Scientia Agricultura Sinica ›› 2019, Vol. 52 ›› Issue (9): 1646-1656.doi: 10.3864/j.issn.0578-1752.2019.09.015

• ANIMAL SCIENCE·VETERINARY SCIENCE·RESOURCE INSECT • Previous Articles     Next Articles

Investigation on the Antibiotic Resistance of Staphylococcus Methicillin-Resistant MLSB from Food Animals in Six Provinces of China

LI ShuMin1,FANG LiangXing1,LI Liang2,ZHAO Meng1,LU Xiao1,GU WeiQi1,LIAO XiaoPing1,SUN Jian1,XIONG YanQiong2,LIU YaHong1()   

  1. 1 College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China
    2 LABioMed at Harbor-UCLA Medical Center, Torrance, CA 90502, USA
  • Received:2018-05-17 Accepted:2019-03-06 Online:2019-05-01 Published:2019-05-16
  • Contact: YaHong LIU E-mail:gale@scau.edu.cn

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Abstract:

【Objective】 The aim of this study was to investigate the prevalence of genes encoding resistance to macrolides, lincosamides and streptogramins (MLSB) among methicillin-resistant Staphylococcus aureus from food animals in China. 【Method】Staphylococcus aureus were isolated from about 6 500 samples of pig, chicken and duck origins among six different provinces, including Guangdong, Henan, Hebei, Fujian and so on, in China. LA-MRSA was confirmed from staphylococcus aureus through detecting the susceptibility of oxacillin and the presence of mecA and mecC genes. Susceptibility test of LA-MRSA resistant to MLSB and other common antibiotics were determined by agar dilution method and microdilution broth method. And the MLSB resistance genes (ermA-C and ereA-B) and other common resistant genes were detected by PCR. 【Result】Four hundred and eighty Staphylococcus aureus isolates and 101 LA-MRSA isolates were found among about 6 500 samples, and the isolation rate of Staphylococcus aureus and the detection rate of LA-MRSA among Staphylococcus aureus isolates were 1.54% (101/6500) and 21.04% (101/480), respectively. Susceptibility test showed high resistance rate of LA-MRSA to MLSB was found with over 99.00% among LA-MRSA isolates. High resistance rate to other common antibiotics was also found among LA-MRSA, including florfenicol, tetracycline, cefotaxime and gentamycin with the resistance rates of 99.01%, 96.39%, 96.39%, 96.39% and 94.05%, respectively. MLSB resistance genes detection showed that ermC was found in all LA-MRSA isolates, followed by ereB (79.20%), ermA (45.54%) and ereA (40.59%), while, ermB was present with a low detection rate (11.88%). Other drug-resistant genes test results showed that fexA, tetL, aadA1, aph (4') - Ia detection rate was higher, which was 92.10%, 97.02%, 97.29% and 83.17%, respectively; secondly, tet (M), the detection rate was 71.29%; the detection rate of aac (6') - Ib, aac (3') - Ic, aph (3') -II, aph (3') -IV, optrA, tet (A) and tet (C) was relatively low, which was 49.50%, 46.53%, 39.60%, 37.62%, 33.67%, 29.70% and 20.79%, respectively; the detection rate of cfr, tet (K), lnuA and lnuF was the lowest, which was 17.82%, 14.85%, 5.94% and 3.96%, respectively. The relationship of the number of MLSB resistance genes and the number of resistance to antibiotics was further analyzed, which indicated the increased number of resistance to antibiotics was corresponding with the increased number of MLSB resistance genes and high detection rate of other resistant genes among the LA-MRSA isolates. 【Conclusion】In conclusion, in this study, a high resistance to MLSB was found and it was consistent to the high detection rate of ermC gene among LA-MRSA isolates. The multi-drug resistance of LA-MRSA isolates was serious and this was closely related to the number of MLSB resistance genes. Therefore, it is urgently needed to take action in the rational use of antibiotics, as to reducing the emergence of multi-drug resistance LA-MRSA.

Key words: food animal, LA-MRSA, MLSB, erm, multidrug resistance

Table 1

Primer sequences of the detection of MLSB resistant genes"

基因名称Genes’ name 引物序列（5′→3′）Primer sequence 片段长度Size (bp) t退火 (℃) 参考文献Reference
mecA F: AAAATCGATGGTAAAGGTTGGC
R: AGTTCTGCAGTACCGGATTTGC		 533 58 [22]
mecC F:4GAAAAAAAGGCTTAGAACGCCTC
R:GAAGATCTTTTCCGTTTTCAGC		 137 58 [23]
ermA F: GTTCAAGAACAATCAATACAGAG 421 52 [24]
R: GGATCAGGAAAAGGACATTTTAC
ermB F:CCGTTTACGAAATTGGAACAGGTAAAGGGC 359 55 [24]
R: GAATCGAGAC TTGAGTGTGC
ermC F: GCTAATATTG TTTAAATCGT CAATTCC 572 52 [25]
R: GGATCAGGAA AAGGACATTT TAC
ereA F: AACACCCTGAACCCAAGGGACG 420 50 [26]
R: CTTCACATCCGGATTCGCTCGA
ereB F: AGAAATGGAGGTTCATACTTACCA 546 50 [26]
R: CATATAATCATCACCAATGGCA

Table 2

Isolation of LA-MRSA"

样品来源
Source		 样品数
Number		 金黄色葡萄球菌数(株)
Number of Staphylococcus aureus		 MRSA菌株数(株)
Number of MRSA		 样品中MRSA菌株 的分离率
The separation rate of MRSA (%)		 金黄色葡萄球菌中 MRSA菌的检出率
The separation rate of Staphylococcus aureus (%)
猪源Pig 粪便Faeces 3020 177 40 1.32% 22.59%
鼻拭子Nose swab 480 104 36 7.50% 34.60%
鸡源Chicken 盲肠Cecum 1546 109 14 0.91% 12.84%
肝脏Liver 654 48 6 0.92% 12.50%
鸭源Duck 盲肠Cecum 436 24 3 0.69% 12.50%
肝脏Liver 364 18 2 0.55 11.1%
总数Total 6500 480 101 1.55% 21.04%

Table 3

The result of multiple drug resistance of MRSA from animals"

对抗菌药物耐药的数目 Number 2 6 7 9 10 11 12
菌株数（株）The number of strain 1 1 2 3 35 48 11
百分率Percentage (%) 0.98 0.98 1.98 3.96 34.65 47.52 10.89

Fig. 1

Resistant rates to 14 antibiotics in 101 LA-MRSA strains"

Fig. 2

The detection rates of other drug resistance genes among MLSB resistant LA-MRSA"

Table 4

The relationship between genetic combination of MLSB and multiple drug resistance of LA-MRSA"

多重耐药（菌株数）
Multi-drug (Number of strain)		 基因组合
Assortment of genes		 菌株数
The number of strain		 携带相同个数耐药基因所占比例
Percentage (%)
12耐（11） ermC-ereB-ermA-ermB-ereA 1 1（9.09%）
ermC-ereB-ermA-ermB 2 5（45.45%）
ermC-ereB-ermA-ereA 2
ermC-ereB-ermB-ereA 1
ermC-ereB-ermA 4 4（36.36%）
ermC-ereB 1 1（9.09%）
11耐（48） ermC-ereB-ermA-ereA 11 11（22.92%）
ermC-ereB-ereA 6 17（35.42%）
ermC-ereB-ermA 9
ermC-ereB-ermB 1
ermC-ereA-ermA 1
ermC-ereB 11 19（39.58%）
ermC-ermA 5
ermC-ereA 3
ermC 2 2（4.17%）
10耐（35） ermC-ereB-ermA-ereA 3 7（20.00%）
ermC-ereB-ermA-ermB 4
ermC-ermB-ermA 2 15（42.86%）
ermC-ereB-ereA 4
ermC-ereB-ermA 8
ermC-ereA-ermA 1
ermC-ereB 7 9（25.71%）
ermC-ermB 2
ermC 4 4（11.43%）
9耐（3） ermC-ereB-ereA 1 1(33.33%)
ermC-ereA 1 2(66.67%)
ermC-ereB 1
7耐（2） ermC-ereB-ermA-ermB 1 1(50.00%)
ermC-ermA 1 1(50.00%)
6耐（1） ermC-ereB 1 1(100.00%)
2耐（1） ermC-ereB-ereA 1 1(100.00%)
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