• 한국환경농학회:학술대회논문집
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• 2009.07a
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• pp.157-168
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• 2009

Frequent outbreaks of foodborne illness have been increasing the awareness of agro-food safety. Conventional methods for pathogen detection and identification are labor.intensive and take days to complete. The increasing use of rapid food safety testing is receiving more and more attention. The major reason for this trend is that the food industry requires quick and accurate results. The rapid detection of contaminants in food is critical for ensuring the safety of consumers. Recent advances in technology make detection and identification faster, more sensitive and more specific than traditional method. In this paper, technology trends and recent developments in rapid methods for agro-food safety are discussed.

• Genomics & Informatics
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• v.20 no.4
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• pp.41.1-41.9
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• 2022

The pathogen Gallibacterium anatis has caused heavy economic losses for commercial poultry farms around the world. However, despite its importance, the functions of its hypothetical proteins (HPs) have been poorly characterized. The present study analyzed the functions and structures of HPs obtained from Gallibacterium anatis (NCTC11413) using various bioinformatics tools. Initially, all the functions of HPs were predicted using the VICMpred tool, and the physicochemical properties of the identified virulence proteins were then analyzed using Expasy's ProtParam server. A virulence protein (WP_013745346.1) that can act as a potential drug target was further analyzed for its secondary structure, followed by homology modeling and three-dimensional (3D) structure determination using the Swiss-Model and Phyre2 servers. The quality assessment and validation of the 3D model were conducted using ERRAT, Verify3D, and PROCHECK programs. The functional and phylogenetic analysis was conducted using ProFunc, STRING, KEGG servers, and MEGA software. The bioinformatics analysis revealed 201 HPs related to cellular processes (n = 119), metabolism (n = 61), virulence (n = 11), and information/storage molecules (n = 10). Among the virulence proteins, three were detected as drug targets and six as vaccine targets. The characterized virulence protein WP_013745346.1 is proven to be stable, a drug target, and an enzyme related to the citrate cycle in the present pathogen. This enzyme was also found to facilitate other metabolic pathways, the biosynthesis of secondary metabolites, and the biosynthesis of amino acids.

• Journal of Microbiology and Biotechnology
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• v.24 no.6
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• pp.732-739
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• 2014

We describe the development of a polymerase chain reaction method for the rapid, precise, and specific detection of the Xanthomonas oryzae pv. oryzae (Xoo) K3a race, the bacterial blight pathogen of rice. The specific primer set was designed to amplify a genomic locus derived from an amplified fragment length polymorphism specific for the K3a race. The 1,024 bp amplicon was generated from the DNA of 13 isolates of Xoo K3a races out of 119 isolates of other races, pathovars, and Xanthomonas species. The assay does not require isolated bacterial cells or DNA extraction. Moreover, the pathogen was quickly detected in rice leaf 2 days after inoculation with bacteria and at a distance of 8 cm from the rice leaf 5 days later. The results suggest that this PCR-based assay will be a useful and powerful tool for the detection and identification of the Xoo K3a race in rice plants as well as for early diagnosis of infection in paddy fields.

• Journal of Microbiology
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• v.41 no.2
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• pp.144-147
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• 2003

Listeria monocytogenes is a pathogen of major concern to the food industry and the potential cause of severe infections such as listeriosis. Early detection of this foodborne pathogen is important in order to eliminate its potential hazards. So, immunomagnetic separation (IMS) has been suggested as a means of reducing the total analysis time and for improving the sensitivity of detection. Atomic force microscopy (AFM) has been used for measuring the topographic properties of sample surfaces at nanometer scale. In this study, we used AFM to confirm both the sensitivity and the specificity of IMS. Regarding AFM analysis, the length and the width of the bacteria, which were in agreement with literature values, were found to be 2.993 $\mu\textrm{m}$ and 0.837 $\mu\textrm{m}$, respectively. As a result, AFM helped us both characterize and measure the bacterial and bead structures.

• Journal of Forest and Environmental Science
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• v.33 no.4
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• pp.342-347
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• 2017

Since the first report of the oak wilt disease at 2004 in Korea, the disease distributed over Korean peninsula and are still giving severe damages. The management of oak wilt disease in Korea has mainly focused on the control of insect vector, Platypus koryoensis. Neverthless the effective method for evaluating the pathogenicity of the pathogen, Raffaelea quercus-mongolicae (Rqm), and for screening chemical or biological agents with strong inhibitory activity against the pathogen, is absolutely necessary, an reliable method is not available so far. This study was conducted to develop the effective method for evaluating the pathogenicity of Rqm in oak trees. The culture suspensions of Rqm were artificially injected to the saplings of Quercus acutissima by using ChemJet tree injector. Three months after treatments, the treated saplings were cut and dipped into 1% fuchsin acid solution. There were significant differences in non-conductive area (%), discoloration area (%) and vertical discoloration length between the pathogen-injected and distilled water-injected control treatments. These results indicated that the pathogen is the causal agent for the dysfunction of water conductive tissue, which will finally result in wilt symptom. Re-isolation of the pathogen and PCR detection using specific primers for the pathogen also confirmed the presence of Rqm in the sapwood chips of the pathogen-injected saplings. These observations would be greatly applied to other related researches for evaluating the pathogenicity of tree wilt pathogens and biocontrol efficacy of the selected antagonistic microorganisms, in case that the wilt symptom is not easily shown by artificial inoculation of the causal agent.

• Journal of Ginseng Research
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• v.43 no.1
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• pp.1-9
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• 2019

Background: Korean ginseng is an important cash crop in Asian countries. However, plant yield is reduced by pathogens. Among the Ilyonectria radicicola-species complex, I. mors-panacis is responsible for root-rot and replant failure of ginseng in Asia. The development of new methods to reveal the existence of the pathogen before cultivation is started is essential. Therefore, a quantitative real-time polymerase chain reaction method was developed to detect and quantify the pathogen in ginseng soils. Methods: In this study, a species-specific histone H3 primer set was developed for the quantification of I. mors-panacis. The primer set was used on DNA from other microbes to evaluate its sensitivity and selectivity for I. mors-panacis DNA. Sterilized soil samples artificially infected with the pathogen at different concentrations were used to evaluate the ability of the primer set to detect the pathogen population in the soil DNA. Finally, the pathogen was quantified in many natural soil samples. Results: The designed primer set was found to be sensitive and selective for I. mors-panacis DNA. In artificially infected sterilized soil samples, using quantitative real-time polymerase chain reaction the estimated amount of template was positively correlated with the pathogen concentration in soil samples ($R^2=0.95$), disease severity index ($R^2=0.99$), and colony-forming units ($R^2=0.87$). In natural soils, the pathogen was recorded in most fields producing bad yields at a range of $5.82{\pm}2.35pg/g$ to $892.34{\pm}103.70pg/g$ of soil. Conclusion: According to these results, the proposed primer set is applicable for estimating soil quality before ginseng cultivation. This will contribute to disease management and crop protection in the future.

• Parasites, Hosts and Diseases
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• v.56 no.3
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• pp.305-308
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• 2018

This study was aimed to disclose the prevalence rate of tick-borne pathogens from ticks collected from cattle and wild animals in Tanzania in 2012. Ticks were collected from slaughtered cattle and dead wild animals from November 5 to December 23, 2012 and identified. PCR for detecting Anaplasmataceae, Piroplamidae, Rickettsiaceae, Borrelia spp., and Coxiella spp. were done. Among those tested, Rickettsiaceae, Piroplasmidae, and Anaplasmataceae, were detected in ticks from the 2 regions. Rickettsiaceae represented the major tick-borne pathogens of the 2 regions. Ticks from animals in Maswa were associated with a higher pathogen detection rate compared to that in ticks from Iringa. In addition, a higher pathogen detection rate was observed in ticks infesting cattle than in ticks infesting wild animals. All examined ticks of the genus Amblyomma were infected with diverse pathogens. Ticks of the genera Rhipicephalus and Hyalomma were infected with 1 or 2 pathogens. Collectively, this study provides important information regarding differences in pathogen status among various regions, hosts, and tick species in Tanzania. Results in this study will affect the programs to prevent tick-borne diseases (TBD) of humans and livestock in Tanzania.

• Korean Journal of Veterinary Service
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• v.45 no.3
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• pp.145-153
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• 2022

Pathogens such as feline herpesvirus, feline calicivirus, Bordetella bronchiseptica, Chlamydia felis, Mycoplasma felis and Pasteurella multocida usually cause feline upper respiratory tract disease (URTD). Real-time PCR was used to analyze the detection and prevalence of the most common respiratory pathogens in cats with (n=69) and without respiratory signs (n=31). Pathogens were detected in 53 cats, divided into 37 (69.8%) with a single pathogen, 15 (28.3%) with two pathogens, and 1 (1.9%) with three pathogens. M. felis had the highest detection rate in 29 (42.0%) cats, P. multocida was detected in 18 (26.1%), FHV in 10 (14.5%), FCV in 7 (10.1%), B. bronchiseptica in 3 (4.3%), and C. felis in 2 (2.9%). M. felis was the most frequently detected pathogen in cats living outdoors without vaccination. Of the 37 cats infected with single pathogen, nasal discharge was observed in 13 (35.1%), ocular signs in 6 (16.2%), drooling in 5 (13.5%), dyspnea in 3 (8.1%), and asymptomatic in 10 (27.0%). In 51 outdoor and 49 indoor cats, pathogens were detected in 35 (68.6%) and 18 (36.7%) cats, respectively. Of the 29 cats infected with M. felis, 22 (75.9%) showed respiratory signs, and 7 (24.1%) were healthy. In the age of the 53 positive cats, 10 (18.9%) were under the age of 1 year, 26 (49.1%) were aged 1~3 years, and 17 (32.1%) were aged 3 years or older. Although the number of cats in the study was small, the results can provide valuable data on the prevalence of URTD in Korean cats.

• International Journal of Oral Biology
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• v.44 no.1
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• pp.8-13
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• 2019

Recently, the importance of on-site detection of pathogens has drawn attention in the field of molecular diagnostics. Unlike in a laboratory environment, on-site detection of pathogens is performed under limited resources. In this study, we tried to optimize the experimental conditions for on-site detection of pathogens using a combination of ultra-fast convection polymerase chain reaction (cPCR), which does not require regular electricity, and nucleic acid lateral flow (NALF) immunoassay. Salmonella species was used as the model pathogen. DNA was amplified within 21 minutes (equivalent to 30 cycles of polymerase chain reaction) using ultra-fast cPCR, and the amplified DNA was detected within approximately 5 minutes using NALF immunoassay with nucleic acid detection (NAD) cassettes. In order to avoid false-positive results with NAD cassettes, we reduced the primer concentration or ultra-fast cPCR run time. For singleplex ultra-fast cPCR, the primer concentration needed to be lowered to $3{\mu}M$ or the run time needed to be reduced to 14 minutes. For duplex ultra-fast cPCR, $2{\mu}M$ of each primer set needed to be used or the run time needed to be reduced to 14 minutes. Under the conditions optimized in this study, the combination of ultra-fast cPCR and NALF immunoassay can be applied to on-site detection of pathogens. The combination can be easily applied to the detection of oral pathogens.

• The Plant Pathology Journal
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• v.19 no.3
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• pp.133-137
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• 2003

Seedborne fungus Bipolaris spicifera, which has not been previously reported in Korea, was detected from import-ed grass seeds in the country. The most frequently detected fungi from the seeds were Fusarium species, Ulocladium atrum, B. spicifera, Alternaria, and Cuvularia lunata among 17 different seed samples of the family Gramineae. Detection frequencies of B. spicifera were 11,8,5% in Bermuda grass, tall fescue, and mixed lawn grass imported from USA, respectively, and 9％ in mixed lawn grass imported from Italy. This suggests that important seedborne pathogen could be spread between countries through seed sources. The pathogen was seed-transmitted causing damping-off of Bermuda grass seedlings and showed strong pathogenicity to vice, corn, Bermuda grass, sorghum, and tall fescue. However, it did not infect wheat and blue grass.