The finding that C9-deficient sera (C9D) can kill serum sensitive strains of Gram-negative bacteria by us and other investigators, questions the role of C9 in the membrane attack complex as necessary for cell death. In these studies we have demonstrated that C5b-8 complexes generated on E. coli J5 during incubation in C9-depleted and C9-neutralized sera are effective in killing Gram-negative bacteria. In the same study, we extended our investigations to show that the deposition of C5b-7 complexes (from C8-deficient [C8D], C8 depleted and C8-neutralized sera) is also effective in killing Gram-negative bacteria. In all cases, these studies demonstrated that when E. coli J5 was incubated with C8D, C9D and pooled normal human serum [PNHS], deposited C5b-9 complexes from PNHS produced more killing than C5b-7 or C5b-8 complexes alone. These experiments clearly demonstrated that C5b-7 and C5b-8 complexes are bactericidal and that multimeric C9 within C5b-9 is not an absolute requirement for inner membrane damage and cell death of Gram-negative bacteria.

Efficacy of lactic acid bacteria (LAB) Lactobacillus plantarum and Pediococcus pentosaceus for reducing Listeria on cantaloupes was investigated. Cantaloupe surfaces inoculated with Listeria innocua and Listeria monocytogenes in preharvest and postharvest studies, respectively, were treated with or without LAB. Listeria populations were enumerated on days 0, 1, 5, and 7. At the preharvest level, treatment with unwashed LAB (cultures grown in De Man, Rogosa, and Sharpe) significantly reduced L. innocua by 2.1 log and 1.5 log CFU/cm2 on cantaloupes harvested on days 5 and 7, respectively. At the postharvest level, both unwashed LAB and washed LAB (cultures resuspended in phosphate buffer saline) significantly reduced L. monocytogenes by >2.0 log CFU/cm2 on cantaloupes after 7 days of storage at 25°C; the unwashed LAB treatment exerted higher anti‐Listeria efficacy than the washed LAB. Results suggest that these LAB strains offer potential application for reducing Listeria contamination on cantaloupes at preharvest and postharvest levels. Practical applications Listeriosis associated with the consumption of cantaloupes have resulted in 153 illnesses, 148 hospitalizations, and 33 deaths in the United States between 2009 and 2013, which highlight the need for effective interventions to improve cantaloupe safety. Cantaloupes may be contaminated with Listeria at the farm level or during wash step at the packing facility. Lactic acid bacteria (LAB) including Lactobacillus plantarum and Pediococcus pentosaceus can be potentially utilized as biocontrol agents on the surface of the cantaloupes at the farm level or during postharvest wash to reduce the populations of Listeria on cantaloupes. Further, LAB may minimize cross‐contamination of Listeria at the postharvest level.

Treated wastewater (TW) and roof-collected rain water (RW) that meet the required microbial quality as per Food Safety Modernization Act (FSMA) regulation may serve as alternative irrigation water sources to decrease the pressure on the current water scarcity. Alternative water sources may have different water characteristics that influence the survival and transfer of microorganisms to the irrigated produce. Further, these water sources may contain pathogenic bacteria such as Shiga-toxigenic Escherichia coli. To evaluate the risk associated with TW and RW irrigation on the fresh produce safety, the effect of TW and RW irrigation on the transfer of two non-pathogenic E. coli strains as surrogates for E. coli O157:H7 to different lettuce cultivars grown in the field was investigated. Lettuce cultivars “Annapolis,” “Celinet,” and “Coastline” were grown in the field at the Fulton farm (Chambersburg, PA). Approximately 10 days before harvest, lettuce plants were spray-irrigated with groundwater (GW), TW, or RW containing 6 log CFU ml−1 of a mixture of nalidixic acid-resistant E. coli O157:H12 and chloramphenicol-resistant E. coli K12 in fecal slurry as non-pathogenic surrogates for E. coli O157:H7. On 0, 1, 3, 7, and 10 days post-irrigation, four replicate lettuce leaf samples (30 g per sample) from each group were collected and pummeled in 120 ml of buffered peptone water for 2 min, followed by spiral plating on MacConkey agars with antibiotics. Results showed that the recovery of E. coli O157:H12 was significantly greater than the populations of E. coli K12 recovered from the irrigated lettuce regardless of the water sources and the lettuce cultivars. The TW irrigation resulted in the lowest recovery of the E. coli surrogates on the lettuce compared to the populations of these bacteria recovered from the lettuce with RW and GW irrigation on day 0. The difference in leaf characteristics of lettuce cultivars significantly influenced the recovery of these surrogates on lettuce leaves. Populations of E. coli O157:H12 recovered from the RW-irrigated “Annapolis” lettuce were significantly lower than the recovery of this bacterium from the “Celinet” and “Coastline” lettuce (P < 0.05). Overall, the recovery of specific E. coli surrogates from the RW and TW irrigated lettuce was comparable to the lettuce with the GW irrigation, where GW served as a baseline water source. E. coli O157:H12 could be a more suitable surrogate compared to E. coli K12 because it is an environmental watershed isolate. The findings of this study provide critical information in risk assessment evaluation of RW and TW irrigation on lettuce in Mid-Atlantic area.

Listeria monocytogenes and Shiga toxigenic Escherichia coli (STEC) are important foodborne bacterial pathogens that can form biofilms on equipment surfaces at food processing facilities. Pathogens in biofilms are resistant to conventional antimicrobials and require higher antimicrobial concentrations to be inactivated. In this study, the efficacy of a synthetic innate defense regulator peptide 1018 (peptide 1018) for inactivating L. monocytogenes and STEC (O26, O111, O145, O157) biofilms on stainless steel and polycarbonate surfaces was investigated. Stainless steel and polycarbonate coupons (12 mm in diameter) were used in a Centers for Disease Control and Prevention biofilm reactor containing 400 mL of 10% tryptic soy broth (TSB) that had been inoculated with an individual strain of L. monocytogenes or STEC to obtain 6 log CFU/mL populations. The reactor was set with a constant flow rate at 50 mL/h of 10% TSB for 48 h. After 48 h, coupons were treated with peptide 1018 at 0, 10, 20, or 50 μg/mL in phosphate buffer saline (PBS) for 24 h. Surviving bacterial populations were determined by scraping off the coupons and spiral plating on selective media. Significantly higher levels of pathogens in biofilms formed by certain bacterial strains, including L. monocytogenes F6854, E. coli O157:H7 RM4407 and NADC5713, and non-O157 E. coli NADC3629, were recovered on polycarbonate surfaces than on stainless steel. Antibiofilm efficacy of peptide 1018 against pathogens was concentration-dependent and varied with the type of pathogen and material surfaces. Peptide 1018 at 50 μg/mL significantly inactivated all tested bacterial biofilms on both surfaces compared with the PBS control (P < 0.05). L. monocytogenes was the bacterium most sensitive to peptide 1018; on stainless steel surfaces treated with 50 μg/mL peptide 1018, there was a 3.7- to 4.6-log CFU/cm2 reduction in Listeria populations compared with a 1.0- to 3.5-log CFU/cm2 reduction of STEC. Results suggest that peptide 1018 may be used to inactivate L. monocytogenes and STEC biofilms on equipment surfaces.