Fecal Contamination in the Surface Waters of a Rural- and an Urban-Source Watershed.

Surface waters are commonly used as source water for drinking water and irrigation. Knowledge of sources of fecal pollution in source watersheds benefits the design of effective source water protection plans. This study analyzed the relationships between enteric pathogens ( O157:H7, spp., and spp. [, and ]), water quality (turbidity, temperature, and ), and human and ruminant-cow and mitochondrial DNA (mtDNA)-based fecal source tracking (FST) markers in two source watersheds. Water samples ( = 329) were collected at 10 sites (five in each watershed) over 18 mo. The human marker (HF183) occurred in 9 to 10% of the water samples at nine sampling sites; while a forested site in the urban watershed tested negative. Ruminant-cow markers (BacR and CowM2) only appeared in the rural watershed (6%). The mtDNA markers (HcytB and AcytB) showed the same pattern but were less sensitive due to lower fecal concentrations. Higher prevalences ( < 0.05) of spp. (41 vs. 16% for the rural and urban watershed, respectively) and O157:H7 (12 vs. 3%) were observed in the rural watershed, while spp. levels were comparable (23-28%). Densities of ≥100 colony-forming units (CFU) 100 mL increased the odds ( < 0.05) of detecting the enteric bacterial pathogens. The water turbidity levels (nephelometric turbidity units [NTU] ≥ 1.0) similarly predicted ( < 0.05) pathogen presence. Storm events increased ( < 0.01) pathogen and fecal marker concentrations in the waterways. The employment of multiple FST methods suggested failing onsite wastewater systems contribute to human fecal pollution in both watersheds.

Comparison of the Prevalences and Diversities of Listeria Species and Listeria monocytogenes in an Urban and a Rural Agricultural Watershed.

Foods and related processing environments are commonly contaminated with the pathogenic Listeria monocytogenes. To investigate potential environmental reservoirs of Listeria spp. and L. monocytogenes, surface water and point source pollution samples from an urban and a rural municipal water supply watershed in Nova Scotia, Canada, were examined over 18 months. Presumptive Listeria spp. were cultured from 72 and 35% of rural and urban water samples, respectively, with 24% of the positive samples containing two or three different Listeria spp. The L. innocua (56%) and L. welshimeri (43%) groups were predominant in the rural and urban watersheds, respectively. Analysis by the TaqMan assay showed a significantly (P < 0.05) higher prevalence of L. monocytogenes of 62% versus 17% by the culture-based method. Both methods revealed higher prevalences in the rural watershed and during the fall and winter seasons. Elevated Escherichia coli (≥ 100 CFU/100 ml) levels were not associated with the pathogen regardless of the detection method. Isolation of Listeria spp. were associated with 70 times higher odds of isolating L. monocytogenes (odds ratio = 70; P < 0.001). Serogroup IIa was predominant (67.7%) among the 285 L. monocytogenes isolates, followed by IVb (16.1%), IIb (15.8%), and IIc (0.4%). L. monocytogenes was detected in cow feces and raw sewage but not in septic tank samples. Pulsotyping of representative water (n = 54) and local human (n = 19) isolates suggested genetic similarities among some environmental and human L. monocytogenes isolates. In conclusion, temperate surface waters contain a diverse Listeria species population and could be a potential reservoir for L. monocytogenes, especially in rural agricultural watersheds.

Role of initial contamination levels, biofilm maturity and presence of salt and fat on desiccation survival of Listeria monocytogenes on stainless steel surfaces.

This study investigated the effect of initial contamination levels, biofilm maturity and presence of salt and fatty food soils on desiccation survival of Listeria monocytogenes on stainless steel (SS) coupons. L. monocytogenes cultures grown (at 15 °C for 48 h) in Tryptic Soy Broth with 1% glucose (TSB-glu) containing either 0.5 or 5% (w/v) NaCl were re-suspended in TSB-glu containing either 0.5 or 5% NaCl and used to contaminate SS coupons at levels of 3.5, 5.5, and 7.5 log CFU/cm². Desiccation (at 15 °C for 20 days, 43% RH) commenced immediately (non-biofilm) or following biofilm formation (at 15 °C for 48 h, 100% RH). To study the impact of food lipids, non-biofilm L. monocytogenes cells were suspended in TSB-glu containing either canola oil (5-10%) or lard (20-60%) and desiccated as above on SS coupons. Following desiccation for 20 days, survivors decreased by 1.4-3.7 log CFU/cm² for non-biofilm L. monocytogenes cells. The contamination level had no significant (p > 0.05) effect on survival kinetics. SEM micrographs showed mature biofilms on coupons initially contaminated with 5.5 and 7.5 log CFU/cm². Mature biofilm cells were significantly (p < 0.05) more desiccation resistant than cells in immature biofilms formed by the lowest contamination level. Besides biofilm maturity/formation, previous osmoadaptation, exposure to lard (20-60%) or salt (5%) during desiccation significantly (p < 0.05) increased the bacterium's survival. In conclusion, L. monocytogenes desiccation survival can be greatly reduced by preventing presence of mature biofilms and salty or fatty soils on food contact surfaces.