Large genetic diversity of Arcobacter butzleri isolated from raw milk in Southern Italy

Abstract

Arcobacter butzleri is a zoonotic foodborne pathogen able to cause enteric and extraintestinal diseases. Its occurrence in foodstuff is well recognized worldwide but data on its presence in foods from Southern Italy are scarce. In this study the results on the occurrence and genotyping of Arcobacter spp. in bulk milk samples collected in Southern Italy are reported. Out of 484 samples, 64 (13.2%) resulted positive for the presence of Arcobacter spp. Using Real Time PCR but as few as 31.2% of these samples turned out as positive by using the cultural method, showing an overall prevalence of 4.1%. All isolates were identified as A. cryaerophilus using the biochemical identification whilst the sequencing of the atpA gene revealed that all the isolates were A. butzleri. Among the confirmed isolates, 16 different Sequence Types (ST) were identified using the Multi Locus Sequence Typing (MLST), 14 (87.5%) of which were previously unreported. Our survey reveals the presence of A. butzleri in bulk tank milk from Southern Italy and highlights the discrepancy between the two approaches used both for the detection (i.e., real time PCR vs cultural method) and the identification (i.e., biochemical test vs aptA sequencing) of Arcobacter spp In addition, a large genetic diversity among the isolates was detected and this makes the identification of source of the infections very challenging in outbreaks investigation.

Keywords: Arcobacter; Bulk tank milk; Genotyping; Multi locus sequence typing (MLST); Real-time PCR.