• Research in Plant Disease
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• v.22 no.4
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• pp.243-248
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• 2016

As preventive measures for bonsai exports, nematodes were isolated from 55 bonsai samples of five coniferous species (Chamaecyparis pisifera, Juniperus chinensis, Pinus densiflora, Pinus thunbergii, and Taxus cuspidate) from all 15 bonsai gardens in Korea. Nematodes belonging to 21 genera in 7 orders were isolated from the 55 bonsai samples. Among plant-parasitic nematodes, Tylenchus spp. was the most frequently isolated (14.9%), followed by Ditylenchus spp. (10.5%), Aphelenchoides spp. (9.5%), Aphelenchus sp. (5.5%), Criconemoides sp. (4.0%), Helicotylenchus sp. (0.7%), Hemicycliophora sp. (0.7%), Mesocriconema sp. (0.7%), Tylenchorhynchus sp. (0.7%), and Paratylenchus sp. (0.4%). Among nonparasitic nematodes, Cephalobina was the most frequently isolated nematodes (26.5%), followed by Rhabditida (19.3%), Dorylaimida (17.8%), Pangrolaimida (14.5%), Plectida (6.5%), Tryphylida (6.2%), Mononchida (3.3%), Alaimida (2.9%), Monhysterida (2.5%), and Triplonchida (0.4%). Based on these results, we conclude that there is no problematic plant-parasitic nematode in bonsai gardens of Korea.

• The Plant Pathology Journal
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• v.27 no.4
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• pp.367-371
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• 2011

Multiplex PCR assays were developed for the simultaneous detection of ten important Korean quarantine phytoplasmas. The species-specific primers were designed based on ribosomal protein, putative preprotein translocase Y, immunodominant protein, elongation factor TU, chaperonin protein and the 16S rRNA genes of 'Candidatus (Ca.) Phytoplasma' species. Three main primer sets were prepared from ten designed primer pairs to limit nonspecific amplification as much as possible. The multiplex PCR assay using the three primer sets successfully amplified the correct conserved genes for each 'Ca. Phytoplasma' species. In addition, ten important 'Ca. Phytoplasma' species could be easily determined by recognizing band patterns specific for each phytoplasma species from three primer sets. Moreover, a high sensitivity of multiplex PCR for each primer set was observed for samples containing a low DNA concentration (10 ng/${\mu}l$). This study provides the useful multiplex PCR assay as a convenient method to detect the presence of ten important quarantine phytoplasmas in Korea.

• Food Science of Animal Resources
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• v.33 no.2
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• pp.239-243
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• 2013

Salmonella Enteritidis is responsible for causing foodborne diseases upon consumption of egg products. While cases of S. Enteritidis isolation from eggs have been reported in other countries, no such cases have previously been reported in Korea. In this study, we report the first isolation and identification of S. Enteritidis from domestically distributed eggs in Korea. Eggs were collected from eight countrywide grocery stores during different seasons between 2011 and 2012. Egg contents and washing solution of egg shells were incubated in buffered peptone water, and the enriched broth was further enriched in tetrathionate broth and Rappaport-Vassiliadis. The secondary enriched broth was streaked on xylose lysine desoxycholate agar. The suspected colonies were confirmed to S. Enteritidis by a biochemical test, serotyping, and PCR test. Genetic relatedness among the isolates was analyzed using Diversilab Salmonella kit. Three strains of S. Enteritidis were isolated from egg contents and egg shells collected from grocery stores of the Eumseong-city in the fall of 2011. All three stains showed resistance to chloramphenicol, streptomycin, nalidixic acid, and ampicillin by the disk diffusion method. In addition, the isolates showed more than 99% DNA homology, indicating that they were presumably identical strains. Therefore, there is a requirement to monitor and control against S. Enteritidis from eggs in Korea.

• Microbiology and Biotechnology Letters
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• v.43 no.3
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• pp.296-299
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• 2015

Cowpea mild mottle virus (CPMMV) has a wide range of hosts, such as the pea family and tomato. CPMMV is a non-reported virus in Korea, and is domestically designated as a controlled virus associated with plant quarantine. In this study, a rapid diagnostic method for the detection of CPMMV at quarantine sites was developed. For the development of a user-based system, the PCR compositions and conditions use existing methods of quarantine for the viruses. Two sets of RT-PCR and nested PCR were developed in this study that could be amplified from 579 → 298 dp and 638 → 252 bp, respectively. Furthermore, a sequence inserted positive control plasmid was developed, which is able to identify false-positives resulting from laboratory contamination. The findings of this study are important for the diagnosis of CPMMV in imported crops held in plant quarantine.

• Parasites, Hosts and Diseases
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• v.52 no.1
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• pp.57-62
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• 2014

Biting midges (Culicoides: Ceratopogonidae) were collected by Mosquito Magnet$^{(R)}$ traps at the Neutral Nations Supervisory Commission (NNSC) camp and Daeseongdong village inside the demilitarized zone (DMZ) and near the military demarcation line (MDL) separating North and South Korea and at Warrior Base (US Army training site) and Tongilchon 3 km south of the DMZ in northern Gyeonggi Province, Republic of Korea (ROK), from May-October 2010-2012, to determine their seasonal distributions. A total of 18,647 Culicoides females (18,399; 98.7%) and males (248; 1.3%) comprising 16 species were collected. Overall, the most commonly collected species was Culicoides nipponensis (42.9%), followed by C. erairai (29.2%), C. punctatus (20.3%), C. arakawae (3.3%), C. pallidulus (1.8%), and C. circumscriptus (1.4%), while the remaining 10 species accounted for only 1.1% of all Culicoides spp. collected. The seasonal distribution of C. nipponensis was bimodal, with high numbers collected during May-June and again during September. C. erairai was more frequently collected during June-July, followed by sharply decreased populations from August-October. C. punctatus was collected in low numbers from May-September with high numbers collected during October. C. erairai was predominantly collected from the NNSC camp (85.1% of all C. erairai collected) located adjacent to the MDL at Panmunjeom in the northernmost part of Gyeonggi-do (Province), while other sites yielded low numbers of specimens.

• Animal Systematics, Evolution and Diversity
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• v.36 no.1
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• pp.41-45
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• 2020

The genus Philanthus Fabricius, 1790 in Korea are reviewed, and two species, Philanthus triangulum and P. coronata, are treated. The former is new to Korea. This species is easily separated from congeners by dense punctures in propodral enclosure and tri-forked marking in lower face. DNA barcoding test is supportive in our identification as well as conspecitficity of Korean materials showing variation in size and coloration. The observed data on flower associations of this species specified with domestic localites and date are separately provided. The current status of Philanthus coronatus that has been the only representative of the genus but forgotten for a lengthy time in Korea is discussed.

• Journal of Food Hygiene and Safety
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• v.31 no.1
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• pp.28-35
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• 2016

Species identification of animal tissues in meat products is an important issue to protect the consumer from illegal and/or undesirable adulteration; for economic, religious and health reasons. In this reason, accurate analytical methods are needed for the labeling of meat products with requiring simple and fast procedure. Recently, applications of PCR in food analysis have been increased because of their simplicity, specificity and sensitivity. Therefore, in this study, a multiplex PCR assay was developed for the simultaneous identification of eight species of cow, pig, chicken, duck, goat, sheep, horse and turkey from raw meats. The primers were designed in different regions of mitochondrial 16S RNA after alignment of the available sequences in the GenBank database. Two multiplex primer sets were designed as Set 1 (cow, pig, chicken, duck) and Set 2 (goat, sheep, horse, turkey), respectively. Total 274 samples from cow (n = 55), pig (n = 30), chicken (n=30), and duck (n = 30), goat (n = 40), sheep (n = 33), horse (n = 41), and turkey (n = 15) were tested. The primers generated specific fragments of 94, 192, 279, 477 bp (pig, chicken, cow, duck), 670, 271, 152, 469 bp (goat, sheep, horse, turkey) lengths for eight species, respectively. The animal species specificity was 100% in all eight samples in the multiplex PCR assay. The detection limit of the multiplex PCR assay showed from 100 fg to 1 pg of template DNA from extracted from raw meats. When applying multiplex PCR assays to sample from pork/beef and pork/chicken, beef/chicken tested raw mixed meats and heat-treated ($83^{\circ}C$ for 30min, $100^{\circ}C$ for 20min, and $121^{\circ}C$ for 10min) mixtures, detection limit was 0.1% level beef, pork and pork in beef and chicken in pork and 1.0% level pork in chicken. This study suggest that the developed multiplex PCR assay can be used for rapid and simultaneous species identification of cow, pig, chicken, duck, goat, sheep, horse and turkey from meats.

• The Plant Pathology Journal
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• v.32 no.1
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• pp.53-57
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• 2016

Major diseases in grafted cacti have been reported and Fusarium oxysporum, Bipolaris cactivora, Phytophthora spp. and Collectotrichum spp. are known as causal pathogens. These pathogens can lead to plant death after infection. Therefore, some European countries have quarantined imported cacti that are infected with specific fungal pathogens. Consequently, we developed PCR detection methods to identify four quarantined fungal pathogens and reduce export rejection rates of Korean grafted cacti. The pathogen specific primer sets F.oF-F.oR, B.CF-B.CR, P.nF-P.nR, and P.cF-P.CR were tested for F. oxysporum, B.cactivora, P. nicotinae, and P. cactorum, respectively. The F.oF-F.oR primer set was designed from the Fusarium ITS region; the B.CF-B.CR and P.nF-P.nR primers respectively from Bipolaris and Phytophthora ITS1; and the P.cF-P.CR primer set from the Ypt1protein gene region. The quarantine fungal pathogen primer pairs were amplified to the specific number of base pairs in each of the following fungal pathogens: 210-bp (F. oxysporum), 510-bp (B. cactivora), 313-bp (P. nicotinae), and 447-bp (P. cactorum). The detection limit for the mono- and multiplex PCR primer sets was 0.1 ng of template DNA under in vitro conditions. Therefore, each primer set successfully diagnosed contamination of quarantine pathogens in export grafted cacti. Consequently, our methodology is a viable tool to screen contamination of the fungal pathogen in exported grafted cacti.

• International Journal of Industrial Entomology and Biomaterials
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• v.36 no.2
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• pp.42-48
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• 2018

We found the large narcissus fly, Merodon equestris (Fabricius), which has been doubted to occur in Korea. This species is an economically important in management of narcissus and also of quarantine pests. We therefore provide the morphological diagnosis and DNA barcode sequences for rapid species identification of M. equestris based on the five Korean specimens.

• Journal of Veterinary Science
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• v.22 no.3
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• pp.40.1-40.12
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• 2021

Background: The microsporidian parasite Nosema ceranae is a global problem in honeybee populations and is known to cause winter mortality. A sensitive and rapid tool for stable quantitative detection is necessary to establish further research related to the diagnosis, prevention, and treatment of this pathogen. Objectives: The present study aimed to develop a quantitative method that incorporates ultra-rapid real-time quantitative polymerase chain reaction (UR-qPCR) for the rapid enumeration of N. ceranae in infected bees. Methods: A procedure for UR-qPCR detection of N. ceranae was developed, and the advantages of molecular detection were evaluated in comparison with microscopic enumeration. Results: UR-qPCR was more sensitive than microscopic enumeration for detecting two copies of N. ceranae DNA and 24 spores per bee. Meanwhile, the limit of detection by microscopy was 2.40 × 10⁴ spores/bee, and the stable detection level was ≥ 2.40 × 10⁵ spores/bee. The results of N. ceranae calculations from the infected honeybees and purified spores by UR-qPCR showed that the DNA copy number was approximately 8-fold higher than the spore count. Additionally, honeybees infected with N. ceranae with 2.74 × 10⁴ copies of N. ceranae DNA were incapable of detection by microscopy. The results of quantitative analysis using UR-qPCR were accomplished within 20 min. Conclusions: UR-qPCR is expected to be the most rapid molecular method for Nosema detection and has been developed for diagnosing nosemosis at low levels of infection.