Variability in cadmium tolerance of closely related Listeria monocytogenes isolates originating from dairy processing environments

Abstract

Increased tolerance to cadmium in Listeria monocytogenes has been suggested to contribute to their persistence in natural and food production environments. This study investigated the phenotypic cadmium response of L. monocytogenes strains with efflux pump cadAC (variants 1–4) and related strains with cadA1C1 . Growth of cadAC variant strains ( n = 5) in 0 µM–120 µM cadmium salts (CdCl 2 , CdSO 4 ) in Mueller-Hinton broth (MHB) was evaluated. Additionally, 88 L . monocytogenes strains from dairy processing facilities were exposed to 43.8 µM CdCl 2 in MHB, and their lag phase duration (LPD) was measured. Strains with cadA1 through cadA3 showed similar growth trends in the presence of cadmium, while the cadA4 variant (Scott A) had the highest CdCl 2 minimum inhibitory concentration (175 µM). Growth varied between the two salts, with CdSO 4 significantly increasing LPD ( P < 0.05) compared to CdCl 2 . In 43.8 µM CdCl 2 , cadA1 strains displayed LPDs ranging from 0.99 ± 0.14 h to 6.44 ± 0.08 h, with no clear genomic differences explaining this variability. Strains without cadA did not grow at 43.8 µM CdCl 2 but exhibited low tolerance (10.9 µM CdCl 2 ), potentially due to non-specific soft metal ATPases (626 aa; 737 aa) and soft metal resistance proteins encoded by czc genes (289 aa; 291 aa; 303 aa) within their chromosomes. These findings enhance our understanding of L. monocytogenes cadmium tolerance and underscore the need for further research to explore the genetic and physiological factors underlying these trends. IMPORTANCE Mobile genetic elements in Listeria monocytogenes contribute to its survival in natural and food processing environments. This study focused on how different genetic variants of the efflux pump gene cadAC and group of closely related cadA1C1 strains respond to cadmium exposure. When exposed to two cadmium salts, cadmium chloride and cadmium sulfate, we observed varying growth patterns, with a significantly longer lag phase in cadmium sulfate compared to cadmium chloride. Strains with cadA1 to cadA3 had similar growth trends, whereas a strain with the cadA4 variant had the highest minimum inhibitory concentration value. Among 88 strains from dairy processing facilities, significant phenotypic differences were observed despite core genome similarities, indicating other underlying genetic and physiological factors contribute to cadmium tolerance. Since cadmium tolerance studies in L. monocytogenes are limited, with rare phenotypic comparisons between closely related strains, our study makes an important observation and contribution to understanding of L. monocytogenes tolerance to cadmium by providing phenotypic comparisons between numerous strains within the same clonal group (<16 single nucleotide polymorphisms).