In September 2018, Hurricane Florence caused extreme flooding in eastern North Carolina, USA, a region highly dense in concentrated animal production, especially swine and poultry. In this study, floodwater samples (n=96) were collected as promptly post-hurricane as possible and for up to approx. 30 days, and selectively enriched for Campylobacter using Bolton broth enrichment and isolation on mCCDA microaerobically at 42°C. Only one sample yielded Campylobacter, which was found to be Campylobacter jejuni with the novel genotype ST-2866. However, the methods employed to isolate Campylobacter readily yielded Arcobacter from 73.5% of the floodwater samples. The Arcobacter isolates failed to grow on Mueller-Hinton agar at 25, 30, 37 or 42°C microaerobically or aerobically, but could be readily subcultured on mCCDA at 42°C microaerobically. Multilocus sequence typing of 112 isolates indicated that all were Arcobacter butzleri. The majority (85.7%) of the isolates exhibited novel sequence types (STs), with 66 novel STs identified. Several STs, including certain novel ones, were detected in diverse waterbody types (channel, isolated ephemeral pools, floodplain) and from multiple watersheds, suggesting the potential for regionally-dominant strains. The genotypes were clearly partitioned into two major clades, one with high representation of human and ruminant isolates and another with an abundance of swine and poultry isolates. Surveillance of environmental waters and food animal production systems in this animal agriculture-dense region is needed to assess potential regional prevalence and temporal stability of the observed A. butzleri strains, as well as their potential association with specific types of food animal production. IMPORTANCE Climate change and associated extreme weather events can have massive impacts on the prevalence of microbial pathogens in floodwaters. However, limited data are available on foodborne zoonotic pathogens such as Campylobacter or Arcobacter in hurricane-associated floodwaters in rural regions with intensive animal production. With high density of intensive animal production as well as pronounced vulnerability to hurricanes, Eastern North Carolina presents unique opportunities in this regard. Our findings revealed widespread incidence of the emerging zoonotic pathogen Arcobacter butzleri in floodwaters from Hurricane Florence. We encountered high and largely unexplored diversity while also noting the potential for regionally-abundant and persistent clones. We noted pronounced partitioning of the floodwater genotypes in two source-associated clades. The data will contribute to elucidating the poorly-understood ecology of this emerging pathogen, and highlight the importance of surveillance of floodwaters associated with hurricanes and other extreme weather events for Arcobacter and other zoonotic pathogens.

Multidrug resistance (MDR) in foodborne pathogens is a major food safety and public health issue. Here we describe whole-genome sequences of two MDR strains of Campylobacter jejuni and Campylobacter coli from turkey feces and a housefly from a turkey farm. Both strains harbor a novel chromosomal gentamicin resistance mobile element.

is a leading foodborne pathogen, and poultry products are major vehicles for human disease. However, determinants impacting colonization in poultry remain poorly understood, especially with turkeys. Here, we used a paired-farm design to concurrently investigate colonization and strain types in two turkey breeds (Hybrid and Nicholas) at two farms in eastern North Carolina. One farm (the Teaching Animal Unit [TAU]) was a university teaching unit at least 40 km from commercial turkey farms, while the other (SIB) was a commercial farm in an area with a high density of turkey farms. Day-old birds were obtained from the same breeder flock and hatchery and placed at TAU and SIB on the same day. Birds were marked to identify turkey breed and then commingled on each farm. TAU birds became colonized 1 week later than SIB and had lower initial levels in the cecum. Interestingly, genotypes and antimicrobial resistance profiles differed markedly between the farms. Most TAU isolates were resistant only to tetracycline, whereas multidrug-resistant isolates predominated at SIB. Multilocus sequence typing revealed that no genotypes were shared between TAU and SIB. A bovine-associated genotype (sequence type 1068 [ST1068]) predominated in from TAU, while SIB isolates had genotypes commonly encountered in commercial turkey production in the region. One multidrug-resistant strain (ST1839) showed significant association with one of the two turkey breeds. The findings highlight the need to further characterize the impact of farm-specific factors and host genetics on antimicrobial resistance and genotypes of and that colonize turkeys. Colonization of poultry with at the farm level is complex, poorly understood, and critically linked to contamination of poultry products, which is known to constitute a leading risk factor for human campylobacteriosis. Here, we investigated the use of a paired-farm design under standard production conditions and in the absence of experimental inoculations to assess potential impacts of farm and host genetics on prevalence, antimicrobial resistance and genotypes of in commercial turkeys of two different breeds. Data suggest impacts of farm proximity to other commercial turkey farms on the onset of colonization, genotypes, and antimicrobial resistance profiles of colonizing the birds. Furthermore, the significant association of a specific multidrug-resistant strain with turkeys of one breed suggests colonization partnerships at the strain-turkey breed level. The study design avoids potential pitfalls associated with experimental inoculations, providing novel insights into the dynamics of turkey colonization with in actual farm ecosystems.

Listeria monocytogenes has been extensively studied as a model facultative intracellular pathogen. While the roles of major virulence factors in host-pathogen interactions have been extensively characterized, recent work suggests that some of these factors can also contribute to environmental proliferation of this pathogen. In this study, we characterized two non-hemolytic transposon mutants of strain 2011L-2858 (serotype 1/2b), implicated in the 2011 listeriosis outbreak via whole cantaloupe, for their capacity to form biofilms on polystyrene, aggregate, and colonize cantaloupe rind. One mutant harbored a single mariner-based transposon insertion in hly, encoding the hemolysin Listeriolysin O, while the other harbored a single insertion in prfA, encoding PrfA, a master regulator for hly and numerous other virulence genes. Biofilm formation was significantly reduced in the prfA mutant, and to a lesser extent, in the hly mutant. Inactivation of either hly or prfA significantly reduced L. monocytogenes aggregation. However, both mutants adhered similarly to the wildtype parental strain on cantaloupe rind at either 25 or 37°C. Furthermore, growth and competitive fitness of the mutants on cantaloupe rind was not significantly impacted at either temperature. The findings suggest that, in spite of their involvement in biofilm formation and aggregation, these key virulence determinants may not be required for the ability of L. monocytogenes to colonize fresh produce.

Hurricane Florence brought unprecedented rainfall and flooding to Eastern North Carolina in 2018. Extensive flooding had the potential to mobilize microbial contaminants from a variety of sources. Our study evaluated microbial contaminants in surface waters at 40 sites across Eastern North Carolina 1 week after the hurricane made landfall (Phase 1) and one month later (Phase 2). High concentrations of Escherichia coli were detected in flowing channel and floodwater samples across both phases; however, channel samples during Phase 2 had higher concentrations of E. coli compared to Phase 1. Human- and swine-associated fecal markers were detected in 26% and 9% of samples, respectively, with no trends related to phase of sampling. Arcobacter butzleri was previously shown to be recovered from most (73%) samples, and detection of this pathogen was not associated with any source-associated fecal marker. Detection of Listeria spp. was associated with the swine-associated fecal marker. These results suggest that improved swine and human feces management should be explored to prevent microbial contamination in surface water, especially in regions where extreme rainfall may increase due to climate change. Sampling at higher frequency surrounding rainfall events would provide more detailed characterization of the risks posed by floodwater at different time scales and under different antecedent conditions.