foods

Communication
Listeria monocytogenes in Fresh Produce: Outbreaks,
Prevalence and Contamination Levels
Qi Zhu, Ravi Gooneratne and Malik Altaf Hussain *
Department of Wine, Food and Molecular Biosciences, Lincoln University, Lincoln 7647, Canterbury,
New Zealand; [email protected] (Q.Z.); [email protected] (R.G.)
* Correspondence: [email protected]; Tel.: +64-220-144-552

Academic Editor: Christopher J. Smith

Received: 19 October 2016; Accepted: 8 March 2017; Published: 9 March 2017

Abstract: Listeria monocytogenes, a member of the genus Listeria, is widely distributed in agricultural
environments, such as soil, manure and water. This organism is a recognized foodborne pathogenic
bacterium that causes many diseases, from mild gastroenteritis to severe blood and/or central nervous
system infections, as well as abortion in pregnant women. Generally, processed ready-to-eat and
cold-stored meat and dairy products are considered high-risk foods for L. monocytogenes infections
that cause human illness (listeriosis). However, recently, several listeriosis outbreaks have been
linked to fresh produce contamination around the world. Additionally, many studies have detected
L. monocytogenes in fresh produce samples and even in some minimally processed vegetables.
Thus L. monocytogenes may contaminate fresh produce if present in the growing environment (soil and
water). Prevention of biofilm formation is an important control measure to reduce the prevalence and
survival of L. monocytogenes in growing environments and on fresh produce. This article specifically
focuses on fresh produce–associated listeriosis outbreaks, prevalence in growing environments,
contamination levels of fresh produce, and associated fresh produce safety challenges.

Keywords: Listeria monocytogenes; fresh produce; foodborne pathogen; contamination; listeriosis

1. Introduction
Listeria monocytogenes, a member of the genus Listeria, naturally occurs in agricultural
environments such as soil, manure and water [1]. Scientific literature frequently discusses the ability
of this microorganism to survive in the food-processing and produce-packing environment and
equipment [2]. It is a pathogenic bacterium that can cause a rare but dangerous infection called
listeriosis. The severity of listeriosis can range from mild gastroenteritis to severe disease conditions
(septicemia, encephalitis, meningitis, abortions and stillbirths) and can result in a high fatality rate in
immune-compromised populations [3]. Some people have a higher risk for developing listeriosis, such
as the elderly (>65 years) [4–6], infants and toddlers (<5 years) [4,7–9], pregnant women [3,8], and the
unborn [8,10]. About 17% of listeriosis cases occur in pregnant women [10]. According to the FDA
(Food and Drug Administration), about 2500 people suffer from listeriosis in the USA annually [10].
The mortality rate could be 20%–30% of those who contract listeriosis [3]. L. monocytogenes is responsible
for 19% of the total deaths due to the consumption of contaminated food in the USA [11].
L. monocytogenes is commonly found and isolated from processed, ready-to-eat (RTE) and
cold-stored meat and dairy products. An increasing number of recent reports show contamination
and prevalence of L. monocytogenes in fresh produce. L. monocytogenes bacteria have been
isolated from market or restaurant produce such as cabbage [12], corn [13], carrots [14–16],
lettuce [17–23], cucumbers [1,24,25], parsley [11,26,27] and salad vegetables [11,25,28,29]. Outbreaks
of L. monocytogenes infections associated with fresh produce have been reported in various parts of
the world [24]. For example, L. monocytogenes was responsible for the deaths of 10 people in a food

Foods 2017, 6, 21; doi:10.3390/foods6030021 www.mdpi.com/journal/foods

Foods 2017, 6, 21 2 of 11

poisoning listeriosis outbreak in chopped celery in Texas in 2010 [30]; in 2011, 30 people were infected
by listeria-contaminated melons in Colorado [31]; and in 2014, a listeria outbreak linked to caramel
apple contamination was reported in California [5]. This trend has continued and prevention of Listeria
contamination in fresh fruit and vegetables as well as fresh produce–associated listeriosis outbreaks is
now a food safety challenge.
This article focuses on fresh produce–associated listeriosis outbreaks, prevalence and survival of
L. monocytogenes in fresh produce growing environments, listeria contamination of produce and a brief
note on measures that could be used to control or reduce the level of contamination.

2. Foodborne Listeriosis
It has been over 90 years since human and animal listeriosis was first recognized as an infection
caused by a bacterium in the 1920s. The first conclusive link of L. monocytogenes to a foodborne outbreak
in 1981 stimulated research and survey work to determine the ubiquity of the organism and its method
of transmission [32]. L. monocytogenes gained recognition as a major foodborne pathogen when the
mortality rate did not diminish over the following years, even though the number of cases seemed to be
small compared with the estimated illnesses associated with salmonellosis and campylobacteriosis [33].
L. monocytogenes can cause two types of disease syndromes. Listeriosis is defined as being caused
by invasive L. monocytogenes. That is, the organism usually infects sterile parts of the body, such as the
liver [34], spleen [35], cerebral spinal fluid [36] and blood [37]. In healthy adults, diarrhea and fever
are the main symptoms [38], in pregnant women it is fever, diarrhea, abortion or stillbirth [39], and in
the newborn it can cause sepsis, pneumonia and meningitis [40–44]. L. monocytogenes can also cause a
non-invasive disease, usually as a febrile gastroenteritis or non-invasive gastroenteritis, and it has been
linked to outbreaks resulting from contaminated deli meat [45,46], chocolate milk [47], cheese [48–50],
smoked fish [51,52] and corn [13].
Foodborne listeriosis is a relatively rare but serious disease with a high fatality rate (up to 30%)
compared with diseases caused by other foodborne microbial pathogens [2,11]. The incidence rate
of foodborne outbreaks caused by contaminated fresh fruits and vegetables has shown an increasing
trend in recent years [53]. Most outbreaks have been reported in the USA, Europe, Canada, and to a
lesser extent in Australia and New Zealand [33].

3. Fresh Produce—Associated Listeriosis Outbreaks

In 1997, a serious Listeria outbreak associated with canned corn contaminated by L. monocytogenes
occurred in two primary schools and a university in Italy. The main symptoms in this outbreak
were febrile illness and gastroenteritis. A large number of people (2930 in total) developed febrile
gastroenteritis in these three institutes, including primary school students aged six to 10 years,
adult staff in the primary school, and students at the university. Investigation into this incident
showed that the symptoms occurred after eating food supplied by the same caterer. No other cases
were reported outside these three institutions in the same area during 1997 [13].
In 2010, the Texas Department of State Health Services (DSHS) reported a listeriosis outbreak
linked to chopped celery. Of the 10 infected patients aged 56 to 93 years admitted to the hospital,
five patients died within three months [30].
In 2011, an outbreak of listeriosis occurred in 28 different states in the US, caused by consumption
of contaminated melons, in which a total of 147 persons were affected and 33 died. In this outbreak,
an analysis of L. monocytogenes using pulsed-field gel electrophoresis (PFGE) matched the subtype
of L. monocytogenes colonies isolated from samples of cut cantaloupe and from the patients’ blood.
A pregnant woman who was affected in this outbreak had a miscarriage [54]. In the same year, another
outbreak associated with romaine lettuce was recorded across 19 states in the USA. In this outbreak,
84 became sick and of these, 15 died. The Federal Drug Agency (FDA) tested samples randomly
from the True Leaf Farms of California. The results of microbiological analyses were positive for
Foods 2017, 6, 21 3 of 11

L. monocytogenes. Approximately 30,000 pounds of chopped and bagged romaine lettuce in 90 cartons
were recalled [55].
A listeriosis outbreak associated with caramel apples occurred in December 2014 in the USA. The
vehicle for this outbreak was pre-packaged caramel apples. Testing confirmed that the origin of this
outbreak was from the firm’s apple-packing facility. In total, 35 people, including 11 pregnant women,
were infectedFoods
by2017, 6, 21
L. monocytogenes in 12 states. One of the infected pregnant women had 3 of 10
a miscarriage.
Seven out of were
the 35 people infected died during the outbreak [5]. More recently,
positive for L. monocytogenes. Approximately 30,000 pounds of chopped and bagged romaine a multistate outbreak
lettuce in 90 cartons were recalled [55].
of L. monocytogenes affected nine states in the USA in January 2016. Nineteen of the infected people
A listeriosis outbreak associated with caramel apples occurred in December 2014 in the USA.
were hospitalized, and one person from Michigan died of listeriosis. Epidemiological and laboratory
The vehicle for this outbreak was pre-packaged caramel apples. Testing confirmed that the origin of
evidence showedthis outbreakpackaged
that was from thesalads produced in
firm’s apple-packing Ohio
facility. In were
total, 35responsible for 11
people, including the outbreak. Table 1
pregnant
women, were infected by L. monocytogenes in 12 states. One of
below gives a summary of several outbreaks caused by fresh fruit and vegetables sincethe infected pregnant women had 1979.
a
miscarriage. Seven out of the 35 people infected died during the outbreak [5]. More recently, a
multistate outbreak of L. monocytogenes affected nine states in the USA in January 2016. Nineteen of
the infected Table Listeriosis
people1.were outbreaks
hospitalized, associated
and one person with
from fresh produce.
Michigan died of listeriosis.
Epidemiological and laboratory evidence showed that packaged salads produced in Ohio were
responsible for the outbreak. Table 1 below gives a summary of several outbreaks caused by fresh
Deaths/Cases
Outbreak Location/Year Food Vehicle References
fruit and vegetables since 1979.(% Mortality)
Boston, USA, 1979 3/20 (15) Raw vegetables Ho et al. [56]
Nova Scotia, Canada, 1981 Table 1. Listeriosis
17/41 (41) outbreaks associated with fresh
Vegetable produce.
mix for coleslaw Schlech et al. [57]
Moncalieri and Giaveno, Italy, 1997 0/2930 (0)
Deaths/Cases Corn Aureli et al. [13]
Outbreak
Texas, USA, 2010 Location/Year 5/10 (50) FoodChopped
Vehicle celery References Gaul et al. [30]
(% Mortality)
Colorado, USA, 2011
Boston, USA, 1979
33/147 (22)
3/20 (15)
Whole cantaloupes
Raw vegetables Ho et al. [56]
CDC [54]
Colorado,Nova
USA,Scotia,
2011 Canada, 1981 15/99 (15)
17/41 (41)
Lettuce
Vegetable mix for coleslaw
Shrivastava et al. [55]
Schlech et al. [57]
Illinois and Moncalieri
Michigan,and
USA, 2014
Giaveno, Italy, 1997
2/5 (40)
0/2930 (0)
Mung
Corn
bean sprouts Garner and Kathariou [58]
Aureli et al. [13]
California, USA, 2014
Texas, USA, 2010
1/32 (3)
5/10 (50)
Caramel apples
Chopped celery Gaul et al. [30]
CDC [5]
Ohio, USA, 2016
Colorado, USA, 2011
1/19 (5)
33/147 (22)
Packaged salads
Whole cantaloupes CDC [54]
CDC [50]
Colorado, USA, 2011 15/99 (15) Lettuce Shrivastava et al. [55]
Illinois and Michigan, USA, 2014 2/5 (40) Mung bean sprouts Garner and Kathariou [58]
4. Prevalence andCalifornia,
Survival USA,of L. monocytogenes
2014 1/32 (3) in Produce Growing Environments
Caramel apples CDC [5]
Ohio, USA, 2016 1/19 (5) Packaged salads CDC [50]
L. monocytogenes is present in many animals and humans [59,60], so it is possible to isolate
the bacterium 4. Prevalence
from theand Survival
feces of L. monocytogenes
of these sources and in Produce
in theirGrowing Environments
environment [61–63]. Moreover, fresh
produce and soilL.can monocytogenes is present in many
be contaminated animals and
by sewage humans
water [59,60],as
applied so it is possibleto
fertilizer to isolate
the crop the plants [64].
bacterium from the feces of these sources and in their environment [61–63]. Moreover, fresh produce
Thus, L. monocytogenes can be recycled among vegetables, humans and soils contaminated with feces
and soil can be contaminated by sewage water applied as fertilizer to the crop plants [64]. Thus, L.
(Figure 1). This bacterium
monocytogenes can behas an interesting
recycled life cycle
among vegetables, humansadaptation capability.
and soils contaminated It lives
with feces a saprophytic
(Figure
life in the soil1).but
Thiscan
bacterium
makehas theantransition
interesting life cyclea adaptation
into pathogeniccapability. It lives it
life when a saprophytic
enters into lifehuman
in the or animal
soil but can make the transition into a pathogenic life when it enters into human or animal cells [65].
cells [65]. The transition from a saprophyte to a cytosolic pathogen occurs through careful modulation
The transition from a saprophyte to a cytosolic pathogen occurs through careful modulation of the
of the activityactivity
of a specific regulatory
of a specific protein
regulatory protein (PrfA)
(PrfA) and
and the typethe type ofcarbon
of available available
source.carbon source.

Figure 1. Potential pathways of L. monocytogenes transmission to humans via fresh produce.

Figure 1. Potential pathways of L. monocytogenes transmission to humans via fresh produce.

L. monocytogenes has been isolated from RTE foods, such as freshly cut fruit [66] and fresh-cut
vegetables [67]. Additionally, L. monocytogenes has been isolated from the vegetable growing
environment [63]. Temperature, water activity (aw ) and the pH of foods are the main factors
Foods 2017, 6, 21 4 of 11

that influence the multiplication and survival of L. monocytogenes. Technical reports describe that
L. monocytogenes can grow under a wide range of growth conditions during food processing and storage,
for example, at temperatures as low as −0.4 ◦ C [68] and over a wide range of pH values from 4.3 to
9.4 [69]. In the case of the contaminated melons from Jensen Farms in Colorado (2011), the temperature
created an ideal environment for Listeria to grow. In addition, the equipment and machinery were
impossible to fully clean, and therefore had dirt on them. In addition, the potato washing machine was
used for washing cantaloupes. This resulted in the contamination of the cantaloupes. Furthermore,
trucks, including those used to haul rejected cantaloupes sent to cattle feedlots, were parked next to
the packing plant. This made it easy for the trucks to be contaminated with Listeria from the cattle
farms [54].
As mentioned above, many factors influence the prevalence of L. monocytogenes in fresh produce,
including direct or indirect contamination from the environment, such as from soil, water, compost and
feces (Table 2). In one research project, 174 samples were tested for L. monocytogenes and 48 produced
a positive reaction. All L. monocytogenes–positive water samples were from natural water sources
such as creek and pond water, and none of the 28 samples from piped water and well water tested
positive for L. monocytogenes [70]. A similar scenario was observed in an investigation into compost
and irrigated water [71]. Szymczak et al. [72] conducted research on the prevalence of L. monocytogenes
in fresh produce in relation to the type of soil, including those lands that were treated with natural
fertilizers, artificial fertilizers, and also wastelands and garden plots. It was apparent that the artificial
environment was more suitable for L. monocytogenes to survive. Exciting research on the factors
(including temperature and moisture) that can influence the survival of L. monocytogenes in soil was
carried out by McLaughlin et al. [73]. They used three marked colonies to monitor L. monocytogenes
survival in different soil types. They found that L. monocytogenes can survive in normal soil, and that
the bacterium preferred high-moisture-containing soils. In another research study, Locatelli et al. [74]
showed that physical and chemical properties of soil influence the survival of L. monocytogenes. Both
biotic and abiotic factors influence the survival of L. monocytogenes. So, it is quite clear that the external
environment (contaminated soil, water and nutrient content, soil properties) affects the survival of
L. monocytogenes. However, there could be other factors acting concurrently on L. monocytogenes
survival, especially under moist conditions.

Table 2. Prevalence of L. monocytogenes in a fresh produce growing environment.

Environment (Total Frequency a Number of

Country References
Number of Samples) Positive Samples (%)
Soil (178) 16 (9%)
Drag swab (175) 15 (9%)
Fecal (61) 9 (15%)
USA Strawn et al. [70]
Water (174) 48 (28%)
Engineered (28) 0 (0%)
Surface (146) 48 (33%)
Field 263 (17.5%)
USA Strawn et al. [71]
Water 74 (30%)
Poland Soil (1000) 55 (5.5%) Szymczak et al. [72]
Ireland Soil McLaughlin et al. [73]
French soil Locatelli et al. [74]
a Frequency data represents the number of positive samples (percent of positive samples).

5. L. monocytogenes Contamination Level of Fresh Produce

Contamination of fresh produce with pathogenic organisms affecting human health can occur at
the pre-harvest or post-harvest stage. There are numerous direct or indirect sources of contamination,
including animals or insects, soil, water, dirty equipment, and human handling. Many methods, such
Foods 2017, 6, 21 5 of 11

as the application of antimicrobial agents and UV radiation, have been used to reduce the microbial
load in fresh produce. However, a pathogenic bacterium such as L. monocytogenes might not be
completely inactivated due to its remarkable ability to survive in adverse conditions. In Table 3, several
studies are listed that illustrate the prevalence of L. monocytogenes in fresh produce. Szymczak et al. [72]
showed that 5% of parsley grown in naturally fertilized soil was positive for L. monocytogenes. In
addition, an assessment of lettuce for L. monocytogenes was undertaken from the farm to the table [18].
Results indicated that 1.05 log cfu/g L. monocytogenes were found in samples from restaurants and
0.146 log cfu/g in samples from homes. Although both these sets of samples had been treated before
cooking or eating, samples from home treatments were cleaner than those from restaurants [18]. Similar
studies showed that the washing of lettuce, cucumber and parsley markedly reduces the content of
L. monocytogenes [75]. They also studied the influence of the storage temperature, water temperature,
acetic acid concentration and immersion time on the survival of L. monocytogenes. As expected, the
higher storage temperatures increased the number of L. monocytogenes colonies. Although washing
with dilute acetic acid had some effect on reducing the number of L. monocytogenes, the extent of the
reduction depended largely on the structure of the vegetable [75]. It is speculated that washing fresh
produce to reduce the number of L. monocytogenes is more effective in fruits than it is in leafy vegetables.

Table 3. Some selected studies that reported the prevalence of L. monocytogenes in fresh produce.

Produce Country Prevalence a References

Vegetables China 140 (8, 5.7%) Wu et al. [76]
Poland 30 (3, 10.0%) Szymczak et al. [72]
Malaysia 16 (4, 25.0%) Ponniah et al. [11]
Parsley
Brazil 22 (1, 4.5%) Aparecida de Oliveira et al. [77]
Greece Nastou et al. [75]
Brazil 30 (1, 3.3%) Aparecida de Oliveira et al. [77]
Collard greens
Brazil 24 (1, 4.2%) Sant’Ana et al. [78]
Korea Ding et al. [18]
Brazil 152 (3, 2.0%) Sant’Ana et al. [78]
Lettuce
Nigeria Uzeh et al. [79]
Greece Nastou et al. [75]
Malaysia 32 (7, 21.9%) Ponniah et al. [11]
Cabbage Brazil 11 (2, 18.2%) Sant’Ana et al. [78]
Nigeria Uzeh et al. [79]
New Zealand Zhu et al. [80]
Spinach Brazil 11 (1, 9.1%) Sant’Ana et al. [78]
Carrot Malaysia 33 (8, 24.2%) Ponniah et al. [11]
Tomato Malaysia 32 (7, 21.9%) Ponniah et al. [11]
Malaysia 32 (7, 21.9%) Ponniah et al. [11]
Cucumber
Greece Nastou et al. [75]
Sprouts Korean 112 (1, 0.9%) Seo et al. [81]
a Number of total analyzed samples (number and percent of positive sample for L. monocytogenes).

A survey of L. monocytogenes contamination was published on minimally treated leafy vegetables,

including collard greens, cabbage, lettuce, Chinese cabbage, and arugula [77]. In total, this research
study examined 162 minimally processed leafy samples. Of these, only six samples were confirmed
for Listeria spp contamination and only three samples were confirmed as L. monocytogenes, and these
were found in collard greens, bunched parsley and spring onions. Research on market vegetables [78]
showed L. monocytogenes contamination in 3.1% of the samples. Five salad samples had counts between
1.0 × 101 and 2.6 × 102 cfu/g. Among the minimally processed vegetable samples evaluated in South
Korea, 0.3% of them tested positive in sprouts [81]. Uzeh et al. [79] tested many salad vegetables
Foods 2017, 6, 21 6 of 11

(lettuce, cabbages, carrots and cucumbers), and only cabbages and lettuce showed a positive reaction.
Thus, although L. monocytogenes levels may decrease after treatment, some colonies could still survive.

6. Prevention of Biofilm Formation to Reduce the Level of Contamination

Besides the factors associated with the growing environment, bacterial biofilm formation is an
important pathway for fresh produce contamination. Oliveira et al. [82] stated that the term biofilm
refers to a sessile form of microbial life, characterized by adhesion of microorganisms to biotic or
abiotic surfaces, with consequent production of extracellular polymeric substances.
Fresh produce comes into contact with many different kinds of surfaces at different temperatures
during processing or transport, and according to a study by Bonsaglia et al. [83], these two factors
influence the extent of L. monocytogenes biofilm formation. They compared L. monocytogenes biofilms
growing on three kinds of touched surfaces, polystyrene, glass and stainless steel, at three different
temperatures (4, 20 and 35 ◦ C). The results showed that L. monocytogenes attaches more easily
to hydrophilic surfaces (glass and stainless steel) than to hydrophobic surfaces (polystyrene).
Higher temperatures and longer incubation times decreased the extent of adherence to surfaces,
but the results were not significant.
Biofilms are produced by bacteria, including L. monocytogenes itself, to enhance their survival and
spread. Therefore, disrupting the biofilm of L. monocytogenes is a practical method to reduce its survival.
Botticella et al. [84] discussed the importance of biofilm formation in relation to the safety of fresh-cut
produce. According to them, biofilm formation allows L. monocytogenes to persist for long periods
of time in the food processing environment and thus represents a source of recurrent contamination
and poses a food safety risk. Results reported by Sant’Ana et al. [78] indicated that L. monocytogenes
persistence either in the field or in the processing environment of the tested RTE vegetables was due
to the presence of harborage sites due to biofilm formation. The most common methods employed
to reduce biofilm formation include physical (such as UV-C) and chemical (such as chlorine dioxide,
peroxyacetic acid) processes.
According to a recent study, physical methods are more effective in controlling biofilm formation
because of their minimal influence on product quality and stability [85]. These authors used
three physical methods to treat L. monocytogenes biofilms: 32 Hz ultra-sonication (US), 390 mJ/cm2
Ultraviolet-C (UV-C), and 750 mJ/cm2 cold oxygen plasma (COP). UV-C and COP were more effective
in reducing L. monocytogenes biofilm formation. Another effective method to reduce L. monocytogenes
biofilm production is to use organic acids combined with modified atmosphere packaging [86]. In that
study, by Bae et al. [86], cabbages were treated with 2% lactic acid for 10 min combined with modified
atmosphere packaging, and the number of L. monocytogenes were reduced by half (from 6.2 cfu/g
to 3.1 cfu/g). In addition, the modified atmosphere packaging (air, N2 gas, CO2 gas) proved to be
effective in delaying the growth of L. monocytogenes.

7. Conclusions
L. monocytogenes is widely present in agricultural production environments, and it is implicated
in the contamination of fresh crop produce. Most recent listeriosis outbreaks associated with fresh
produce are attributed to the crop growing environment, post-harvest processing and retailing. Several
reports have demonstrated that L. monocytogenes is commonly present in a wide variety of fresh produce
samples. It is important to reduce the level of this pathogen to enhance the fresh produce safety and
protect consumer health. Preventing L. monocytogenes biofilm formation through a practicable and
effective method will help to decrease its survival and contamination levels in fresh produce.

Acknowledgments: This study was carried out with the financial support of the Faculty of Agriculture and Life
Sciences, Lincoln University, Lincoln 7647, Canterbury, New Zealand.
Conflicts of Interest: The authors declare no conflict of interest.
Foods 2017, 6, 21 7 of 11

References
1. Jeyaletchumi, P.; Tunung, R.; Selina, P.M.; Chai, L.C.; Radu, S.; Farinazleen, M.G.; Cheah, Y.K.; Mitsuaki, N.;
Yoshitsugu, N.; Kumar, M.P. Assessment of Listeria monocytogenes in salad vegetables through kitchen
simulation study. J. Trop. Agric. Food Sci. 2012, 40, 55–62.
2. Azizoglu, R.A.; Gorski, L.; Kathariou, S. Listeria and produce: A troublesome liaison! Available online:
http://www.newfoodmagazine.com/advent-calendar/listeria-and-produce/ (accessed on 10 February
2017).
3. Swaminathan, B.; Gerner-Smidt, P. The epidemiology of human listeriosis. Microb. Infect. 2007, 9, 1236–1243.
[CrossRef] [PubMed]
4. Okutani, A.; Okada, Y.; Yamamoto, S.; Igimi, S. Nationwide survey of human Listeria monocytogenes infection
in Japan. Epidemiol. Infect. 2004, 132, 769–772. [CrossRef] [PubMed]
5. CDC. Multistate Outbreak of Listeriosis Linked to Commercially Produced, Prepackaged Caramel
Apples. 2015. Available online: http://www.cdc.gov/listeria/outbreaks/caramel-apples-12--14/index.html
(accessed on 15 October 2015).
6. Gillespie, I.A.; Mook, P.; Little, C.L.; Grant, K.; Adak, G.K. Listeria monocytogenes Infection in the Over-60s in
England Between 2005 and 2008: A Retrospective Case–Control Study Utilizing Market Research Panel Data.
Foodborne Pathog. Dis. 2010, 7, 1373–1379. [CrossRef] [PubMed]
7. Buzby, J.C. Children and Microbial Foodborne Illness. Food Rev. 2001, 24, 32.
8. Smith, B.; Kemp, M.; Ethelberg, S.; Schiellerup, P.; Bruun, B.G.; Gerner-Smidt, P.; Christensen, J.J.
Listeria monocytogenes: Maternal-foetal infections in Denmark 1994–2005. Scand. J. Infect. Dis. 2009, 41, 21–25.
[CrossRef] [PubMed]
9. Dilber, E.; Aksoy, A.; Çakir, M.; Bahat, E.; Kamaşak, T.; Dilber, B. Listeria monocytogenes meningitis in
two immunocompetent children. Ann. Trop. Paediatr. 2009, 29, 225–229. [CrossRef] [PubMed]
10. FDA. Food Safety for Pregnant Women. 2011. Available online: http://www.fda.gov/food/
foodborneillnesscontaminants/peopleatrisk/ucm312704.htm (accessed on 27 November 2015).
11. Scallan, E.; Hoekstra, R.M.; Angulo, F.J.; Tauxe, R.V.; Widdowson, M.A.; Roy, S.L.; Jones, J.L.; Griffin, P.M.
Foodborne illness acquired in the United States—Major pathogens. Emerg. Infect. Dis. 2011, 17, 7–15.
[CrossRef] [PubMed]
12. Ponniah, J.; Robin, T.; Paie, M.S.; Radu, S.; Ghazali, F.M.; Kqueen, C.Y.; Nishibuchi, M.; Nakaguchi, Y.;
Malakar, P.K. Listeria monocytogenes in raw salad vegetables sold at retail level in Malaysia. Food Cont. 2010,
21, 774–778. [CrossRef]
13. Aureli, P.; Fiorucci, G.C.; Caroli, D.; Marchiaro, G.; Novara, O.; Leone, L.; Salmaso, S. An Outbreak of Febrile
Gastroenteritis Associated with Corn Contaminated by Listeria monocytogenes. N. Engl. J. Med. 2000, 342,
1236–1241. [CrossRef] [PubMed]
14. Ruiz-Cruz, S.; Acedo-Félix, E.; Díaz-Cinco, M.; Islas-Osuna, M.A.; González-Aguilar, G.A. Efficacy of
sanitizers in reducing Escherichia coli O157:H7, Salmonella spp. and Listeria monocytogenes populations on
fresh-cut carrots. Food Cont. 2007, 18, 1383–1390. [CrossRef]
15. Sy, K.V.; Murray, M.B.; Harrison, M.D.; Beuchat, L.R. Evaluation of Gaseous Chlorine Dioxide as a Sanitizer
for Killing Salmonella, Escherichia coli O157:H7, Listeria monocytogenes, and Yeasts and Molds on Fresh and
Fresh-Cut Produce. J. Food Prot. 2005, 68, 1176–1187. [CrossRef] [PubMed]
16. Kamat, A.; Ghadge, N.; Ramamurthy, M.; Alur, M. Effect of low-dose irradiation on shelf life and
microbiological safety of sliced carrot. J. Sci. Food Agric. 2005, 85, 2213–2219. [CrossRef]
17. Thunberg, R.L.; Tran, T.T.; Bennett, R.W.; Matthews, R.N.; Belay, N. Microbial Evaluation of Selected Fresh
Produce Obtained at Retail Markets. J. Food Prot. 2002, 65, 677–682. [CrossRef] [PubMed]
18. Ding, T.; Iwahori, J.; Kasuga, F.; Wang, J.; Forghani, F.; Park, M.-S.; Oh, D.-H. Risk assessment for
Listeria monocytogenes on lettuce from farm to table in Korea. Food Control 2013, 30, 190–199. [CrossRef]
19. Althaus, D.; Hofer, E.; Corti, S.; Julmi, A.; Stephan, R. Bacteriological Survey of Ready-to-Eat Lettuce,
Fresh-Cut Fruit, and Sprouts Collected from the Swiss Market. J. Food Prot. 2012, 75, 1338–1341. [CrossRef]
[PubMed]
20. Francis, G.A.; O’Beirne, D. Isolation and Pulsed-Field Gel Electrophoresis Typing of Listeria monocytogenes
from Modified Atmosphere Packaged Fresh-Cut Vegetables Collected in Ireland. J. Food Prot. 2006, 69,
2524–2528. [CrossRef] [PubMed]
Foods 2017, 6, 21 8 of 11

21. Abadias, M.; Usall, J.; Anguera, M.; Solsona, C.; Viñas, I. Microbiological quality of fresh,
minimally-processed fruit and vegetables, and sprouts from retail establishments. Int. J. Food Microbiol. 2008,
123, 121–129. [CrossRef] [PubMed]
22. Soriano, J.M.; Rico, H.; Moltó, J.C.; Mañes, J. Listeria Species in Raw and Ready-to-Eat Foods from Restaurants.
J. Food Prot. 2001, 64, 551–553. [CrossRef] [PubMed]
23. Johannessen, G.S.; Loncarevic, S.; Kruse, H. Bacteriological analysis of fresh produce in Norway. Int. J.
Food Microbiol. 2002, 77, 199–204. [CrossRef]
24. Meldrum, R.J.; Little, C.L.; Sagoo, S.; Mithani, V.; McLauchlin, J.; de Pinna, E. Assessment of the
microbiological safety of salad vegetables and sauces from kebab take-away restaurants in the United
Kingdom. Food Microbiol. 2009, 26, 573–577. [CrossRef] [PubMed]
25. Jamali, H.; Paydar, M.; Chung, C.Y.; Wong, W.F. Prevalence of Listeria species and Listeria monocytogenes
serotypes in ready mayonnaise salads and salad vegetables in Iran. Afr. J. Microbiol. Res. 2013, 7, 1903–1906.
26. Gómez-Govea, M.; Solís-Soto, L.; Heredia, N.; García, S.; Moreno, G.; Tovar, O.; Isunza, G. Analysis of
microbial contamination levels of fruits and vegetables at retail in Monterrey, Mexico. J. Food Agric. Environ.
2012, 10, 152–156.
27. Öktem, A.B.; Bayram, G.; Ceylan, A.E.; Yentür, G. Prevalence of Listeria monocytogenes in Some Turkish
Foodstuffs. J. Food Qual. 2006, 29, 76–86. [CrossRef]
28. Easa, S.M.H. Microorganisms found in fast and traditional fast food. J. Am. Sci. 2010, 6, 515–537.
29. Skalina, L.; Nikolajeva, V. Growth potential of Listeria monocytogenes strains in mixed ready-to-eat salads.
Int. J. Food Microbiol. 2010, 144, 317–321. [CrossRef] [PubMed]
30. Gaul, L.K.; Farag, N.H.; Shim, T.; Kingsley, M.A.; Silk, B.J.; Hyytia-Trees, E. Hospital-Acquired Listeriosis
Outbreak Caused by Contaminated Diced Celery—Texas, 2010. Clin. Infect. Dis. 2013, 56, 20–26. [CrossRef]
[PubMed]
31. CDC. Multistate Outbreak of Listeriosis Linked to Whole Cantaloupes from Jensen Farms, Colorado.
2011. Available online: http://www.cdc.gov/listeria/outbreaks/cantaloupes-jensen-farms/ (accessed on
15 January 2016).
32. Liu, D. Handbook of Listeria monocytogenes; CRC Press: Boca Raton, FL, USA, 2008.
33. Todd, E.C.D.; Notermans, S. Surveillance of listeriosis and its causative pathogen, Listeria monocytogenes.
Food Cont. 2011, 22, 1484–1490. [CrossRef]
34. Wing, E.J.; Gregory, S.H. Listeria monocytogenes: Clinical and Experimental Update. J. Infect. Dis. 2002, 185,
S18–S24. [CrossRef] [PubMed]
35. Aoshi, T.; Carrero, J.A.; Konjufca, V.; Koide, Y.; Unanue, E.R.; Miller, M.J. The cellular niche of Listeria
monocytogenes infection changes rapidly in the spleen. Eur. J. Immunol. 2009, 39, 417–425. [CrossRef]
[PubMed]
36. Cone, L.A.; Leung, M.M.; Byrd, R.G.; Annunziata, G.M.; Lam, R.Y.; Herman, B.K. Multiple cerebral abscesses
because of Listeria monocytogenes: Three case reports and a literature review of supratentorial listerial brain
abscess(es). Surg. Neurol. 2003, 59, 320–328. [CrossRef]
37. Bhat, S.A.; Willayat, M.M.; Roy, S.S.; Bhat, M.A.; Shah, S.N.; Ahmed, A.; Maqbool, S.; Ganayi, B.A. Isolation,
molecular detection and antibiogram of Listeria monocytogenes from human clinical cases and fish of Kashmir,
India. Comp. Clin. Pathol. 2012, 22, 661–665. [CrossRef]
38. Salazar, J.K.; Wu, Z.; Yang, W.; Freitag, N.E.; Tortorello, M.L.; Wang, H.; Zhang, W. Roles of a Novel Crp/Fnr
Family Transcription Factor Lmo0753 in Soil Survival, Biofilm Production and Surface Attachment to Fresh
Produce of Listeria monocytogenes. PLoS ONE 2013, 8, e75736. [CrossRef] [PubMed]
39. Pérez-Trallero, E.; Zigorraga, C.; Artieda, J.; Alkorta, M.; Marimón, J.M. Two Outbreaks of Listeria
monocytogenes Infection, Northern Spain. Emerg. Infect. Dis. 2014, 20, 2155–2157. [CrossRef] [PubMed]
40. Khoury, N.T.; Hossain, M.M.; Wootton, S.H.; Salazar, L.; Hasbun, R. Meningitis With a Negative
Cerebrospinal Fluid Gram Stain in Adults: Risk Classification for an Adverse Clinical Outcome.
Mayo Clin. Proc. 2012, 87, 1181–1188. [CrossRef] [PubMed]
41. Brouwer, M.C.; van de Beek, D.; Heckenberg, S.G.B.; Spanjaard, L.; de Gans, J. Community-Acquired Listeria
monocytogenes Meningitis in Adults. Clin. Infect. Dis. 2006, 43, 1233–1238. [CrossRef] [PubMed]
42. Okike, I.O.; Lamont, R.F.; Heath, P.T. Do We Really Need to Worry About Listeria in Newborn Infants?
Pediatr. Infect. Dis. J. 2013, 32, 405–406. [CrossRef] [PubMed]
Foods 2017, 6, 21 9 of 11

43. Gaschignard, J.; Levy, C.; Romain, O.; Cohen, R.; Bingen, E.; Aujard, Y.; Boileau, P. Neonatal Bacterial
Meningitis: 444 Cases in 7 Years. Pediatr. Infect. Dis. J. 2011, 30, 212–217. [CrossRef] [PubMed]
44. Camacho-Gonzalez, A.; Spearman, P.W.; Stoll, B.J. Neonatal Infectious Diseases: Evaluation of Neonatal
Sepsis. Pediatr. Clin. North Am. 2013, 60, 367–389. [CrossRef] [PubMed]
45. Lin, C.-M.; Takeuchi, K.; Zhang, L.; Dohm, C.B.; Meyer, J.D.; Hall, P.A.; Doyle, M.P. Cross-Contamination
between Processing Equipment and Deli Meats by Listeria monocytogenes. J. Food Prot. 2006, 69, 71–79.
[CrossRef] [PubMed]
46. Gottlieb, S.L.; Newbern, E.C.; Griffin, P.M.; Graves, L.M.; Hoekstra, R.M.; Baker, N.L.; Hunter, S.B.; Holt, K.G.;
Ramsey, F.; Head, M.; et al. Multistate Outbreak of Listeriosis Linked to Turkey Deli Meat and Subsequent
Changes in US Regulatory Policy. Clin. Infect. Dis. 2006, 42, 29–36. [CrossRef] [PubMed]
47. Proctor, M.E.; Brosch, R.; Mellen, J.W.; Garrett, L.A.; Kaspar, C.W.; Luchansky, J.B. Use of pulsed-field gel
electrophoresis to link sporadic cases of invasive listeriosis with recalled chocolate milk. Appl. Environ.
Microbiol. 1995, 61, 3177–3179. [PubMed]
48. CDC. Multistate Outbreak of Listeriosis Linked to Roos Foods Dairy Products. Available online:
http://www.cdc.gov/listeria/outbreaks/cheese-02--14/index.html (accessed on 4 February 2015).
49. CDC. Listeriosis Infections Linked to Marte Brand Frescolina Ricotta Salata Cheese. Available online:
http://www.cdc.gov/listeria/outbreaks/cheese-09--12/index.html (accessed on 4 February 2015).
50. CDC. Multistate Outbreak of Listeriosis Linked to Packaged Salads Produced at Springfield, Ohio Dole
Processing Facility (Final Update). Available online: http://www.cdc.gov/listeria/outbreaks/bagged-
salads-01--16/ (accessed on 15 August 2016).
51. Tham, W.; Ericsson, H.; Loncarevic, S.; Unnerstad, H.; Danielsson-Tham, M.-L. Lessons from an outbreak of
listeriosis related to vacuum-packed gravad and cold-smoked fish. Int. J. Food Microbiol. 2000, 62, 173–175.
[CrossRef]
52. Rørvik, L.M. Listeria monocytogenes in the smoked salmon industry. Int. J. Food Microbiol. 2000, 62, 183–190.
[CrossRef]
53. Mukherjee, A.; Speh, D.; Jones, A.T.; Buesing, K.M.; Diez-Gonzalez, F. Longitudinal microbiological survey
of fresh produce grown by farmers in the upper Midwest. J. Food Prot. 2006, 69, 1928–1936. [CrossRef]
[PubMed]
54. CDC. Listeria outbreaks. Available online: https://www.cdc.gov/listeria/outbreaks/ (accessed on 25
September 2016).
55. Shrivastava, S. Listeria Outbreak—Bacteria Found in Romaine Lettuce: FDA. Available online: http://www.
ibtimes.com/listeria-outbreak-bacteria-found-romaine-lettuce-fda-320544 (accessed on 14 March 2016).
56. Ho, J.L.; Shands, K.N.; Friedland, G.; Eckind, P.; Fraser, D.W. An outbreak of type 4b Listeria monocytogenes
infection involving patients from eight boston hospitals. Arch. Intern. Med. 1986, 146, 520–524. [CrossRef]
[PubMed]
57. Schlech, W.F.; Lavigne, P.M.; Bortolussi, R.A.; Allen, A.C.; Haldane, E.V.; Wort, A.J.; Hightower, A.W.;
Johnson, S.E.; King, S.H.; Nicholls, E.S.; et al. Epidemic Listeriosis — Evidence for Transmission by Food.
N. Engl. J. Med. 1983, 308, 203–206. [CrossRef] [PubMed]
58. Garner, D.; Kathariou, S. Fresh produce-associated listeriosis outbreaks, sources of concern, teachable
moments, and insights. J. Food. Prot. 2016, 79, 337–344. [CrossRef] [PubMed]
59. Vela, A.I.; Fernandez-Garayzabal, J.F.; Vazquez, J.A.; Latre, M.V.; Blanco, M.M.; Moreno, M.A.; de la Fuente, L.;
Marco, J.; Franco, C.; Cepeda, A.; et al. Molecular Typing by Pulsed-Field Gel Electrophoresis of Spanish
Animal and Human Listeria monocytogenes Isolates. Appl. Environ. Microbiol. 2001, 67, 5840–5843. [CrossRef]
[PubMed]
60. Srinivasan, V.; Nam, H.M.; Nguyen, L.T.; Tamilselvam, B.; Murinda, S.E.; Oliver, S.P. Prevalence of
Antimicrobial Resistance Genes in Listeria monocytogenes Isolated from Dairy Farms. Foodborne Pathog.
Dis. 2005, 2, 201–211. [CrossRef] [PubMed]
61. Olier, M.; Pierre, F.; Lemaître, J.-P.; Divies, C.; Rousset, A.; Guzzo, J. Assessment of the pathogenic potential
of two Listeria monocytogenes human faecal carriage isolates. Microbiology 2002, 148, 1855–1862. [CrossRef]
[PubMed]
62. Nightingale, K.K.; Schukken, Y.H.; Nightingale, C.R.; Fortes, E.D.; Ho, A.J.; Her, Z.; Grohn, Y.T.;
McDonough, P.L.; Wiedmann, M. Ecology and Transmission of Listeria monocytogenes Infecting Ruminants
and in the Farm Environment. Appl. Environ. Microbiol. 2004, 70, 4458–4467. [CrossRef] [PubMed]
Foods 2017, 6, 21 10 of 11

63. Esteban, J.I.; Oporto, B.; Aduriz, G.; Juste, R.A.; Hurtado, A. Faecal shedding and strain diversity of
Listeria monocytogenes in healthy ruminants and swine in Northern Spain. BMC Vet. Res. 2009, 5, 2. [CrossRef]
[PubMed]
64. Food Safety Authority of Ireland. The Control and Management of Listeria monocytogenes Contamination of Food;
Food Safety Authority of Ireland: Dublin, Ireland, 2005.
65. Freitag, N.E.; Port, G.C.; Miner, M.D. Listeria monocytogenes—From saprophyte to intracellular pathogen.
Nat. Rev. Microbiol. 2009, 7, 623–628. [CrossRef] [PubMed]
66. Oliveira, M.; Viñas, I.; Colàs, P.; Anguera, M.; Usall, J.; Abadias, M. Effectiveness of a bacteriophage
in reducing Listeria monocytogenes on fresh-cut fruits and fruit juices. Food Microbiol. 2014, 38, 137–142.
[CrossRef] [PubMed]
67. Vandamm, J.P.; Li, D.; Harris, L.J.; Schaffner, D.W.; Danyluk, M.D. Fate of Escherichia coli O157:H7, Listeria
monocytogenes, and Salmonella on fresh-cut celery. Food Microbiol. 2013, 34, 151–157. [CrossRef] [PubMed]
68. Junttila, J.R.; Niemelä, S.I.; Hirn, J. Minimum growth temperature of Listeria monocytogenes and
non-haemolytic listeria. J. Appl. Bacteriol. 1988, 65, 321–327. [CrossRef] [PubMed]
69. Te Giffel, M.C.; Zwietering, M.H. Validation of predictive models describing the growth of Listeria
monocytogenes. Int. J. Food Microbiol. 1999, 46, 135–149. [CrossRef]
70. Strawn, L.K.; Fortes, E.D.; Bihn, E.A.; Nightingale, K.K.; Gröhn, Y.T.; Worobo, R.W.; Wiedmann, M.;
Bergholz, P.W. Landscape and Meteorological Factors Affecting Prevalence of Three Food-Borne Pathogens
in Fruit and Vegetable Farms. Appl. Environ. Microbiol. 2013, 79, 588–600. [CrossRef] [PubMed]
71. Strawn, L.K.; Gröhn, Y.T.; Warchocki, S.; Worobo, R.W.; Bihn, E.A.; Wiedmann, M. Risk Factors Associated
with Salmonella and Listeria monocytogenes Contamination of Produce Fields. Appl. Environ. Microbiol. 2013,
79, 7618–7627. [CrossRef] [PubMed]
72. Szymczak, B.; Szymczak, M.; Sawicki, W.; Dabrowski, W. Anthropogenic impact on the presence of
L. monocytogenes in soil, fruits, and vegetables. Folia Microbiol Praha 2014, 59, 23–29. [CrossRef] [PubMed]
73. McLaughlin, H.P.; Casey, P.G.; Cotter, J.; Gahan, C.G.M.; Hill, C. Factors affecting survival of Listeria
monocytogenes and Listeria innocua in soil samples. Arch. Microbiol. 2011, 193, 775–785. [CrossRef] [PubMed]
74. Locatelli, A.; Spor, A.; Jolivet, C.; Piveteau, P.; Hartmann, A. Biotic and Abiotic Soil Properties Influence
Survival of Listeria monocytogenes in Soil. PLoS ONE 2013, 8, e75969. [CrossRef] [PubMed]
75. Nastou, A.; Rhoades, J.; Smirniotis, P.; Makri, I.; Kontominas, M.; Likotrafiti, E. Efficacy of household
washing treatments for the control of Listeria monocytogenes on salad vegetables. Int. J. Food Microbiol. 2012,
159, 247–253. [CrossRef] [PubMed]
76. Wu, S.; W, Q.; Zhang, J.; Chen, M.; Yan, Z.; Hu, H. Listeria monocytogenes prevalence and characteristics in
retails raw foods in China. PLoS ONE 2015, 10, e0136682. [CrossRef] [PubMed]
77. Aparecida, O.M.; Abeid Ribeiro, E.G.; Morato Bergamini, A.M.; Pereira De Martinis, E.C. Quantification
of Listeria monocytogenes in minimally processed leafy vegetables using a combined method based on
enrichment and 16S rRNA real-time PCR. Food Microbiol. 2010, 27, 19–23. [CrossRef] [PubMed]
78. Sant’Ana, A.S.; Igarashi, M.C.; Landgraf, M.; Destro, M.T.; Franco, B.D.G.M. Prevalence, populations and
pheno- and genotypic characteristics of Listeria monocytogenes isolated from ready-to-eat vegetables marketed
in São Paulo, Brazil. Int. J. Food Microbiol. 2012, 155, 1–9. [CrossRef] [PubMed]
79. Uzeh, R.; Adepoju, A. Incidence and survival of Escherichia coli O157: H7 and Listeria monocytogenes on salad
vegetables. Int. Food Res. J. 2013, 20, 1921–1925.
80. Zhu, Q.; Hussain, M.A. Prevalence of Listeria species in fresh salad vegetables and ready-to-eat foods
containing fresh produce marketed in canterbury, New Zealand. Adv. Food Technol. Nutr. Sci. Open J. 2014, 1,
5–9. [CrossRef]
81. Seo, Y.-H.; Jang, J.-H.; Moon, K.-D. Microbial evaluation of minimally processed vegetables and sprouts
produced in Seoul, Korea. Food Sci. Biotechnol. 2010, 19, 1283–1288. [CrossRef]
82. de Oliveira, M.M.M.; Brugnera, D.F.; Alves, E.; Piccoli, R.H. Biofilm formation by Listeria monocytogenes on
stainless steel surface and biotransfer potential. Braz. J. Microbiol. 2010, 41, 97–106. [CrossRef] [PubMed]
83. Bonsaglia, E.C.R.; Silva, N.C.C.; Fernades Júnior, A.; Araújo Júnior, J.P.; Tsunemi, M.H.; Rall, V.L.M.
Production of biofilm by Listeria monocytogenes in different materials and temperatures. Food Cont. 2014, 35,
386–391. [CrossRef]
Foods 2017, 6, 21 11 of 11

84. Botticella, G.; Russo, R.; Capozzi, V.; Amado, M.L.; Massa, S.; Spano, G.; Beneduce, L. Listeria monocytogenes,
biofilm formation and fresh cut produce. 2013. Available online: http://www.formatex.info/microbiology4/
vol1/114-123.pdf (accessed on 5 February 2017).
85. Srey, S.; Park, S.Y.; Jahid, I.K.; Ha, S.-D. Reduction effect of the selected chemical and physical treatments to
reduce L. monocytogenes biofilms formed on lettuce and cabbage. Food Res. Int. 2014, 62, 484–491. [CrossRef]
86. Bae, Y.-M.; Choi, N.-Y.; Heu, S.; Kang, D.-H.; Lee, S.-Y. Inhibitory effects of organic acids combined with
modified atmosphere packaging on foodborne pathogens on cabbage. J. Korean Soc. Appl. Biol. Chem. 2011,
54, 993–997. [CrossRef]

© 2017 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC BY) license (http://creativecommons.org/licenses/by/4.0/).