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Acheamfour, Chanelle L; Parveen, Salina; Gutierrez, Alan; Handy, Eric T; Behal, Sara; Kim, Donghyun; Kim, Seongyun; East, Cheryl; Xiong, Ray; Haymaker, Joseph R; et al(
, Microbiology Spectrum)
Wang, Luxin
(Ed.)
ABSTRACT
Alternative irrigation waters (rivers, ponds, and reclaimed water) can harbor bacterial foodborne pathogens likeSalmonella entericaandListeria monocytogenes, potentially contaminating fruit and vegetable commodities. Detecting foodborne pathogens using qPCR-based methods may accelerate testing methods and procedures compared to culture-based methods. This study compared detectionof S. enterica and L. monocytogenesby qPCR (real-time PCR) and culture methods in irrigation waters to determine the influence of water type (river, pond, and reclaimed water), season (winter, spring, summer, and fall), or volume (0.1, 1, and 10 L) on sensitivity, accuracy, specificity, and positive (PPV), and negative (NPV) predictive values of these methods. Water samples were collected by filtration through modified Moore swabs (MMS) over a 2-year period at 11 sites in the Mid-Atlantic U.S. on a bi-weekly or monthly schedule. For qPCR, bacterial DNA from culture-enriched samples (n= 1,990) was analyzed by multiplex qPCR specific forS. entericaandL. monocytogenes. For culture detection, enriched samples were selectively enriched, isolated, and PCR confirmed. PPVs for qPCR detection ofS. entericaandL. monocytogeneswere 68% and 67%, respectively. The NPV were 87% (S. enterica) and 85% (L. monocytogenes). Higher levels of qPCR/culture agreement were observed in spring and summer compared to fall and winter forS. enterica; forL. monocytogenes, lower levels of agreement were observed in winter compared to spring, summer, and fall. Reclaimed and pond water supported higher levels of qPCR/culture agreement compared to river water for bothS. entericaandL. monocytogenes, indicating that water type may influence the agreement of these results.
IMPORTANCE
Detecting foodborne pathogens in irrigation water can inform interventions and management strategies to reduce risk of contamination and illness associated with fresh and fresh-cut fruits and vegetables. The use of non-culture methods like qPCR has the potential to accelerate the testing process. Results indicated that pond and reclaimed water showed higher levels of agreement between culture and qPCR methods than river water, perhaps due to specific physiochemical characteristics of the water. These findings also show that season and sample volume affect the agreement of qPCR and culture results. Overall, qPCR methods could be more confidently utilized to determine the absence ofSalmonella entericaandListeria monocytogenesin irrigation water samples examined in this study.
Improving the microbial quality of agricultural water through filtration can benefit small farms globally. The incorporation of zero-valent iron (ZVI) into sand filters (ZVI–sand) has been effective in reducing E. coli, Listeria spp., and viruses from agricultural water. This study evaluated ZVI–sand filtration in reducing E. coli levels based on influent water type and the percentage of ZVI in sand filters. A ZVI–sand filter (50% ZVI/50% sand) significantly (p < 0.001) reduced E. coli levels in deionized water by more than 1.5 log CFU/mL compared to pond water over six separate trials, indicating that water type impacts E. coli removal. Overall reductions in E. coli in deionized water and pond water were 98.8 ± 1.7% and 63 ± 24.0% (mean ± standard deviation), respectively. Filters constructed from 50% ZVI/50% sand showed slightly more reduction in E. coli in pond water than filters made from a composition of 35% ZVI/65% sand; however, the difference was not statistically significant (p = 0.48). Principal component analysis identified that the turbidity and conductivity of influent water affected E. coli reductions in filtered water in this study. ZVI–sand filtration reduces Escherichia coli levels more effectively in waters that contain low turbidity values.
Acheamfour, Chanelle L.; Parveen, Salina; Hashem, Fawzy; Sharma, Manan; Gerdes, Megan E.; May, Eric B.; Rogers, Koriante; Haymaker, Joseph; Duncan, Rico; Foust, Derek; et al(
, Microbiology Spectrum)
Gralnick, Jeffrey A.
(Ed.)
ABSTRACT Irrigation water sources have been shown to harbor foodborne pathogens and could contribute to the outbreak of foodborne illness related to consumption of contaminated produce. Determining the probability of and the degree to which these irrigation water sources contain these pathogens is paramount. The purpose of this study was to determine the prevalence of Salmonella enterica and Listeria monocytogenes in alternative irrigation water sources. Water samples ( n = 188) were collected over 2 years (2016 to 2018) from 2 reclaimed water plants, 3 nontidal freshwater rivers, and 1 tidal brackish river on Maryland’s Eastern Shore (ESM). Samples were collected by filtration using modified Moore swabs (MMS) and analyzed by culture methods. Pathogen levels were quantified using a modified most probable number (MPN) procedure with three different volumes (10 liters, 1 liter, and 0.1 liter). Overall, 65% (122/188) and 40% (76/188) of water samples were positive for S. enterica and L. monocytogenes , respectively. For both pathogens, MPN values ranged from 0.015 to 11 MPN/liter. Pathogen levels (MPN/liter) were significantly ( P < 0.05) greater for the nontidal freshwater river sites and the tidal brackish river site than the reclaimed water sites. L. monocytogenes levels in water varied based on season. Detection of S. enterica was more likely with 10-liter filtration compared to 0.1-liter filtration. The physicochemical factors measured attributed only 6.4% of the constrained variance to the levels of both pathogens. This study shows clear variations in S. enterica and L. monocytogenes levels in irrigation water sources on ESM. IMPORTANCE In the last several decades, Maryland’s Eastern Shore has seen significant declines in groundwater levels. While this area is not currently experiencing drought conditions or water scarcity, this research represents a proactive approach. Efforts, to investigate the levels of pathogenic bacteria and the microbial quality of alternative irrigation water are important for sustainable irrigation practices into the future. This research will be used to determine the suitability of alternative irrigation water sources for use in fresh produce irrigation to conserve groundwater.
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