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Genomic characterisation, detection of genes encoding virulence factors and evaluation of antibiotic resistance of Trueperella pyogenes isolated from cattle with clinical metritis

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Abstract

Trueperella pyogenes is one of the most important microorganisms causing metritis in post-partum cattle. Co-infection with other bacterial species such as Escherichia coli or Fusobacterium necrofurom increases the severity of the disease and the persistence of bacteria in utero. The aim of this study was to investigate the frequency of T. pyogenes strains, and their virulence and antimicrobial resistant profiles in metritis cases. The study was carried out on 200 samples obtained from metritis discharges of postpartum cattle on 18 farms around Tehran, Iran. Sixty-five T. pyogenes isolates (32.5%) were identified, of which 16 isolates were detected as pure cultures and the other 49 isolates from cultures most commonly mixed with E. coli or F. necrofurom. In terms of diversity in biochemical characteristic of T. pyogenes strains, 8 different biotypes were identified among the isolates. Single or multi antimicrobial resistance was observed in 48 isolates (73.9%), which was mostly against trimethoprim sulfamethoxazole, azithromycin, erythromycin and streptomycin. The tetracycline resistance gene tetW and macrolide resistance genes ermB and ermX were detected in 30, 18 and 25 isolates, respectively. In the screening of genes encoding virulence factors, fimA and plo genes were identified in all tested isolates. Genes encoding nanP, nanH, fimC, fimG, fimE and cbpA were detected in 50, 54, 45, 40, 50 and 37 of isolates, respectively. Thirteen different genotypes were observed in these T. pyogenes isolates. A significant association between clonal types and virulence factor genes, biochemical profile, CAMP test result, severity of the disease and sampling time was detected.

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Abbreviations

T.:

Trueperella

Plo :

Pyolysin

nanH :

Neuraminidase H

nanP :

Neuraminidase P

fimA :

Type A fimbria

fimC :

Type C fimbria

fimG :

Type G fimbria

fimE :

Type E fimbria

cbpA :

Collagen-binding protein A

tetW :

Tetracycline W

ermX :

Erythromycin X

bp:

Base pair/s

µl:

Microliter

°C:

Centigrade

V:

Voltage

µm:

Micromol

TSB:

Tryptic soy broth

ISR:

Intergenic spacer region

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Acknowledgements

This project emanates from Ph.D. thesis of the first author and was financially supported by research grants from Bu-Ali Sina University of Hamedan. The authors would like to thank Mottaghian P. for help in sample collection.

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Authors and Affiliations

  1. Department of Pathobiology, Faculty of Veterinary Science, Bu-Ali Sina University, Hamedan, Iran

    Iradj Ashrafi Tamai, Abdolmajid Mohammadzadeh & Pezhman Mahmoodi

  2. Department of Microbiology and Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

    Taghi Zahraei Salehi

Authors
  1. Iradj Ashrafi Tamai
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  2. Abdolmajid Mohammadzadeh
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  3. Taghi Zahraei Salehi
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  4. Pezhman Mahmoodi
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These authors have contributed equally to this work.

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Correspondence to Abdolmajid Mohammadzadeh.

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The authors declare that they have no conflict of interest.

Ethical approval

We verify that the current study has been carried out under ethical regulations. All animal experiments including the sampling procedure were done considering animal welfare and relief. Furthermore, no harmful techniques or procedures to animal health were used in this study.

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Ashrafi Tamai, I., Mohammadzadeh, A., Zahraei Salehi, T. et al. Genomic characterisation, detection of genes encoding virulence factors and evaluation of antibiotic resistance of Trueperella pyogenes isolated from cattle with clinical metritis. Antonie van Leeuwenhoek 111, 2441–2453 (2018). https://doi.org/10.1007/s10482-018-1133-6

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