Characterization and long-read sequencing of biofilms formed by the microbiota present on inert surfaces in poultry slaughterhouses

Cross-contamination from inert slaughterhouse surfaces is among the main sources of contamination of poultry. The objective of the research reported here was to characterize the biofilms formed by the microbiota present on various surfaces in two poultry slaughterhouses in north-western Spain. Forty-four samples (22 from each slaughterhouse) were taken by swab rubbing at different points along the processing line (from stunning to cutting). The microbiota on all surfaces was able to form biofilms, which were studied by scanning confocal laser microscopy. The total biovolume in the observation field of 16,078.24 μm² ranged from 22,106.8 ± 5544.3 μm³ to 414,229.6 ± 1621.0 μm³. Average values were higher in abattoir A than in abattoir B, with significant differences (P < 0.05) between surfaces. The percentage of biovolume of Gram-positive bacteria ranged between 0.02 % and 5.38 %. The highest percentages of Gram-positive bacteria were detected towards the beginning of the processing line. The microbiota of the biofilms was identified using long-read sequencing techniques (Oxford Nanopore). The predominant genera (found in >50.0 % of the biofilms) were Pseudomonas, Citrobacter, Klebsiella, Serratia, Escherichia, Enterobacter, Stenotrophomonas, Salmonella, Shewanella, Acinetobacter and Aeromonas. In addition, some pathogenic bacteria were detected, including Salmonella (31 surfaces), Yersinia enterocolitica (12), Escherichia coli O157:H7 (6), Campylobacter spp. (4) and Listeria monocytogenes (3). This research work has permitted identification of the most contaminated surfaces in poultry abattoirs and can serve as a starting point for the design of more effective cleaning and disinfection protocols.