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Outbreaks of Salmonellosis have been traced to contaminated tomato. The produce production environment poses a risk for Salmonella contamination; however, little is known about the effects of pest management practices on Salmonella during production. The study objective was to evaluate pesticide application on the inactivation of Salmonella on tomato leaves. Thirty greenhouse‐grown tomato plants were inoculated with S. enterica serovars Newport or Typhimurium. Inoculation was performed by dipping tomato leaves in an 8‐log CFU/mL Salmonella suspension with 0.025% (vol/vol) Silwet L‐77 surfactant for 30 s, for a starting concentration of 6–7 log CFU/mL. Plants were treated with one of four pesticides, each with a different mode of action [acibenzolar‐S‐methyl, copper‐hydroxide, peroxyacetic acid (PAA), and streptomycin]. Pesticides were applied at manufacturers' labeled rate for plant disease management with water as a control treatment. Salmonella was enumerated at 0.125 (3 h), 2, 6, and 9 days post‐inoculation (dpi), and counts log‐transformed. Growth of Salmonella was not observed. At 2 dpi, PAA and streptomycin significantly reduced surface Salmonella concentrations of inoculated tomato leaves (0.7 and 0.6‐log CFU/g, respectively; p ≤ 0.05), while significant Salmonella log reduction occurred in the ground tomato leaves after copper hydroxide treatment (0.8‐log CFU/g; p ≤ 0.05), compared to the control. No significant differences in Salmonella populations on tomato leaf surface and in ground leaves were observed from 2 to 9 dpi, regardless of pesticide application. These findings suggest single in‐field pesticide applications may not be an effective mitigation strategy in limiting potential Salmonella contamination. Future research, including multiple in‐field pesticide applications, or pesticide use in combination with other mitigation strategies, may offer intriguing management practices to limit possible preharvest contamination. [ABSTRACT FROM AUTHOR]

Outbreaks of foodborne illnesses linked to fresh fruits and vegetables have been key drivers behind a wide breadth of research aiming to fill data gaps in our understanding of the total ecology of agricultural water sources such as ponds and wells and the relationship of this ecology to foodborne pathogens such as Salmonella enterica and Listeria monocytogenes. Both S. enterica and L. monocytogenes can persist in irrigation water and have been linked to produce contamination events. Data describing the abundance of these organisms in specific agricultural water sources are valuable to guide water treatment measures. Here, we profiled the culture independent water microbiota of four farm ponds and wells correlated with microbiological recovery of S. enterica (prevalence: pond, 19.4%; well, 3.3%), L. monocytogenes (pond, 27.1%; well, 4.2%) and fecal indicator testing. Correlation between abiotic factors, including water parameters (temperature, pH, conductivity, dissolved oxygen percentage, oxidation reduction potential, and turbidity) and weather (temperature and rainfall), and foodborne pathogens were also evaluated. Although abiotic factors did not correlate with recovery of S. enterica or L. monocytogenes (p > 0.05), fecal indicators were positively correlated with incidence of S. enterica in well water. Bacterial taxa such as Sphingomonadaceae and Hymenobacter were positively correlated with the prevalence and population of S. enterica , and recovery of L. monocytogenes was positively correlated with the abundance of Rhizobacter and Comamonadaceae (p < 0.03). These data will support evolving mitigation strategies to reduce the risk of produce contamination by foodborne pathogens through irrigation. [ABSTRACT FROM AUTHOR]

Vibrio parahaemolyticus (Vp) is a common marine halophilic food‐borne pathogen, mainly found in seafood and food with a high salt content. Gastrointestinal reactions such as diarrhea, headache, vomiting, nausea, and abdominal cramps may occur after eating food infected with Vp. This study aimed to screen for high‐affinity aptamers that specifically recognize Vp. A high‐affinity modified aptamer screening kit was used to rapidly screen aptamers of the food‐borne Vp. The first round of screening involved release of target aptamers from the microspheres. The "false‐positive" aptamers were eliminated after specific binding to and elution of Vp in the second round. The second round of screening of the aptamers involved polymerase chain reaction (PCR), and the abundance of a sequence was determined using next‐generation sequencing. Nine high‐affinity aptamer sequences were obtained, and the first eight modified aptamer sequences were derived using a cloud‐based intelligent software of the American AM Biotech Co. Escherichia coli (E. coli) was used as a control, and aptamer ID 12 with the highest affinity for Vp was selected using real‐time PCR. According to the principle of color change caused by nano‐gold condensing under salt induction, Salmonella, Listeria monocytogenes (L. monocytogenes), and E. coli were used as counter‐screening bacteria, and the aptamer ID12 was combined with nano‐gold. The results showed that aptamer ID12 has strong specificity for Vp. Based on these findings, this study developed a simple, innovative, and rapid method for screening Vp aptamers. [ABSTRACT FROM AUTHOR]

The Eastern Shore of Virginia (ESV) is a major agricultural region in Virginia and in the past has been linked to some tomato-associated outbreaks of salmonellosis. In this study, water samples were collected weekly from irrigation ponds and wells in four representative vegetable farms (Farms A–D, each farm paired with one pond and one well) and a creek as well. In addition, water samples from two sites in the Chesapeake Bay on the ESV were collected monthly. Poultry litter was sampled monthly from three commercial broiler farms. Soil samples were collected monthly after fertilization with poultry litter from 10 farms in 2014 and another 14 farms in 2015. A most probable number method was used to detect Salmonella enterica presence and concentration in collected samples. Presumptive Salmonella colonies were confirmed by the cross-streaking method. Molecular serotyping was carried out to determine the Salmonella serovars. The average prevalence of Salmonella in pond, well, creek, and bay water samples was 19.3, 3.3, 24.2, and 29.2%, respectively. There were significant spatial and temporal differences for Salmonella incidence in various water sources. The prevalence of S. enterica in four tested ponds from farms A, B, C, and D were 16, 12, 22, and 27%, respectively. While the prevalence of S. enterica in irrigation wells was significantly lower, some well water samples tested positive during the study. Salmonella Newport was found to be the predominant serovar isolated from water samples. All poultry houses of the three tested broiler farms were Salmonella -positive at certain sampling points during the study with prevalence ranging from 14.3 to 35.4%. Salmonella was found to be able to survive up to 4 months in poultry litter amended soils from the tested farms in 2014, and up to 6 months in 2015. This research examined the dynamics of S. enterica in relationship to water source, poultry litter, and amended soil in a major agricultural area, and provides useful information for food safety risk assessments. [ABSTRACT FROM AUTHOR]

Between 2000 and 2010 the Eastern Shore of Virginia was implicated in four Salmonella outbreaks associated with tomato. Therefore, a multi-year study (2012–2015) was performed to investigate presumptive factors associated with the contamination of Salmonella within tomato fields at Virginia Tech's Eastern Shore Agricultural Research and Extension Center. Factors including irrigation water sources (pond and well), type of soil amendment: fresh poultry litter (PL), PL ash, and a conventional fertilizer (triple superphosphate – TSP), and production practices: staked with plastic mulch (SP), staked without plastic mulch (SW), and non-staked without plastic mulch (NW), were evaluated by split-plot or complete-block design. All field experiments relied on naturally occurring Salmonella contamination, except one follow up experiment (worst-case scenario) which examined the potential for contamination in tomato fruits when Salmonella was applied through drip irrigation. Samples were collected from pond and well water; PL, PL ash, and TSP; and the rhizosphere, leaves, and fruits of tomato plants. Salmonella was quantified using a most probable number method and contamination ratios were calculated for each treatment. Salmonella serovar was determined by molecular serotyping. Salmonella populations varied significantly by year; however, similar trends were evident each year. Findings showed use of untreated pond water and raw PL amendment increased the likelihood of Salmonella detection in tomato plots. Salmonella Newport and Typhimurium were the most frequently detected serovars in pond water and PL amendment samples, respectively. Interestingly, while these factors increased the likelihood of Salmonella detection in tomato plots (rhizosphere and leaves), all tomato fruits sampled (n = 4800) from these plots were Salmonella negative. Contamination of tomato fruits was extremely low (< 1%) even when tomato plots were artificially inoculated with an attenuated Salmonella Newport strain (104 CFU/mL). Furthermore, Salmonella was not detected in tomato plots irrigated using well water and amended with PL ash or TSP. Production practices also influenced the likelihood of Salmonella detection in tomato plots. Salmonella detection was higher in tomato leaf samples for NW plots, compared to SP and SW plots. This study provides evidence that attention to agricultural inputs and production practices may help reduce the likelihood of Salmonella contamination in tomato fields. [ABSTRACT FROM AUTHOR]

Ammonia-oxidizing archaea (AOA) have more importance in ammonia oxidation than ammonia-oxidizing bacteria (AOB) in acidic red soils. The aim of this study was to investigate if the abundance and composition of AOA could be altered by long-term application of organic manure in an acidic red soil. The abundance and composition of AOA were evaluated by polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE) targeting archaealamoAgenes after long-term (24-year) application of mineral fertilizer and/or organic manure. The treatments were: non-fertilized control, mineral nitrogen (N) fertilizer only, mineral N, phosphorus (P) and potassium (K) fertilizer only, organic manure only, and organic manure plus mineral NPK fertilizer. The abundance of archaealamoAgenes was significantly increased after the long-term application of organic manures, either with or without mineral NPK fertilizer. So were the Shannon and Richness diversity indices of AOA deduced from the DGGE patterns. Phylogenetic analyses showed that most of the AOA sequences from various fertilization treatments were affiliated with group 1.1b thaumarchaea and only one with the group 1.1a-associated thaumarchaea. Nitrification potential was significantly increased after the long-term application of organic manures in comparison with the non-fertilized control. Our results strengthened the importance of organic manure in promoting the growth of AOA and thus nitrification potential in the acidic red soils. [ABSTRACT FROM AUTHOR]

Copyright of Canadian Journal of Microbiology is the property of Canadian Science Publishing and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Canadian Journal of Microbiology is the property of Canadian Science Publishing and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Internal contamination of Salmonella in plants is attracting increasing attention for food safety reasons. In this study, three different tomato cultivars "Florida Lanai", "Crown Jewel", "Ailsa Craig" and the transgenic line Sp5 of "Ailsa Craig" were inoculated with 1 ml GFP-labeled Salmonella Typhimurium through guttation droplets at concentrations of 109 or 107 CFU/ ml. Survival of Salmonella on/in tomato leaves was detected by both direct plating and enrichment methods. Salmonella cells survived best on/in the inoculated leaves of cultivar "Ailsa Craig" and decreased fastest on/in "Florida Lanai" leaves. Increased guttation in the abscisic acid over-expressing Sp5 plants may have facilitated the entrance of Salmonella into leaves and the colonization on the surface of tomato leaves. Internalization of Salmonella Typhimurium in tomato leaves through guttation drop inoculation was confirmed by confocal laser microscopy. For the first time, convincing evidence is presented that S. enterica can enter tomato leaves through hydathodes and move into the vascular system, which may result in the internal translocation of the bacteria inside plants. [ABSTRACT FROM AUTHOR]

Burkholderia contaminans strain MS14 has a broad range of antifungal activities to plant and human pathogens. In previous studies, a 22.7-kb genomic fragment harboring six genes was shown to be involved in the production of an antifungal oligopeptide in B. contaminans strain MS14. In this study, another LuxR-type regulatory gene, named ambR1, was identified downstream of the ambR2 gene, and three new ORFs were found upstream of ORF6 of the 22.7-kb fragment. Site-directed mutagenesis revealed that ambR1 was required for expression of the antifungal activity against the indicator fungus Geotrichum candidum. Transcription of all the putative genes (ORFs 2–9) identified in the region except ORF1 was regulated by both ambR1 and ambR2. The functional ambR1 gene was essential for transcription of ambR2, and constitutive expression of ambR2 did not restore the phenotype of the mutant MS14GG44( ambR1∷ nptII). Two of the three ORFs identified upstream from the ORF6 were predicted to encode two nonribosomal peptide synthetases (ORF7 and ORF9), and an insertion mutation in ORF9 resulted in the loss of antifungal activity against G. candidum. These results suggest that ambR1 is the key regulatory gene controlling the production of the antifungal activity of strain MS14. [ABSTRACT FROM AUTHOR]