• Title/Summary/Keyword: quarantine pathogen

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Development of a Multiplex PCR Method to Detect Fungal Pathogens for Quarantine on Exported Cacti

  • Cho, Hyun ji;Hong, Seong Won;Kim, Hyun-ju;Kwak, Youn-Sig
    • 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.

The pests survey of paprika export complexes and packing house in Korea (우리나라 파프리카 수출단지 및 선과장의 병해충 조사)

  • Kim, Gi-Don;Lee, Siwon;Kang, Eun-Ha;Shin, Yong-Gil;Jeon, Jae-Yong;Heo, Noh-Yeol;Lee, Heung-Sik
    • Korean Journal of Agricultural Science
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    • v.40 no.2
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    • pp.93-99
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    • 2013
  • The disease and insect were surveyed locally in greenhouse, fruit packing house and store house of 51 farms in 13 towns having purpose of paprika exportation. By analysis, various disease and insect were not only founded locally but more ones detected in farms having old facilities and no natural enemy. We found 15 pathogens such as Fusarium spp., Alternaria solani, Leveilluila taurica, PepMV (Pepino mosaic virus) and TMV (Tobacco mosaic virus) in greenhouse, Fusarium spp. in fruit packing house and Penicillium spp. in store house. We found 15 insects in greenhouse such as Bemisia tabaci, rialeurodes vaporariorum and Myzus persicae in greenhouse, Hylobitelus haroldi in fruit packing house. However, the problem quarantine disease and insect for importation and exportation were not detected in inspection time.

Development of Diagnostic Technology of Xylella fastidiosa Using Loop-Mediated Isothermal Amplification and PCR Methods

  • Kim, Suyoung;Park, Yujin;Kim, Gidon
    • Research in Plant Disease
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    • v.27 no.1
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    • pp.38-44
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    • 2021
  • Xylella fastidiosa is the most damaging pathogen in many parts of the world. To increase diagnostic capability of X. fastidiosa in the field, the loop-mediated isothermal amplification (LAMP) and polymerase chain reaction (PCR) assay were developed to mqsA gene of citrate-synthase (XF 1535) X. fastidiosa and evaluated for specificity and sensitivity. Both assays were more robust than current published tests for detection of X. fastidiosa when screened against 16 isolates representing the four major subgroups of the bacterium from a range of host species. No cross reaction with DNA from healthy hosts or other species of bacteria has been observed. The LAMP and PCR assays could detect 10-4 pmol and 100 copies of the gene, respectively. Hydroxynaphthol blue was evaluated as an endpoint detection method for LAMP. There was a significant color shift that signaled the existence of the bacterium when at least 100 copies of the target template were present.

Characterization of Akabane virus (KV0505) from cattle in Korea

  • Yang, Dong-Kun;Kim, Yeon-Hee;Kim, Byoung-han;Kweon, Chang-Hee;Yoon, Soon-Seek;Song, Jae-Young;Lee, Sang-Hun
    • Korean Journal of Veterinary Research
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    • v.48 no.1
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    • pp.61-66
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    • 2008
  • Akabane disease is caused by an arthropod-borne viral pathogen and leads to congenital abnormalities of the central nervous system in infected ruminants. One isolate, KV0505, showed cytopathic effect in Vero cells. The KV0505 isolate was obtained from plasma, which was collected from a cattle raised on Jeju Island in May 2005. Jeju Island is located near the southern part of the Korean peninsula. The isolate was confirmed as Akabane virus (AKAV) by immunofluorescence assay using AKAV specific monoclonal antibodies and reverse transcription polymerase chain reaction (RTPCR). Suckling mice inoculated with the isolate showed signs of paralysis and died within 10 days postinoculation. Comparisons of the KV0505 N gene sequence with 39 other known AKAV strains revealed nucleotide homologies ranging from 83.6% (MP496 strain) to 99.7% (M171 strain). When compared with the K-9 strain, which was isolated from a cow in Korea in 1994, the nucleotide sequence homology with the N gene was 99.7%. Thus, genes of the KV0505 isolate were closely related to those of the M171 strain, which were clustered into the Ic group of AKAV.

Identification and Molecular Analysis of Ixodid Ticks (Acari: Ixodidae) Infesting Domestic Animals and Tick-Borne Pathogens at the Tarim Basin of Southern Xinjiang, China

  • Zhao, Li;Lv, Jizhou;Li, Fei;Li, Kairui;He, Bo;Zhang, Luyao;Han, Xueqing;Wang, Huiyu;Johnson, Nicholas;Lin, Xiangmei;Wu, Shaoqiang;Liu, Yonghong
    • Parasites, Hosts and Diseases
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    • v.58 no.1
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    • pp.37-46
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    • 2020
  • Livestock husbandry is vital to economy of the Tarim Basin, Xinjiang Autonomous Region, China. However, there have been few surveys of the distribution of ixodid ticks (Acari: Ixodidae) and tick-borne pathogens affecting domestic animals at these locations. In this study, 3,916 adult ixodid ticks infesting domestic animals were collected from 23 sampling sites during 2012-2016. Ticks were identified to species based on morphology, and the identification was confirmed based on mitochondrial 16S and 12S rRNA sequences. Ten tick species belonging to 4 genera were identified, including Rhipicephalus turanicus, Hyalomma anatolicum, Rh. bursa, H. asiaticum asiaticum, and Rh. sanguineus. DNA sequences of Rickettsia spp. (spotted fever group) and Anaplasma spp. were detected in these ticks. Phylogenetic analyses revealed possible existence of undescribed Babesia spp. and Borrelia spp. This study illustrates potential threat to domestic animals and humans from tick-borne pathogens.

Potential Pathogen Monitoring of Powdered Infant Formula Milk and Related Products in Korea (국내산 조제유류에서의 위해 미생물 모니터링)

  • Kim, Young-Jo;Moon, Jin-San;Park, Hyun-Jung;Heo, Eun-Jeong;Kim, Ji-Ho;Lee, Hee-Soo;Wee, Sung-Hwan
    • Journal of Food Hygiene and Safety
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    • v.25 no.4
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    • pp.341-345
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    • 2010
  • Three-hundred samples of powdered infant formula milk and related products from four different manufacturers in 2010 were collected and surveyed their contaminations for aerobic bacteria, coliform, Enterobacter(Cronobacter) sakazakii, and food-borne pathogens. Fifteen samples of sterilized infant formula milk were all negative on these microorganisms. In all collected products of un sterilized infant formulas and follow-on infant formulas, aerobic bacteria were detected at 239 (83.9%) among 285 samples, and they all were found below $10^3$ cfu/g. Coliform bacteria were also detected at four among 285 samples. Salmonella spp. and Ent. sakazakii, weren't detected at the all samples. Bacillus cereus was detected at 24 (8.4%) among 285 samples. The level of B. cereus was below 100 cfu/g but it was suitable for the range of specification of B. cereus in infant formulas. Clostridium perjringens, Escherichia coli O157:H7, Staphylococcus aureus and Listeria monocytogenes weren't also detected. In consequence, it was suitable for total viable count, coliform and potential pathogen to the specification of infant formulas and related products.