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Streptococcus suis  ( S. suis ) and  Actinobacillus pleuropneumoniae  ( A. pleuropneumoniae ) are primary swine pathogens that have been frequently co-isolated from pigs suffering from severe respiratory disease. The purpose of this study was to investigate the biological impacts of the interactions between  S. suis  and  A. pleuropneumoniae . A single- and dual-species culture model was established  in vitro  via  S. suis  HA9801 (serotype 2) and  A. pleuropneumoniae  CVCC265 (serotype 1). The single or mixed biofilms were imaged by confocal laser scanning microscopy. The biomass and viable cells in biofilms were quantified by crystal violet staining and determination of colony-forming units. The antibiotic susceptibility was determined by a microdilution broth method. The differences in gene transcription in pure- or mixed-species biofilms of  S. suis  and  A. pleuropneumoniae  was evaluated by quantitative PCR.  S. suis  and  A. pleuropneumoniae  formed two-species biofilms when co-cultured  in vitro . When co-cultured with  S. suis , biofilm formation by  A. pleuropneumoniae  was significantly increased with the absence of NAD that is necessary for the growth of  A. pleuropneumoniae . Moreover, compared with monocultures, the antibiotic resistance of  S. suis  and  A. pleuropneumoniae  was both enhanced in the co-culture model. When grown in dual-species biofilms, for  A. pleuropneumoniae , genes associated with virulence factors, including exotoxins and adhesins, were significantly upregulated. For  S. suis , virulence factor-related genes  cps2 ,  gdh ,  mrp , and  sly  were highly induced. These results suggest that the interspecies interactions between  S. suis  and  A. pleuropneumoniae  may be cooperative under specific conditions and may play an important role in the disease progression and persistent infection.

frontiers in microbiology

In vitro Mixed Biofilm of Streptococcus suis and Actinobacillus pleuropneumoniae Impacts Antibiotic Susceptibility and Modulates Virulence Factor Gene Expression