N. M. Tidwell, R. A. Thomas, M. A. Reiber, M. H. Meynell, G. G. 1963. Antibacterial mechanisms of the mouse

Downloaded from http://ps.oxfordjournals.org/ by bahram afsharmanesh on June 25, 2014


Adams, M. Colberg, and P. W. Waldroup. 1990. Effects of a gut II: The role of Eh and volatile fatty acids in the normal
buffered propionic acid in diets on the performance of broiler gut. Br. J. Exp. Pathol. 44:209–219.
chickens and on microflora of the intestine and carcass. Poult. Miller, P. F., and M. C. Sulavik. 1996. Overlaps and parallels
Sci. 69:818–826. in the regulation of intrinsic multiple-antibiotic resistance in
Juven, B. J., N. A. Cox, J. S. Bailey, J. E. Thomsen, O. W. Charles, Escherichia coli. Mol. Microbiol. 21:441–448.
and J. V. Shutze. 1984. Survival of Salmonella in dry food and Monnet, D. L. 1999. European recommendations to respond to
feed. J. Food Prot. 47:445–448. the threat of antimicrobial-resistant microorganisms. Am.
Khan, M., and M. Katamay. 1969. Antagonistic effect of fatty Soc. Microbiol. News 65:390–391.
acids against Salmonella in meat and bone meal. Appl. Micro- Mossel, D. A. A., and M. J. Koopman. 1965. Losses in viable
biol. 17:402–404. cells of salmonellae upon inoculation into dry animal feeds
Kolter, R., D. A. Siegele, and A. Tormo. 1993. The stationary of various types. Poult. Sci. 44:890–892.
phase of the bacterial life cycle. Annu. Rev. Microbiol. Neu, H. C. 1992. The crisis in antibiotic resistance. Science
47:855–874. 257:1064–1073.
Koutsolioutsou, A., E. A. Martins, D. G. White, S. B. Levy, and Nikaido, H. 1994. Prevention of drug access to bacterial targets:
B. Demple. 2001. A soxRS-constitutive mutation contributing Permeability barriers and active efflux. Science 264:382–388.
to antibiotic resistance in clinical isolate of Salmonella enterica Paster, N. 1979. A commercial scale study of the efficiency of
(Serovar Typhimurium). Antimicrob. Agents Chemother. propionic acid and calcium propionate as fungistats in poul-
45:38–43. try feed. Poult. Sci. 58:572–576.
Kwon, Y. M., and S. C. Ricke. 1998a. Induction of acid resistance Penheiter, K. L., N. Mahur, D. Giles, T. Fahlen, and B. D. Jones.
of Salmonella typhimurium by exposure to short-chain fatty 1997. Non-invasive Salmonella typhimurium mutants are avir-
acids. Appl. Environ. Microbiol. 64:3458–3463. ulent because of an inability to enter and destroy M cells of
Kwon, Y. M., and S. C. Ricke. 1998b. Survival of a Salmonella ileal Peyer’s patches Mol. Microbiol. 24:697–709.
typhimurium poultry isolate in the presence of propionic acid Rouse, J., A. Rolow, and C. E. Nelson. 1988. Research note:
under aerobic and anaerobic conditions. Anaerobe 4:251–256. Effect of chemical treatment of poultry feed on survival of
Kwon, Y. M., and S. C. Ricke. 1999. Salmonella typhimurium Salmonella. Poult. Sci. 67:1225–1228.
poultry isolate growth response to propionic acid and so-
Rowbury, R. J. 2001. Extracellular sensing components and ex-
dium propionate under aerobic and anaerobic conditions.
tracellular induction component alarmones give early warn-
Int. Biodeterior. Biodegradation 43:161–165.
ing against stress in Escherichia coli. Adv. Microb. Physiol.
Kwon, Y. M., S. D. Ha, and S. C. Ricke. 1998. Growth response
44:215–257.
of Salmonella typhimurium poultry isolate to propionic acid
Russell, J. B. 1992. Another explanation for the toxicity of fermen-
and sodium propionate under aerobic and anaerobic condi-
tation acids at low pH: Anion accumulation versus uncou-
tions. J. Food Safety 18:139–149.
pling. J. Appl. Bacteriol. 73:363–370.
Kwon, Y. M., L. F. Kubena, D. J. Nisbet, and S. C. Ricke. 2002.
Functional screening of bacterial genome for virulence genes Salmond, C. V., R. G. Kroll, and I. R. Booth. 1984. The effect of
by transposon footprinting. Methods Enzymol. 358:141–152. food preservatives on pH homeostasis in Escherichia coli. J.
Kwon, Y. M., S. Y. Park, S. G. Birkhold, and S. C. Ricke. 2000. Gen. Microbiol. 130:2845–2850.
Induction of resistance of Salmonella typhimurium to environ- Sheu, C. W., and E. Freese. 1972. Effects of fatty acids on growth
mental stresses by exposure to short-chain fatty acids. J. Food and envelope proteins of Bacillus subtilis. J. Bacteriol.
Sci. 65:1037–1040. 111:516–524.
Kwon, Y. M., and S. C. Ricke. 2000. Efficient amplification of Sheu, C. W., W. N. Konings, and E. Freese. 1972. Effects of
multiple transposon-flanking sequences. J. Microbiol. Meth- acetate and other short-chain fatty acids on sugars and amino
ods 41:195–199. acid uptake of Bacillus subtilis. J. Bacteriol. 111:525–530.
Lasko, D. R., N. Zamboni, and U. Sauer. 2000. Bacterial response Sheu, C. W., D. Salomon, J. L. Simmons, T. Sreevalsan, and E.
to acetate challenge: A comparison of tolerance among spe- Freese. 1975. Inhibitory effects of lipophilic acids and related
cies. Appl. Microbiol. Biotechnol. 54:243–247. compounds on bacteria and mammalian cells. Antimicrob.
Lee, C. A., B. D. Jones, and S. Falkow. 1992. Identification of a Agents Chemother. 7:349–363.
Salmonella typhimurium invasion locus by selection for hyper- Slonczewski, J. L. 1992. pH-regulated genes in enteric bacteria.
invasive mutants. Proc. Natl. Acad. Sci. USA 89:1847–1851. Am. Soc. Microbiol. News 58:140–144.
Lee, I. S., J. Lin, H. K. Hall, B. Bearson, and J. W. Foster. 1995. Smith, H. W. 1968. Anti-microbial drugs in animal feeds. Nature
The stationary-phase sigma factor σs (RpoS) is required for 218:728–731.
SYMPOSIUM: USE OF ANTIMICROBIALS IN PRODUCTION 639
Spector, M. P., Z. Aliabadi, T. Gonzalez, and J. W. Foster. 1986. Threlfall, E. J., L. R. Ward, J. A. Frost, and G. A. Willshaw. 2000.
Global control in Salmonella typhimurium: Two-dimensional The emergence and spread of antibiotic resistance in food-
electrophoretic analysis of starvation-, anaerobiosis-, and borne bacteria. Int. J. Food Microbiol. 62:1–5.
heat shock-inducible proteins. J. Bacteriol. 168:420–424. Vanderwal, P. 1979. Salmonella control of feedstuffs by pelleting
Spector, M. P., and C. L. Cubitt. 1992. Starvation-inducible loci or acid treatment. World’s Poult. Sci. J. 35:70–78.
of Salmonella typhimurium: Regulation and roles in starvation- Watkins, B. A., and B. F. Miller. 1983. Competitive gut exclusion
survival. Mol. Microbiol. 6:1467–1476. of avian pathogens by Lactobacillus acidophilus in gnotobiotic
Spratt, B. G. 1994. Resistance to antibiotics mediated by target chicks. Poult. Sci. 62:1772–1779.
alterations. Science 264:388–393. Williams, J. E. 1981. Salmonellas in poultry feed: A worldwide
Surette, M. G., M. B. Miller, and B. L. Bassler. 1999. Quorum review. Part III. Methods in control and elimination. World’s
sensing in Escherichia coli, Salmonella typhimurium and Vibrio Poult. Sci. J. 37:97–105.
harveyi: A new family of genes responsible for autoproducer Williams, J. E., and S. T. Benson. 1978. Survival of Salmonella
production. Proc. Natl. Acad. Sci. USA 96:1639–1644.
typhimurium in poultry feed and litter at three temperatures.
Tabib, Z., R. C. Dixon, W. M. Hagler, and P. B. Hamilton. 1984.
Avian Dis. 22:742–747.
Evidence for the inhibition of fungi in corn meal by organic
acids being dependent on the character of the corn meal. Wilmes-Riesenberg, M. R., J. W. Foster, and R. Curtiss III. 1997.
Poult. Sci. 63:1516–1523. An altered rpoS allele contributes to the avirulence of Salmo-
Thompson, J. L., and M. Hinton. 1997. Antibacterial activity of nella typhimurium LT2. Infect. Immun. 65:203–210.
formic and propionic acids in the diet of hens on salmonellas Withers, H., S. Swift, and P. Williams. 2001. Quorum sensing
in the crop. Br. Poult. Sci. 38:59–65. as an integral component of gene regulatory networks in
Thompson, J. L., and M. Hinton. 1996. Effect of short-chain fatty gram-negative bacteria. Curr. Opin. Microbiol. 4:186–193.

Downloaded from http://ps.oxfordjournals.org/ by bahram afsharmanesh on June 25, 2014


acids on the size of enteric bacteria. Lett. Appl. Microbiol. Wolin, M. J. 1974. Metabolic interactions among intestinal micro-
22:408–412. organisms. Am. J. Clin. Nutr. 27:1320–1328.

Das könnte Ihnen auch gefallen