Jong-Chul Yun

Permanent Genetic Resources added to Molecular Ecology Resources Database 1 December 2009-31 January 2010

This article documents the addition of 220 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Allanblackia floribunda, Amblyraja radiata, Bactrocera cucurbitae, Brachycaudus helichrysi, Calopogonium mucunoides, Dissodactylus primitivus, Elodea canadensis, Ephydatia fluviatilis, Galapaganus howdenae howdenae, Hoplostethus atlanticus, Ischnura elegans, Larimichthys polyactis, Opheodrys vernalis, Pelteobagrus fulvidraco, Phragmidium violaceum, Pistacia vera, and Thunnus thynnus. These loci were cross‐tested on the following species: Allanblackia gabonensis, Allanblackia stanerana, Neoceratitis cyanescens, Dacus ciliatus, Dacus demmerezi, Bactrocera zonata, Ceratitis capitata, Ceratitis rosa, Ceratits catoirii, Dacus punctatifrons, Ephydatia mülleri, Spongilla lacustris, Geodia cydonium, Axinella sp., Ischnura graellsii, Ischnura ramburii, Ischnura pumilio, Pistacia integerrima and Pistacia terebinthus.

Characterization of genes required for the pathogenicity of Pectobacterium carotovorum subsp. carotovorum Pcc21 in Chinese cabbage

Pectobacterium carotovorum subsp. carotovorum is a well-known plant pathogen that causes severe soft rot disease in various crops, resulting in considerable economic loss. To identify pathogenicity-related factors, Chinese cabbage was inoculated with 5314 transposon mutants of P. carotovorum subsp. carotovorum Pcc21 derived using Tn5 transposon mutagenesis. A total of 35 reduced-virulence or avirulent mutants were isolated, and 14 loci were identified. The 14 loci could be functionally grouped into nutrient utilization (pyrD, purH, purD, leuA and serB), production of plant cell-wall-degrading enzymes (PCWDEs) (expI, expR and PCC21_023220), motility (flgA, fliA and flhB), biofilm formation (expI, expR and qseC), susceptibility to antibacterial plant chemicals (tolC) and unknown function (ECA2640). Among the 14 genes identified, qseC, tolC and PCC21_023220 are novel pathogenicity factors of P. carotovorum subsp. carotovorum involved in biofilm formation, phytochemical resistance and PCWDE production, respectively.

Microbiota on Spoiled Vegetables and Their Characterization

Spoilage causes vegetables to deteriorate and develop unpleasant characteristics. Approximately 30 % of fresh vegetables are lost to spoilage, mainly due to colonization by bacteria. In the present study, a total of 44 bacterial isolates were obtained from a number of spoiled vegetables. The isolates were identified and classified into 20 different species of 14 genera based on fatty acid composition, biochemical tests, and 16S rDNA sequence analyses. Pseudomonas spp. were the species most frequently isolated from the spoiled vegetables. To evaluate the spoilage ability of each species, a variety of fresh vegetables were treated with each isolate and their degree of maceration was observed. In addition, the production of plant cell wall-degrading enzymes (PCWDEs), such as cellulase, xylanase, pectate lyase, and polygalacturonase, was compared among isolates to investigate their potential associations with spoilage. Strains that produce more PCWDEs cause spoilage on more diverse plants, and pectinase may be the most important enzyme among PCWDEs for vegetable spoilage. Most gram-negative spoilage bacteria produced acylated homoserine lactone, a quorum-sensing signal molecule, suggesting that it may be possible to use this compound effectively to prevent or slow down the spoilage of vegetables contaminated with diverse bacteria.

Biological Hazard Analysis of Angelica gigas Nakai on Production and Marketing Steps

This study is aimed to investigate microbiological contamination of Angelica gigas Nakai. A total of 111 samples including root, soil, and irrigation water were collected from farms and market to detect aerobic bacteria, Bacillus cereus, coliform, Escherichia coli, Listeria monocytogenes,. Salmonella spp., and Staphylococcus aureus. The contaminations of aerobic bacteria, coliform, and Bacillus cereus in the root during cultivation were found 6.71 log CFU , 4.13 log CFU , and 3.54 log CFU , respectively. The contamination of coliform and B. cereus were detected in all steps from harvesting to processing, with the highest count recorded from the cutting step. In marketing, the contaminations of aerobic bacterial, coliform, and B. cereus were 5.5~6.0 log CFU , 2.4~2.6 log CFU , and 3.5~4.0 log CFU , respectively. Listeria monocytogenes, Salmonella spp, and Staphylococcus aureus were not detected in any of samples. This result indicated that hygienic soil management and post harvest management should be performed to reduce the contamination of hazard microorganisms and to produce safe agro-products.

Investigation of Microbial Contamination Levels of Leafy Greens and Its Distributing Conditions at Different Time - Focused on Perilla leaf and Lettuce -

The objective of this study was to investigate and evaluate microbial contamination levels of leafy greens (perilla leaf and lettuce) and its distributing conditions at different seasons (Feb, May, Aug, and Nov of the year 2011) in order to provide insight into any potential health hazards associated with consumption of these commodities. Leafy greens were collected from a farm located in Geumsan, Chungnam and wholesale markets (WM) and traditional markets (TM) located in Suwon. At the same time, temperature and relative humidity fluctuations experienced by the leafy greens during distribution from the farm to the distribution center were measured by a data logger. The contam- ination levels of perilla leaf and lettuce were determined by analyzing total plate count. Coliform groups, Bacillus cereus, Escherichia coli, Escherichia coli O157:H7, Salmonella spp., Listeria monocytogenes and Staphylococcus aureus were determined. The contamination levels of total aerobic bacteria, coliform groups and B. cereus in both vegetables sampled during May and August found to be higher than those sampled during February and November. E. coli O157:H7, Salmonella spp., L. monocytogenes were not detected in the vegetables analyzed in this study. There were no significant trends between samples at WM and TM in the contamination levels. Relative humidity of vegeta- bles distributed from the farm to the distribution center showed over 90% during distribution regardless of measured seasons. In the case of background microflora on leafy greens, the density was significantly increased at 20, 30 and 37 o C during storage of 24h. E. coli O157:H7 and B. cereus inoculated on the leaves also showed similar increases in the storage tests. The microbial contamination levels determined in this study may be used as the fundamental data for microbial risk assessment.

Prevalence of Pathogenic Bacteria in Livestock Manure Compost and Organic Fertilizer

In recent years, there has been an increasing public concern about fecal contamination of water, air and agricultural produce by pathogens residing in organic fertilizers such as manure, compost and agricultural by-products. Efforts are now being made to control or eliminate the pathogen populations at on-farm level. Development of efficient on-farm strategies to mitigate the potential risk posed by the pathogens requires data about how the pathogens prevail in livestock manure composts and organic fertilizers. Microbiological analysis of livestock manure composts and organic fertilizers obtained from 32 and 28 companies, respectively, were conducted to determine the total aerobic bacteria count, coliforms, Escherichia coli count and the prevalence of Staphylococcus aureus, Bacillus cereus, Salmonella spp., Escherichia coli O157:H7, Listeria monocytogenes, and Cronobacter sakazakii. The total aerobic bacteria counts in the livestock manure composts and organic fertilizers were in the range of 7 to and 4 to , respectively. In the livestock manure composts, coliforms and E. coli were detected in samples obtained from 4 and 2 companies, respectively, in the range of 2 to and . In the organic fertilizers, coliforms and E. coli were detected in samples obtained from 4 and 1 companies, respectively, in the range of 1 to and . In 3 out 32 compost samples, B. cereus was detected, while other pathogens were not detected. In 28 organic fertilizers, no pathogens were detected. The complete composting process can result in the elimination of pathogens in livestock manure compost and organic fertilizer. The results of this study could help to formulate microbiological guidelines for the use of compost in environmental-friendly agriculture. This research provides information regarding microbiological quality of livestock manure compost and organic fertilizer.

Analyses of Microbiological Contamination in Cultivation and Distrubution Stage of Tomato and Evaluation of Microbial Growth in Tomato Extract

This study investigated the microbiological contamination of tomato in cultivation and distribution stage. Growth of Escherichia coli O157:H7 and Listeria monocytogens examined in tomato extracts (0.1, 1.0, and 10.0%) and incubation temperatures (5, 15, 25, and 35 o C). In cultivation stage of tomato, total aerobic bacteria were 7.77 log CFU/g in gloves of APC (Agricultural Products Processing Center) worker and Bacillus cereus were 0.33 log CFU/g at nutrient tank, respectively. And Staphylococcus aureus, Salmonella spp., were not detected. After APC stage, total aerobic bacteria were significantly higher compared with before-APC stage. Among of general, pes- ticide-free and organic produce in tomato were no significant difference in microbial contamination. Coliforms of tomato in small vinyl package were significantly higher when compared to tomato in whole boxes package. There was no significant difference in bacteria count between unwashed tomato and washed tomato using tap water for one minute. The growth of E. coli O157:H7 and L. monocytogens in tomato extracts were decreased significantly as the concentration increased, and the microbial population was reached the lowest point during storage in 10% tomato extracts concentration for 72h at 5 o C. However, the population of E. coli O157:H7 and L. monocytogens were gradu- ally increased at 7.33~8.51 and 7.73~8.60 log CFU/ml during storage at 15~35 o C for 72h, respectively.

Monitoring of Pathogenic Bacteria in Organic Vegetables from Korean Market

This study was undertaken to assess the microbiological quality and prevalence of pathogens in organic vegetables produced in Korea. A total of 189 organically grown vegetable samples (perilla leaf 50, lettuce 50, tomato 39, cucumber 50) were analyzed for the presence of aerobic plate count, Escherichia coli O157:H7, Salmonella spp., Staphylococcus aureus, Listeria monocytogenes, and Yersinia enterocolitica. The total aerobic plate counts were in the range of 4.2 to for perilla leaf, 5.0 to for lettuce, 4.0 to for tomato, and 6.6 to for cucumber. The highest counts were found in cucumber. E. coli O157:H7, Salmonella spp., S. aureus, L. monocytogenes, and Y. enterocolitica were not detected from any organically grown vegetable samples. This research suggests that continuous monitoring in organic vegetables is required to improve fresh produce safety.

Effect of Electrolyzed Water Combined with Ultrasound and Organic Acids to Reduce Salmonella Typhimurium, Staphylococcus aureus, Bacillus cereus on Perilla Leaves

This study was performed to compared the effectiveness of individual treatments (electrolyzed water: EW, organic acid, and ultrasound) and their combination on reducing foodborne pathogens from perilla leaves. Perilla leaves were innoculated with a cocktail of Salmonella Typhimurium, Staphylococcus aureus, Bacillus cereus. Inoculated perilla leaves were treated with EW combined with different concentration of acetic acid (0.5%, 1.0%, 1.5%, 2.0%) for 1 min at room temperature. Treatment of 3 pathogens on perilla leaves with electrolyzed water com- bined with ultrasound (25 kHz) and 0.5% acetic acid was also performed for 1 min. While the numbers of S. Typhimu- rium and B. cereus showed reduced with increasing acetic acid concentration, there is no difference in the number of S. aureus treated with EW containing 0.5% to 1.5% acetic acid. Discoloration was observed the perilla leaves treated with EW combined with more than 1.0% acetic acid. For all three pathogens, the combined treatment of EW and ultra- sound resulted in additional 0.42 to 0.72 log 10 CFU/g. The maxium reductions of S. Typhimurium and B. cereus were 0.95, 1.23 log 10 CFU/g after treatment with EW combined with 0.5% acetic acid and ultrasound simultaneously. The results suggest that the treatment of EW combined with 0.5% acetic acid and ultrasound increased pathogens reduc- tion compared to individual treatment.

Survival and Cross-contamination of Escherichia coli O157:H7 on Various Agricultural Product-Contact Surfaces

To evaluate the effect of surface contaminated with Escherichia coli O157:H7 (E. coli O157:H7) on the microbiological safety of lettuce, this study was conducted to investigate the attachment, biofilm producing, survival, and cross-contamination of E. coli O157:H7 on stainless steel and polyvinyl chloride (PVC). The attachment rate of E. coli O157:H7 on PVC was 10 times higher than that on stainless steel after exposure 1 h in cell suspension. However, there was not a difference between two types of surface after exposure for 6 h and 24h. The biofilm produc- ing of E. coli O157:H7 was TSB > 10% lettuce extracts > 1% lettuce extracts > phosphate buffer. When two kinds of materials were stored at various conditions (20 o C and 30 o C, relative humidity (RH) 43%, 69%, and 100%), the num- bers of E. coli O157:H7 at 30 o C, RH 43% or RH 69% were reduced by 5.0 log CFU/coupon within 12 h regardless of material type. Conversely, the survival of E. coli O157:H7 at RH 100% was lasted more than 5 days. In addition, the reduction rate of E. coli O157:H7 was decreased in the presence of organic matter. The transfer efficiency of E. coli O157:H7 from the contaminated surface to lettuce was dependent upon the water amount of the surface of lettuce. Especially, the transfer rate of E. coli O157:H7 was increased by 10 times in the presence of water on the lettuce sur- face. From this study, the retention of E. coli O157:H7 on produce contact surfaces increase the risk cross-contamina- tion of this pathogen to produce. Thus, it is important that the surface in post harvest facility is properly washed and sanitized after working for prevention of cross-contamination from surface.