Food Control 85 (2018) 269e275

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Food Control
journal homepage: www.elsevier.com/locate/foodcont

Prevalence and characterization of Salmonella serovars isolated from

farm products in Shanghai
Pei’en Ni a, Qian Xu a, Yujie Yin a, Danlei Liu a, Jvmei Zhang b, Qingping Wu b, Peng Tian c,
Xianming Shi a, Dapeng Wang a, *
a
MOST-USDA Joint Research Center for Food Safety, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China
b
State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application,
Guangdong Open Laboratory of Applied Microbiology, Guangdong Institute of Microbiology, Guangzhou 510070, China
c
Produce Safety and Microbiology Research Unit, Western Regional Research Center, Agricultural Research Service e United States Department of
Agriculture, Albany, CA 94706, USA

a r t i c l e i n f o a b s t r a c t

Article history: Salmonella is a major bacterial pathogen that causes foodborne illness worldwide. Farm products,
Received 12 June 2017 including raw meat, vegetables, and dairy products, may be important vehicles for transmitting food-
Received in revised form borne pathogens, thus representing a threat to human health. In this study, the prevalence of Salmonella
9 October 2017
isolated from farm products in Shanghai was investigated and characterized. A total of 344 samples were
Accepted 10 October 2017
randomly collected from farmers’ markets, supermarkets, milk factories, and a slaughterhouse in
Available online 11 October 2017
Shanghai between March 2016 and February 2017. Using the Chinese National Standard method (GB
4789.4e2010), 83 (24.1%) samples were determined to be positive for Salmonella, including 42 (42.4%)
Keywords:
Salmonella
pork, 21 (41.2%) chicken, 15 (31.3%) beef, 3 (16.7%) mutton, 1 (4.5%) lettuce, and 1 (3.8%) cucumber. All egg
Farm products and raw milk samples were negative for Salmonella. Ninety strains were isolated from the positive
MLST samples, and 12 serovars were identified, including Enteritidis (n ¼ 35), Typhimurium (n ¼ 22), Derby
PFGE (n ¼ 13), Anatum (n ¼ 5), Agona (n ¼ 4), Thompson (n ¼ 3), Indiana (n ¼ 3), Braenderup (n ¼ 1),
Heidelberg (n ¼ 1), Infantis (n ¼ 1), Litchfield (n ¼ 1) and Manhattan (n ¼ 1). All isolates were char-
acterized by multilocus sequence typing (MLST) and subtyped by pulsed-field gel electrophoresis (PFGE).
Thirteen sequence types (ST) were identified, among which ST11, ST34, and ST40 were the predominant
STs. ST was highly correlated with serovar type. The results of PFGE showed that 73 PFGE patterns were
generated from the isolates. In conclusion, Salmonella contamination was common among farm products
in Shanghai, and the diversity of the pathogen was high among the isolates.
© 2017 Elsevier Ltd. All rights reserved.

1. Introduction Shanghai is one of the largest cities in China, with a population

A large number of farm products, including meat, vegetable, and
dairy products are consumed every day worldwide. Such products
are critical vehicles for foodborne pathogens, including Salmonella,
Staphylococcus aureus and norovirus (Abdalrahman, Wells, & Fakhr,
2015; Crump, Sjo €lund-Karlsson, Gordon, & Parry, 2015; Robilotti,
Deresinski, & Pinsky, 2015; Xue & Zhang, 2013). Salmonella is the
leading pathogenic bacteria causing foodborne diseases (EFSA,
2010). It has been reported that 93.8 million cases of gastroenter-
itis are caused by Salmonella species globally every year (Majowicz
et al., 2010).
* Corresponding author.
E-mail addresses: dapengwang@sjtu.edu.cn, norovirus@163.com (D. Wang).
of more than 24 million. Due to the high demand for farm products,
a large number of farm products are sourced from other cities (Yu
et al., 2015). Non-typhoidal Salmonella infection in children with
acute gastroenteritis is reportedly common in Shanghai (Li et al.,
2014). However, less information is available regarding the preva-
lence and diversity of this pathogen among farm products from the
largest import city. There have been a few studies reporting
contamination with Salmonella in other cities in China. Bai et al.
(2015) reported that the detection rates of Salmonella in chicken
and pork collected in Henan, China, were 45.2% and 29.2% respec-
tively. Yang et al. (2011) reported that the prevalence of Salmonella
in raw poultry at the retail level in six provinces and two metro-
politan cities in China ranged from 38.9% to 65.3%. Zhao et al. (2016)
examined the prevalence of Salmonella isolated from free-range

https://doi.org/10.1016/j.foodcont.2017.10.009
0956-7135/© 2017 Elsevier Ltd. All rights reserved.
270 P. Ni et al. / Food Control 85 (2018) 269e275
chickens in Shandong province, China, reporting a detection rate of
12.7%. Finally, the prevalence of Salmonella contamination in raw
chicken carcasses at the retail level in China was reported by Zhu
et al. (2014). They found that the overall Salmonella contamina-
tion rate was 41.6%. Most these studies were focused on chicken
and pork products. Reports of Salmonella contamination in other
meat products such as beef and mutton, as well as vegetable, dairy
and egg products in China are limited. Therefore, in this study, we
assessed Salmonella contamination among common meat products
(chicken, pork, beef and mutton), common ready-to-eat vegetables
(romaine lettuce and cucumber), raw milk, and eggs collected in
Shanghai, China.
Molecular typing is a useful method for distinguishing among
different bacterial isolates and can be used to trace the origins of
pathogenic bacteria (Clark et al., 2012). Multilocus sequence typing
(MLST) and pulsed-field gel electrophoresis (PFGE) have been
widely-used to type bacteria (Acar et al., 2017; Chang et al., 2016).
MLST involves a PCR assay using primers that are specific for
different housekeeping loci, followed by sequencing of the result-
ing gene fragments (Larsen et al., 2012; Maiden et al., 1998), which
is used to efficiently analyze the diversity and evolution of bacteria.
PFGE can be used to analyze not only genomic DNA but also plasmid
DNA in bacteria. PFGE is widely recognized as the gold standard
typing method owing to its high discriminatory power (Gatto et al.,
2006). Therefore, these two methods have been used to distinguish
bacterial isolates at the nucleic acid level in previous studies (Cai
et al., 2016; Yang et al., 2016; Zhou et al., 2017).
In this study, 8 types of farm products (pork, chicken, beef,
mutton, romaine lettuce, cucumber, eggs and raw milk) were
randomly sampled between March 2016 and February 2017 from
nine different districts in Shanghai. Eight of the nine districts are
suburbs that together comprise most of Shanghai, while the ninth,
Xuhui, is metropolitan area. The prevalence, serotype distribution,
and genetic diversity of Salmonella were analyzed, and molecular
typing was performed using MLST and PFGE.
2. Materials and methods
2.1. Sample collection
A total of 344 samples were collected every 2 weeks between
March 2016 and February 2017 in Shanghai, including pork fillets
(n ¼ 99), chicken breasts (n ¼ 51), beef tendons (n ¼ 48), mutton
chops (n ¼ 18), eggs (n ¼ 225 in 45 samples), romaine lettuce
(n ¼ 22), cucumber (n ¼ 26), and raw milk (n ¼ 35). The samples
were randomly selected from supermarkets (n ¼ 183), farmers’
markets (n ¼ 109), a slaughterhouse (n ¼ 17) and dairy farms
(n ¼ 35) located within the nine districts of Shanghai. The nine
districts included the following: Minhang, Fengxian, Jinshan,
Songjiang, Qingpu, Pudong, Jiading, Baoshan and Xuhui. Samples
from each district consisted of all eight types of farm products. After
collection, all samples were placed in sterilized containers, trans-
ported to our laboratory within 2 h in portable cooler containing
ice, and processed immediately upon arrival.
2.2. Isolation and identification of Salmonella
Salmonella was isolated from samples using the Chinese Na-
tional Standard method (GB 4789.4e2010). Briefly, each sample
(25.0 g or 25.0 mL) was homogenized with 225.0 mL of buffered
peptone water (BPW; Land Bridge Technology, Beijing, China) by a
homogenizer (AES Chemunex, Marcy-l’Etoile, France) for pre-
enrichment in a sterile polythene bag (Whirl-Park®, Nasco,
Shenzhen, China).
After incubation at 37  C for 18 h, 1.0 mL pre-enriched culture
was inoculated into 10.0 mL tetrathionate broth base (TTB; Land
Bridge Technology) or 10.0 mL selenite cystine broth (SC; Land
Bridge Technology) and incubated at 42  C or 37  C, respectively.
After 24 h of incubation, a loop from each broth culture was
streaked onto bismuth sulfite agar (BS; Land Bridge Technology)
plates or xylose lysine deoxycholate medium (XLD; Land Bridge
Technology) plates and incubated at 37  C for 48 h or 24 h,
respectively.
Next, 3e4 typical colonies (black colonies with a black zone and
metallic sheen surrounding the colony on BS agar; red colonies
with black centers on XLD agar) were selected from plates and
confirmed by polymerase chain reaction (PCR) assays using primers
invAF (50 -GTGAAATTATCGCCACGTTCGGGCA-30 ) and invAR (50 -
TCATCGCACCGTCAAAGGAACC-30 ) (Wang et al., 2012). PCR was
performed in a 25.0 mL mixture containing 12.5 mL of 2  Taq Master
Mix (Vazyme Nanjing, China), 9.5 mL ddH2O, 1.0 mL of sample DNA,
and 1.0 mL of each primer. All strains were serotyped by slide
agglutination using commercial O and H antisera (Tianrun Bio-
Pharmaceutical, Ningbo, China) according to the Kauffmann-
White scheme.
2.3. DNA extraction
Genomic DNA was extracted with a TIANamp Bacterial DNA Kit
(Tiangen, Beijing, China) according to the manufacturer’s protocol.
2.4. MLST
Seven housekeeping genes (aroC, dnaN, hemD, hisD, purE, sucA,
and thrA) were selected for the molecular typing of Salmonella
isolates according to the guidelines for MLST from the University of
Warwick (http://mlst.warwick.ac.uk/mlst/). The recommended
protocols from the MLST homepage were used, including
primers and PCR conditions (http://mlst.warwick.ac.uk/mlst/dbs/
Senterica). After amplification, PCR products were sequenced by
GENEWIZ (Suzhou, China). Sequences and strain information was
submitted to the MLST database to assign allelic type (AT) and
sequence type (ST) numbers.
2.5. Pulsed-field gel electrophoresis (PFGE)
PFGE was performed according to the protocol developed by the
Centers for Disease Control and Prevention (CDC) with minor
modifications (Ribot et al., 2006). Briefly, isolates were incubated
on Luria-Bertani (LB) plates at 37  C for 24 h, and Salmonella cells
were washed and suspended in cell suspension buffer (CSB). Then,
20.0 mL of proteinase K (20.0 mg/mL) was added to 400.0 mL of cell
suspension. Next, 400.0 mL of melted 1.0% SeaKem Gold agarose
(Lonza, Morristown, NJ, USA) with 1.0% sodium dodecyl sulfate
(SDS) was mixed with 400.0 mL of cell suspension. The mixture was
immediately poured into the plug molds, cooled, and transferred to
a lysis solution. The agarose-embedded DNA was lysed for 2 h and
then stored in 0.5  Tris-Borate-EDTA (TBE) at 4  C. The bacterial
cells in the agarose plugs were digested with 50 U of XbaI (TaKaRa,
Dalian, China) for 2 h at 37  C. Digested fragments were resolved in
1.0% SeaKem Gold agarose gel in 0.5  TBE using a Chef Mapper
electrophoresis system (Bio-Rad, Hercules, CA, USA). Electropho-
resis was performed at 14  C for 19 h. After staining with Gel-Rad,
DNA bands were obtained by UV trans-illumination (Bio-Rad).
Fingerprinting profiles were analyzed using BioNumerics Software
(Applied Maths, Kortrijk, Belgium). According to the manufacturer’s
instructions, clonal relationships among strains were determined
using the Dice coefficient, and a dendrogram was generated using
the unweighted pair-group method (UPGMA) with settings of 0.5%
optimization and 1.5% position tolerance.
 P. Ni et al. / Food Control 85 (2018) 269e275 271

2.6. Statistical analysis Table 2

The prevalence of Salmonella in different sample groups.

IBM SPSS Statistics Software (version 19; Armonk, NY, USA) was Meat Vegetable Raw milk Egg Total
used for statistical analysis. The chi-square test was used to Total number 216 48 35 45 344
compare differences, which were considered significant at p < 0.05. Positive number 81 2 0 0 83
Positive rate (%) 37.5 4.16 0 0 24.1
3. Results

3.1. Prevalence and identification of Salmonella in farm products Table 3

The prevalence of Salmonella in different seasons.

A total of 344 farm products were divided into four groups:
meat, vegetable, dairy and egg products (Table 2). There were 216
meat samples, including pork (n ¼ 99), chicken (n ¼ 51), beef
(n ¼ 48), and mutton (n ¼ 18); 48 vegetable samples, including
romaine lettuce (n ¼ 22) and cucumbers (n ¼ 26); 35 raw milk
samples; and 45 egg samples (Table 1). Based on our analysis, 90
Salmonella strains were isolated from 83 (24.1%) of the 344 samples.
Prevalence rates for Salmonella were 37.5% and 4.16% among meat
and vegetable samples, respectively. No Salmonella was detected
among the egg or raw milk samples (Table 2). The prevalence in the
meat group (37.5%) was significantly higher than those in the other
groups (p ¼ 0.0007). There were no significant differences in
detection rates among different meat types, except for in mutton,
where the detection rate was significantly lower (p ¼ 0.038). There
was no significant difference in the detection rate among samples
collected from supermarkets and those collected from farmers’
markets (p ¼ 0.92). However, the detection rate from the slaugh-
terhouse (52.9%) was significantly higher than those from super-
markets or farmers’ markets (p ¼ 0.014).
The detection rate was higher in the summer (30.3%) than
during other seasons. As shown in Table 3, the prevalence of Sal-
monella in the summer was significantly higher than that in
autumn (p ¼ 0.043), although there were no significant differences
in detection rates among the other three seasons.
3.2. Distribution of Salmonella serovars
Among the 90 isolates, 12 serotypes were recovered, including
Enteritidis (n ¼ 35), Typhimurium (n ¼ 22), Derby (n ¼ 13), Anatum
(n ¼ 5), Agona (n ¼ 4), Thompson (n ¼ 3), Indiana (n ¼ 3), Braen-
derup (n ¼ 1), Heidelberg (n ¼ 1), Infantis (n ¼ 1), Litchfield (n ¼ 1)
and Manhattan (n ¼ 1) (Table 4). Enteritidis was the most prevalent
serovar detected in all samples types except for mutton, in which
Typhimurium was the only serovar isolated. Eight serovars were
identified among pork samples, and Derby (23.9%) and Typhimu-
rium (23.9%) were tied for the second most prevalent serovars in
these samples. Eight serovars were identified among chicken
samples, and the second and third most prevalent serovars were
Typhimurium (25.0%) and Thompson (8.3%), respectively. Other
serovars, including Braenderup, Indiana, Litchfield and Manhattan
were uncommon. Five Salmonella serovars were recovered from
beef, including Enteritidis (53.3%), Agona (20.0%), Typhimurium
(13.3%), Thompson (6.7%) and Derby (6.7%). Among the other
samples, including lettuce and cucumber, Enteritidis was the only
Season Positive rate (%) Total number Positive number
Spring (2016.3e2016.5) 29.8ab 47 14
Summer (2016.6e2016.8) 30.3a 89 27
Autumn (2016.9e2016.11) 18.3b 120 22
Winter (2016.12e2017.2) 22.7ab 88 20
Different letters in the same column indicate a statistical difference (p < 0.05).
serovar identified.
3.3. MLST analysis
Through MLST analysis, the 90 strains were classified into 13 STs.
ST11 (n ¼ 35), ST34 (n ¼ 21) and ST40 (n ¼ 13) were the most
popular STs among the 90 isolates. The remaining isolates belonged
to ST13, ST17, ST18, ST19, ST22, ST26, ST32, ST64, ST214 and ST322.
There was a strong correlation between ST and serovar (Table 5).
Each serovar represented a unique ST except for S. Typhimurium
isolates, which were resolved into ST19 and ST34 in this study.
Almost all 13 STs were found in the Minhang, Jinshan, and
Fengxian districts. ST11 and ST34 were found in seven different
districts, followed by ST40 and ST64 which were found in three
districts and one slaughterhouse (Fig. 1). Other STs were distributed
sporadically among the sampling sites.
3.4. PFGE analysis
A total of 90 Salmonella strains were analyzed by PFGE with XbaI
enzyme digestion, with 73 PFGE patterns (PF1ePF73) identified
among the 90 strains. The PFGE patterns were grouped into nine
clusters (AeI) (Fig. 2). The most common cluster was cluster E,
which included 34 strains. Although these 34 isolates were
collected from different areas and at different times of the year,
these isolates were highly similar genetically. For example, the PF53
and PF54 isolates, which were obtained from different districts,
exhibited high similarity. Similar results were observed in other
clusters. Most isolates from the same serovar were grouped into the
same cluster; however, some isolates of the same serovar belonged
to different clusters. For example, two S. Derby (202 and 205) iso-
lates were found in cluster A while the other S. Derby isolates found
in cluster E. S. Enteritidis isolates were distributed across almost all
clusters. Some isolates from different districts shared the same
PFGE patterns. For example, PF14 included two S. Typhimurium
isolates from Xuhui, one S. Typhimurium isolate from Minhang and
one S. Typhimurium isolate from Jinshan.

Table 1
Isolation of Salmonella from different farm products.

Sample Source

Pork Chicken Beef Mutton Lettuce Cucumber Raw milk Egg Supermarket Free market Slaughterhouse Dairy farm

Total number 99 51 48 18 22 26 35 45 183 109 17 35

Positive number 42 21 15 3 1 1 0 0 46 28 9 0
Positive rate (%) 42.4a 41.2a 31.3a 16.7b 4.5 3.8 0 0 25.1 25.7 52.9 0

Different letters in the same row indicate a statistical difference (p < 0.05).
272 P. Ni et al. / Food Control 85 (2018) 269e275

Table 4
Distribution of Salmonella serovars from different samples.

serovar No. of isolates Type of sample

Pork Chicken Beef Mutton Lettuce Cucumber Raw Milk Egg

Enteritidis 35 14 11 8 1 1
Typhimurium 22 11 6 2 3
Derby 13 11 1 1
Anatum 5 5
Agona 4 1 3
Thompson 3 2 1
Indiana 3 2 1
Braenderup 1 1
Heidelberg 1 1
Infantis 1 1
Litchfield 1 1
Manhattan 1 1

Table 5
Prevalence of sequence types (STs) for the Salmonella isolates.

STs Serovars Allelic type No. of isolates

aroC dnaN hemD hisD purE sucA thrA

ST11 Enteritidis 5 2 3 7 6 6 11 35
ST34 Typhimurium 10 19 12 9 5 9 2 21
ST19 Typhimurium 10 7 12 9 5 9 2 1
ST40 Derby 19 20 3 20 5 22 22 13
ST64 Anatum 10 14 15 31 25 20 33 5
ST26 Thompson 14 13 18 12 14 18 1 3
ST13 Agona 3 3 7 4 3 3 7 4
ST17 Indiana 8 8 11 11 5 11 15 3
ST22 Braenderup 12 2 15 14 11 14 16 1
ST322 Heidelberg 43 47 49 49 41 15 114 1
ST32 Infantis 17 18 22 17 5 21 19 1
ST214 Litchfield 14 72 21 12 6 19 15 1
ST18 Manhattan 9 9 6 12 9 12 2 1

million people consume a large number of farm products daily.

Fig. 1. Minimum spanning tree analysis of 90 Salmonella isolates collected from
different sampling sites in Shanghai. The size of each circle indicates the quantity of
STs. Different sampling cites are represented by different colors, and the size of each
color block represents the number of STs in that area.
4. Discussion
Shanghai is the economic and financial center of China. Over 24
Therefore, it is critical to understand contamination levels among
farm products in Shanghai. In this study, 344 samples were
randomly collected from nine districts in Shanghai over the course
of a year. The prevalence of Salmonella was found to be significantly
different among the different types of samples.
In this study, 24.1% of farm products were positive for Salmo-
nella, including pork, beef, chicken, mutton, egg, lettuce, cucumber,
and raw milk products. This rate is higher than that reported for
Changchun, China (16.8%; Ren et al., 2016), but lower than rates
reported for other areas of China. In Henan, China for example,
36.1% of poultry meat was found to be positive for Salmonella in
2007 (Cui et al., 2009). In Shaanxi, China, 44.0% of retail meat
samples, including pork, beef, chicken, and lamb products, were
positive for Salmonella between 2007 and 2008 (Yang et al., 2010).
These differences in contamination levels may reflect true differ-
ences in prevalence among different geographic locations used for
sampling. In other countries, lower contamination levels have been
reported for farm products, including raw meat, minced meat,
burger samples, raw eggs and raw milk from Gondar, Ethiopia
(5.5%; Ejo, Garedew, Alebachew, & Worku, 2016) and chicken from
Ireland (2.5%; Duggan et al., 2012). The prevalence of Salmonella
among meat products (37.5%) in our study was much higher than
that among vegetables (4.2%), which is consistent with the findings
of our previous study (Chen, Zhang, Yang, Wu, & Xu, 2013). In
addition, no Salmonella-positive samples were recovered from eggs
in our study, which is consistent with a recent report from the
United States CDC that showed that the percentage of outbreaks
associated with eggs has decreased in the U.S. (Gould et al., 2013).
All eggs collected for this study were from commercial-scale egg
 P. Ni et al. / Food Control 85 (2018) 269e275
Fig. 2. Dendrogram of PFGE patterns of 90 Salmonella isolates. PF, PFGE pattern; S/F/P,
supermarket, farmers’ market, pig slaughterhouse, respectively.
farms. With the development of new technologies, mechanized
production is now used in most egg farms, which may reduce
contamination levels of Salmonella among eggs. According to a
previous report, Duan et al. (2012) found that the prevalence of
Salmonella on egg farms was quite low due to the advanced disease
control practice used in commercial-scale farms. Wang et al. (2012)
reported the prevalence of Salmonella isolates from commercial
eggs from Shaanxi province, China to be 2.7%. Similarly, we did not
observe any Salmonella contamination of the raw milk tested,
which is consistent with a report showing no Salmonella detected
among 247 raw milk samples collected from Shandong province,
China from 2003 to 2010 (Shao, 2011). Some previous studies have
identified a high level of S. aureus contamination in raw milk (Chen
et al., 2003; Jamali, Paydar, Radmehr, Ismail, & Dadrasnia, 2015).
However, S. aureus contamination in raw milk was not investigated
273
in our study.
Our data (Table 1) indicated that samples from the slaughter-
house had the highest detection rate (52.9%) relative to others.
Compared with the contamination rate (21.2%) observed among
samples from pig slaughterhouses reported by Bai et al. (2015), the
detection rate in our study was much higher, which was likely due
to the time at which sampling occurred. In this study, all samples
from the pig slaughterhouse were collected in September, and the
temperature and moisture conditions for that month were suitable
for the growth of Salmonella. This represents a significant risk, as
Salmonella from these products may be easily transferred to other
products.
In this study, contamination with Salmonella was affected by the
season in which sampling occurred, with the detection rate in
summer being the highest. Similar results were reported in previ-
ous studies (Edrington et al., 2008; Hald, Wingstrand, Swanenburg,
Von Altrock, & Thorberg, 2003). This may be because in the sum-
mer months in Shanghai (JuneeAugust), the high humidity and
temperature are conducive to the growth of Salmonella. In contrast,
the detection rate in autumn was lower than that in winter, which
differs from the results of a previous study (Zhu et al., 2014). One
reason for this is that all raw milk samples were collected in
autumn; the low contamination level among raw milk samples
thus resulted in a decrease in the overall detection rate in autumn.
We identified a total of 12 serovars in this study (Table 4), among
which S. Enteritidis (38.9%) and S. Typhimurium (24.4%) were the
most common. Our results were consistent with epidemic and
clinical data in Shanghai (Zhang et al., 2014). In all meats except
mutton, the most common serovar was Enteritidis, followed by
Typhimurium, and Derby (Table 4). These results are consistent
with those of previous reports (Wang et al., 2015; Zhang et al.,
2009; Zhao et al., 2016). In contrast, only Typhimurium was
detected in mutton in our study (Table 4). In a previous study, Yin
et al. (2016) reported that 10 serovars were found in mutton,
with the predominant serotype being Hadar, among samples from
Uighur, Xinjiang, China. This conflicting result may reflect differ-
ences in the geographical locations and customs. Mutton is a major
meat product in Xinjiang, and its consumption is more common in
this area than in Shanghai. Due to the different customs and sea-
sons in Shanghai, local residents consume mutton in autumn and
winter, which may have an effect on the prevalence of Salmonella
serotypes detected. Our results suggest that S. Enteritidis, S.
Typhimurium, and S. Derby should be considered major targets for
surveillance and control.
In this study, 90 Salmonella isolates were characterized by MLST,
and a total of 13 STs were identified (Table 5). ST11 was the most
common, especially among chicken samples, which is in agreement
with the results of a previous study (Zhao et al., 2016). Among pork
samples, ST11, ST40, and ST34 were the dominant STs. These three
major STs were also commonly recovered from all nine districts,
indicating that they are widespread in Shanghai and may pose a
significant threat to human health. However, ST11 was not found in
the slaughterhouse, despite being very common among pork
samples in this study. Considering that only one pig slaughterhouse
was included in this study, other pig slaughterhouses in Shanghai
or in other areas that supply pork products for Shanghai may be the
source of these ST11 isolates. Other issues such as cross-
contamination in the supply chain may also explain this phenom-
enon. The same ST was found in both retail markets and the
slaughterhouse, suggesting that these isolates derived from the
same contamination source. Most STs corresponded to unique
serovars; for example, ST11, ST34 and ST40 corresponded to the
serovars Enteritidis, Typhimurium, and Derby, respectively, in this
study. However, ST19 and ST34 both corresponded with S. Typhi-
murium, and a similar result was found in other study (Achtman
274 P. Ni et al. / Food Control 85 (2018) 269e275
et al., 2012). These results suggest that STs and serovars are strongly
correlated, which is in accordance with previous reports (Ben-Darif
et al., 2010; Ranieri, Shi, Switt, den Bakker, & Wiedmann, 2013).
PFGE is a useful tool for determining genetic variability among
Salmonella isolates. In this study, the 90 Salmonella isolates
exhibited a large number of PFGE patterns due to their high di-
versity (Fig. 2). However, similar PFGE pattern were observed across
different districts, farm products, and sampling times. For example,
two isolates (112.1, isolated from chicken, and 212.1, isolated from
beef) were collected from a supermarket and a farmers’ market in
different districts and different sampling times but shared the same
PFGE pattern (PF14) and the same ST (ST34). In addition, four S.
Typhimurium isolates recovered from different farm products at
different times and from different districts shared the same PFGE
pattern (PF14). Four isolates with the pattern PF56 were all
collected from Minhang at different times, suggesting the persis-
tence of a specific strain in that area over the sampling period.
These results suggest the possibility that specific Salmonella strains
may persist or be transmitted between different farm products over
the course of the year. Among pork samples, isolates collected from
retail markets and the slaughterhouse exhibited high genotype
similarity. For example, strain 46.1 and strain 199, which were
collected from a supermarket and the slaughterhouse, respectively,
shared identical PFGE pattern (PF4) and the same ST (ST17). This
suggests that the slaughterhouse may be the source of Salmonella
contamination among pork products in Shanghai. Cross-
contamination could also occur at the retail level. Strain 118.1 iso-
lated from pork and strain 120 isolated from beef were recovered at
the same time and at the same location. They shared the PF53
pattern according to the dendrogram of PFGE patterns, suggesting
the possibility of cross-contamination at the retail level, as these
samples did not share their production chains. Compared with
MLST, PFGE demonstrated more power for molecular typing. For
example, the 90 Salmonella isolates were divided into 73 PFGE
patterns and only 13 STs, and isolates with the same ST did not
necessarily exhibit the same PFGE pattern. However, MLST analysis
may be a useful complement to serotype research.
In conclusion, the results of this study indicated that Salmonella
contamination is common among farm products from the nine
districts of Shanghai. Due to their relatively high contamination
levels, the Enteritidis, Typhimurium, and Derby serovars should
receive greater attention in the future. By combining MLST and
PFGE subtyping, the Salmonella isolates were found to be geneti-
cally diverse, and the temporal persistence of Salmonella was
observed over the sampling period. In retail markets, cross-
contamination between farm products may be a considerable
problem, and greater attention should be paid to this issue. For
consumers, fully cooked farm products may present lower health
risks, as Salmonella has a low tolerance for heating. The data ob-
tained in this study may be useful for tracing possible foodborne
outbreaks of Salmonella and may aid in the development of sur-
veillance programs for farm products.
Conflict of interest
None declared.
Acknowledgments
This work was jointly supported by the Ministry of Science and
Technology of China (2016YFE0106100), the Shanghai Agriculture
Applied Technology Development Program (Grant No. G
20150405), and the National Nature Science Foundation of China
(31601562).
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