Stiftung Tierärztliche Hochschule Hannover (TiHo)

Plasmid-located dfrA14 gene in Pasteurella multocida isolates from three different pig-producing farms in Germany

Affiliation
Institute of Microbiology and Epizootics, Freie Universität Berlin, Berlin, Germany; Institute of Farm Animal Genetics, Friedrich-Loeffler-Institut (FLI), Neustadt-Mariensee, Germany.
Niemann, Lisa;
Affiliation
Institute of Microbiology and Epizootics, Freie Universität Berlin, Berlin, Germany.
Feudi, Claudia;
Affiliation
Institute of Microbiology and Epizootics, Freie Universität Berlin, Berlin, Germany.
Eichhorn, Inga;
Affiliation
Institute of Microbiology and Epizootics, Freie Universität Berlin, Berlin, Germany.
Hanke, Dennis;
Affiliation
Department of Pharmacology, Toxicology and Pharmacy, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany.
Müller, Petra;
Affiliation
Clinic for Swine and Small Ruminants and forensic Medicine and Ambulatory Services, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany.
Brauns, Jasmin;
Affiliation
Vet-Team Reken, Reken, Germany.
Nathaus, Rolf;
GND
1207174033
Affiliation
Institute for Biometry, Epidemiology and Information Processing, WHO Collaborating Centre for Research and Training for Health at the Human-Animal-Environment Interface, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany.
Schäkel, Franziska;
GND
138971609
ORCID
0000-0002-2738-4257
Affiliation
Clinic for Swine and Small Ruminants and forensic Medicine and Ambulatory Services, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany.
Höltig, Doris;
ORCID
0000-0002-3545-0859
Affiliation
Clinic for Swine and Small Ruminants and forensic Medicine and Ambulatory Services, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany.
Wendt, Michael;
Affiliation
Institute of Farm Animal Genetics, Friedrich-Loeffler-Institut (FLI), Neustadt-Mariensee, Germany.
Kadlec, Kristina;
Affiliation
Institute of Microbiology and Epizootics, Freie Universität Berlin, Berlin, Germany. Electronic address: stefan.schwarz@fu-berlin.de.
Schwarz, Stefan

Pasteurella multocida is an important respiratory tract pathogen in intensive livestock farming, especially in pigs. Antimicrobial agents are frequently used to combat infections caused by this pathogen. In a study on antimicrobial resistance among respiratory tract pathogens of pigs from 30 German pig-producing farms, P. multocida isolates (n = 9) with high minimal inhibitory concentration (MIC) values of 16/304 mg/L (n = 2), 32/608 mg/L (n = 3) or ≥64/1216 mg/L (n = 4) for trimethoprim/sulfamethoxazole (1:19) and of ≥512 mg/L (n = 9) for trimethoprim (TMP) were detected in three of these farms. The genetic relatedness of the isolates was investigated via capsule-specific PCR and macrorestriction analyses with ApaI and SmaI. Pulsed-field gel electrophoresis revealed indistinguishable restriction patterns per farm, with slight differences between the three farms. All isolates represented capsular type A. Four representative isolates, that were subjected to whole genome sequencing, shared the multi-locus sequence type (ST) 3. Their plasmids were transformed into E. coli TOP10 with subsequent selection on TMP-containing agar plates. Antimicrobial susceptibility testing and plasmid analysis of the transformants confirmed that they were resistant to sulfonamides and trimethoprim and carried only a single small plasmid. This plasmid was completely sequenced and revealed a size of 6050 bp. Sequence analyses identified the presence of a resistance gene cluster comprising the genes sul2-ΔstrA-dfrA14-ΔstrA-ΔstrB. Further analysis identified a dfrA14 gene cassette being integrated into the strA reading frame. Neither the gene dfrA14 nor this gene cluster have been detected before in P. multocida.

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