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Prevalence and Biological Characteristics of Listeria Species Isolated from Livestock and Poultry Meat in Gansu Province, China

ZHIJIE DONG
ZHIJIE DONG
, 
YANAN SUN
YANAN SUN
, 
QING CAO
QING CAO
, 
HONGXIA LIU
HONGXIA LIU
, 
YUANYUAN LIU
YUANYUAN LIU
, 
QIHANG CAO
QIHANG CAO
, 
HUILIN WEI
HUILIN WEI
, 
CHEN SONG
CHEN SONG
, 
HUITIAN GOU
HUITIAN GOU
 et 
HUIWEN XUE
HUIWEN XUE
   | 24 mars 2023
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Article Category: ORIGINAL PAPER
Publié en ligne: 24 mars 2023
Pages: 11 - 20
Reçu: 06 oct. 2022
Accepté: 27 déc. 2022
DOI: https://doi.org/10.33073/pjm-2023-002
Mots clés
© 2023 ZHIJIE DONG et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1.

Distribution diagram of sampling points in Gansu province, China. The geographic locations of samples from slaughterhouses and markets. The five-pointed stars of red represent the different sampling sites.
Distribution diagram of sampling points in Gansu province, China. The geographic locations of samples from slaughterhouses and markets. The five-pointed stars of red represent the different sampling sites.

Fig. 2.

Serogroups by multiplex-PCR of L. monocytogenes isolates. M – DL 2000 DNA Maker, 1 – ATCC® 19115™ (serogroup 4b), 2 – ATCC® 19111™ (serogroup 1/2a), 3 – NCTC10890 (serogroup 1/2b), 4 – ATCC® 19112™ (serogroup 1/2c), 5–18 – the strains isolated in this study, 19 – L. innocua (negative control)
Serogroups by multiplex-PCR of L. monocytogenes isolates. M – DL 2000 DNA Maker, 1 – ATCC® 19115™ (serogroup 4b), 2 – ATCC® 19111™ (serogroup 1/2a), 3 – NCTC10890 (serogroup 1/2b), 4 – ATCC® 19112™ (serogroup 1/2c), 5–18 – the strains isolated in this study, 19 – L. innocua (negative control)

Fig. 3.

The growth curves of the Listeria monocytogenes isolates. Each data point is the average of triplicates, and the standard deviations are indicated as error bars. a) The growth curves for four serogroups of L. monocytogenes and one reference strain ATCC® 19111™, b) the growth curves for three isolates of L. innocua, c) the growth curves for three isolates of L. welshimeri.
The growth curves of the Listeria monocytogenes isolates. Each data point is the average of triplicates, and the standard deviations are indicated as error bars. a) The growth curves for four serogroups of L. monocytogenes and one reference strain ATCC® 19111™, b) the growth curves for three isolates of L. innocua, c) the growth curves for three isolates of L. welshimeri.

Fig. 4.

Biofilm formation by Listeria spp. isolates. The absorbance at 562 nm was measured for 22 L. innocua, 10 L. welshimeri, and 14 L. monocytogenes biofilms.
Biofilm formation by Listeria spp. isolates. The absorbance at 562 nm was measured for 22 L. innocua, 10 L. welshimeri, and 14 L. monocytogenes biofilms.

Fig. 5.

Cluster analysis of Listeria spp. isolates based on the sequences of the 16S rRNA genes. The tree was calculated using the Neighbor-Joining method. Isolates isolated in this study are indicated with circles (L. monocytogenes), boxes (L. innocua), and triangles (L. welshimeri). The scale bar represents 0.005 nucleotide substitutions per character. The numbers in the tree indicate the significance (percent of outcomes) of the branches (bootstrap analysis).
Cluster analysis of Listeria spp. isolates based on the sequences of the 16S rRNA genes. The tree was calculated using the Neighbor-Joining method. Isolates isolated in this study are indicated with circles (L. monocytogenes), boxes (L. innocua), and triangles (L. welshimeri). The scale bar represents 0.005 nucleotide substitutions per character. The numbers in the tree indicate the significance (percent of outcomes) of the branches (bootstrap analysis).

Primer information.

Gene target Primer sequence (5’-3’) Product size(bp) Application
prs-F GCTGAAGAGATTGCGAAAGAAG 370 Listeria species-specific primers
prs-R CAAAGAAACCTTGGATTTGCGG
Lmo2234-F TGTCCAGTTCCATTTTTAACT 420 Listeria monocytogenes specific primers
Lmo2234-R TTGTTGTTCTGCTGTACGA
Lin0464-F CGCATTTATCGCCAAAACTC 749 Listeria innocua specific primers
Lin0464-R TCGTGACATAGACGCGATTG
lmo0737-F AGGGCTTCAAGGACTTACCC 691 For the identification of Listeria monocytogenes serogroup
lmo0737-R ACGATTTCTGCTTGCCATTC
lmo1118-F AGGGGTCTTAAATCCTGGAA 909 For the identification of Listeria monocytogenes serogroup
lmo1118-R CGGCTTGTTCGGCATACTTA
ORF2819-F AGCAAAATGCCAAAACTCGT 471 For the identification of Listeria monocytogenes serogroup
ORF2819-R CATCACTAAAGCCTCCCATTG
ORF2110-F AGTGGACAATTGATTGGTGAA 597 For the identification of Listeria monocytogenes serogroup
ORF2110-R CATCCATCCCTTACTTTGGAC
16S rRNA-27F AGAGTTTGATCCTGGCTCAG 1,500 Used for cluster analysis
16S rRNA-1492R GGTTACCTTGTTACGACTT

Statistic of data on the prevalence of Listeria spp. according to regions and sample categories.

Number of samples Listeria monocytogenes positive samples (%) Listeria innocua positive samples (%) Listeria welshimeri positive samples (%) Listeria spp. positive samples (%)
Different regions
Lanzhou City 298 7 (2.4a) 30 (10.1b) 2 (0.7b) 39 (13.1b)
Qingyang City 275 6 (2.2b) 95 (34.6a) 4 (1.5a) 105 (38.2a)
Jiuquan City 129 1 (0.8c) 2 (1.6d) 0 (0c) 3 (2.3d)
Dingxi City 400 0 (0d) 6 (1.5d) 0 (0c) 6 (1.5e)
Zhangye City 285 0 (0d) 17 (6.0c) 4 (1.4a) 21 (7.4c)
Total             1,387 14 (1.0) 150 (10.8) 10 (0.7) 174 (12.6)
Different categories of samples
Pork 784 8 (1.0b) 81 (10.3d) 7 (0.9a) 96 (12.3c)
Beef and mutton 298 3 (1.0b) 35 (11.7b) 2 (0.7b) 40 (13.4a)
Chicken 264 3 (1.14a) 29 (11.0c) 1 (0.4c) 33 (12.5b)
Environment samples 41 0 (0b) 5 (12.2a) 0 (0d) 5 (12.2c)
Total             1,387 14 (1.0) 150 (10.8) 10 (0.7) 174 (12.6)

Result of drug susceptibility of Listeria spp. isolates.

Antibiotics Listeria monocytogenes Listeria innocua Listeria welshimeri
Resistant isolates Intermediate isolates Sensitive isolates Resistance rates (%) Resistant isolates Intermediate isolates Sensitive isolates Resistance rates (%) Resistant isolates Intermediate isolates Sensitive isolates Resistance rates (%)
Penicillin 13   0   1   92.9   2   0 20     9.1   3   0   7   30.0
Ofloxacin   1   2 11     7.1   1 13   8     4.6   2   0   8   20.0
Cefoxitin 14   0   0 100     0   0 22   0   2   0   8   20.0
Sulfamethoxazole 11   0   3   78.6 10   1 11   45.5   4   0   6   40.0
Tetracycline 14   0   0 100   14   0   8   63.6   4   0   6   40.0
Gentamicin   3   0 11   21.4   0   0 22   0   1   0   9   10.0
Streptomycin   2   1 11   14.3   5   0 17   23.7   1   0   9   10.0
Erythromycin 13   1   0   92.9   4   4 14   18.2   3   1   6   30.0
Acetylspiramycin 13   0   1   92.9   5   0 17   22.7   3   0   7   30.0
Fosfomycin   7   7   0   50.0 18   0   4   81.8   4   2   4   40.0
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Life Sciences, Microbiology and Virology
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