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Chlamydia suis  is a swine pathogen that has also recently been found to cause zoonotic infections of the human eye, pharynx, and gastrointestinal tract. Many strains contain a tetracycline class C gene [ tet (C)] cassette that confers tetracycline resistance. The cassette was likely originally acquired by horizontal gene transfer from a Gram-negative donor after the introduction of tetracycline into animal feed in the 1950s. Various research groups have described the capacity for different  Chlamydia  species to exchange DNA by homologous recombination. Since over 90% of  C. suis  strains are tetracycline resistant, they represent a potential source for antibiotic-resistance spread within and between  Chlamydiaceae  species. Here, we examined the genetics of  tet (C)-transfer among  C. suis  strains. Tetracycline-sensitive  C. suis  strain S45 was simultaneously or sequentially co-infected with tetracycline-resistant  C. suis  strains in McCoy cells. Potential recombinants were clonally purified by a harvest assay derived from the classic plaque assay.  C. suis  strain Rogers132, lacking transposases IS 200  and IS 605 , was the most efficient donor, producing two unique recombinants detected in three of the 56 (5.4%) clones screened. Recombinants were found to have a minimal inhibitory concentration (MIC) of 8-16 μg/mL for tetracycline. Resistance remained stable over 10 passages as long as recombinants were initially grown in tetracycline at twice the MIC of S45 (0.032 μg/mL). Genomic analysis revealed that  tet (C) had integrated into the S45 genome by homologous recombination at two unique sites depending on the recombinant: a 55 kb exchange between  nrq F and  pck G, and a 175 kb exchange between  kds A and  cys Q. Neither site was associated with inverted repeats or motifs associated with recombination hotspots. Our findings show that cassette transfer into S45 has low frequency, does not require IS 200 /IS 605  transposases, is stable if initially grown in tetracycline, and results in multiple genomic configurations. We provide a model for stable cassette transfer to better understand the capability for cassette acquisition by  Chlamydiaceae  species that infect humans, a matter of public health importance.

frontiers in microbiology

Tet(C) Gene Transfer between Chlamydia suis Strains Occurs by Homologous Recombination after Co-infection: Implications for Spread of Tetracycline-Resistance among Chlamydiaceae