A total of 501 serum samples were selected from blood samples that were submitted to Department of Veterinary Pathology, Kangwon National University from all provinces in Korea from September 2001 to August 2002. Their sera were examined for antibodies to swine influenza virus subtype H1N1 (SlV H1N1) and porcine repro-ductive and respiratory syndrome virus (PRRSV) according to the age of pig, season, and herd size using enzyme-linked immunosorbent assay. The seroprevalence of SIV H1N1, PRRSV, and dual infection were 39.12%, 61.48%, and 25.95%, respectively. The seroprevalence of SIV H1N1 according to herd size was not significant differences (p>0.05). The results showed that the PRRSV infection spread widely in swine herds throughout the country.
Apple stem grooving virus (ASGV) is a high-risk viral pathogen that infects many types of fruit trees, especially pear and apple, and causes serious economic losses across the globe. Thus, rapid and reliable detection assay is needed to identify ASGV infection and prevent its spread. A reliable reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed, optimize, and evaluated for the coding region of coat protein of ASGV in pear leaf. The developed RT-LAMP facilitated the simple screening of ASGV using visible fluorescence and electrophoresis. The optimized reaction conditions for the RT-LAMP were 63℃ for 50 min, and the results showed high specificity and 100-fold greater sensitivity than the reverse transcription polymerase chain reaction. In addition, the reliability of the RT-LAMP was validated using field-collected pear leaves. Furthermore, the potential application of paper-based RNA isolation, combined with RT-LAMP, was also evaluated for detecting ASGV from field-collected samples. These assays could be widely applied to ASGV detection in field conditions and to virus-free certification programs.
Capsicum annuum (CA) is grown outdoors across fields in Jeollabuk-do, South Korea. The weeds surrounding these fields were investigated regarding the infection of 11 viruses infecting CA during the year 2014-2018. In the reverse transcription polymerase chain reaction diagnosis, 546 out of 821 CA samples (66.5%) were infected by nine viruses, and 190 out of 918 weed samples (20.7%) were infected by eight viruses. Correlation analysis of the mutual influence of the viruses infecting CA and weeds during these 5 years showed that five viruses had significant positive correlations with the infection in both CA and weeds. Over the study period, the weeds infected by cucumber mosaic virus (CMV) in the previous year were positively correlated with the incidence of CMV infection in CA in the current year, although the correlation was lower for tomato spotted wilt virus (TSWV) compared to CMV. The CMV infection percent was 14.0% in summer annuals, 11.4% in perennials, and 7.8% in winter annuals. However, considering the overwintering period without CA, the infection percent was 5.2% higher in winter annuals and perennials than that in summer annuals, indicating that winter annual and perennial weeds served as the main habitats for insect vectors. The TSWV infection percent in weeds was 10.4% in summer annuals, 6.4% in winter annuals, and 6.2% in perennials. The weeds surrounding CA fields, acting as the intermediate hosts, were found to be the potent sources of infection, influencing the spread and diversity of CA-infecting viruses. The results of this study can contribute to prevent viral infection in agricultural fields.
The global spread of flaviviruses has triggered major outbreaks worldwide, significantly impacting public health, society, and economies. This has intensified research efforts to understand how flaviviruses interact with their hosts and manipulate the immune system, underscoring the need for advanced research tools. RNA-sequencing (RNA-seq) technologies have revolutionized our understanding of flavivirus infections by offering transcriptome analysis to dissect the intricate dynamics of virus-host interactions. Bulk RNA-seq provides a macroscopic overview of gene expression changes in virus-infected cells, offering insights into infection mechanisms and host responses at the molecular level. Single-cell RNA sequencing (scRNA-seq) provides unprecedented resolution by analyzing individual infected cells, revealing remarkable cellular heterogeneity within the host response. A particularly innovative advancement, virus-inclusive single-cell RNA sequencing (viscRNA-seq), addresses the challenges posed by non-polyadenylated flavivirus genomes, unveiling intricate details of virus-host interactions. In this review, we discuss the contributions of bulk RNA-seq, scRNA-seq, and viscRNA-seq to the field, exploring their implications in cell line experiments and studies on patients infected with various flavivirus species. Comprehensive transcriptome analyses from RNA-seq technologies are pivotal in accelerating the development of effective diagnostics and therapeutics, paving the way for innovative treatments and enhancing our preparedness for future outbreaks.
Zhang, Yubao;Xie, Zhongkui;Fletcher, John D;Wang, Yajun;Wang, Ruoyu;Guo, Zhihong;He, Yuhui
The Plant Pathology Journal
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v.36
no.1
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pp.76-86
/
2020
Cucumber mosaic virus (CMV) is damaging to the growth and quality of lettuce crops in Lanzhou, China. Recently, however, for the first time an isolate of lettuce necrotic yellows virus (LNYV) has been detected in lettuce crops in China, and there is concern that this virus may also pose a threat to lettuce production in China. Consequently, there is a need to develop a rapid and efficient detection method to accurately identify LNYV and CMV infections and help limit their spread. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays were developed to detect the nucleoprotein (N) and coat protein (CP) genes of LNYV and CMV, respectively. RT-LAMP amplification products were visually assessed in reaction tubes separately using green fluorescence and gel electrophoresis. The assays successfully detected both viruses in infected plants without cross reactivity recorded from either CMV or LNYV or four other related plant viruses. Optimum LAMP reactions were conducted in betaine-free media with 6 mM Mg2+ at 65℃ for LNYV and 60℃ for 60 min for CMV, respectively. The detection limit was 3.5 pg/ml and 20 fg/ml using RT-LAMP for LNYV and CMV plasmids, respectively. Detection sensitivity for both RT-LAMP assays was greater by a factor of 100 compared to the conventional reverse transcription polymerase chain reaction assays. This rapid, specific, and sensitive technique should be more widely applied due to its low cost and minimal equipment requirements.
In this study, the virucidal efficacy of a fumigant containing 20% ortho-phenylphenol against classical swine fever virus (CSFV) and porcine reproductive and respiratory syndrome virus (PRRSV) was examined. After each carrier deposited with CSFV and PRRSV suspensions was exposed to the fumigant in a $25-m^3$ test room for 15 h, all carriers were neutralized and diluted, and each diluted suspension was inoculated into each proper cell line. After incubation, CSFV and PRRSV viability in each cell line was examined and 50% tissue culture infectious dose $(TCID_{50})/mL$ was calculated. In the results, the concentration of viable virus in all of pathogen control-carriers was more than $2{\times}10^5TCID_{50}/mL$, and there were no cytotoxicity in all of toxicity control-carriers. In addition, the fumigant inactivated ${\geq}4.8{\log}_{10}(TCID_{50}/mL)$ of both CSFV and PRRSV. These findings will be useful for preventing the spread of CSFV and PRRSV infection.
The foot and mouth disease and AI were highly contagious. The virus can be transmitted in a number of ways, including close-contact animal to animal spread, long-distance aerosol spread and fomites, or inanimate objects, typically fodder and motor vehicles. A lot of burial sites were constructed in a short time for preventing the rapid spread of the virus. The carcass burial sites have a risk potential because the sites were constructed without any appropriate and systematic management. It resulted from lacking of time, equipments and man power. The carcass burial sites more than 4,700 constructed in 2011. Approximately 7 million poultry and 3.5 million livestock including head of cattle and swine were buried in farm land. It is time to be concerned if the secondary pollutions occur from the burial sites. The environmental impacts should be analyzed for managing the burial sites effectively and minimizing damages and risks to the environment and human health. This study was to analyze environmental impacts of the process of carcass burial construction using a life cycle assessment methodology. All input data of raw materials and energy usage were collected and the inventory was constructed. The results showed that 1 ton of carcass burial of the environmental impacts were $0.51yr^{-1}$ for ADP, 0.09 kg of 1,4DCB-eq for FAETP, 31.17 kg of $CO_2-eq$ for GWP, 0.04 kg of $C_2H_4-eq$ for POCP, 0.06 kg of $SO_2-eq$ for AP.
In March 2003, classical swine fever (CSF) infection was reported in a piggery located at Iksan city, Jeollabuk-do in Korea. Subsequently, a total of 72 infected farms were confirmed between March and December, 2003. Based on epidemiological investigation of the earlier confirmed infected farms, the source of infection was shown to be from a breeding farm. Targeted surveillance of 82 piggeries that had acquired pigs from this breeding farm showed 44 piggeries were infected with CSF virus. CSF virus was introduced into this breeding farm by movement of selected breeder pigs from its 12 contracted farms which were located in areas that had been affected by CSF epidemic in late 2002. CSF had then spread through out the country mainly by direct transmission through the sale and movement of pigs from this breeding farm. Consequently, 47 (62%) among 72 CSF affected farms were associated, directly and indirectly, with this breeding farm. This study showed that inadequate control for breeding farms and transport restriction in CSF outbreak areas resulted in the nationwide spread of CSF and the failure of the eradication campaign that has been underway for several years by the Korean animal hygiene authority as well as the fanners. Improvements of control policy through further research of the 2003 CSF epidemic will be needed to reestablish the Korean CSF eradication program in the future.
Isolates of soybean mosaic virus (SMV) strains classified based on virulence in silt resistant soybean cultivars caused the same reactions in soybean cultivars used as differentials as those obtained by sap inoculations to the same cultivars. Five species of aphids (Myzus persicae SULZ., Aphis craccivora KOCH, Aphis citricola VAN., Rhopalosiphum maidis FIT., End R. padi L.) were able to transmit each of SMV strains. However, R. maidis and R. padi were inefficient vectors for transmission of SMV strain G3. Spread if four SMV strains (G2, G3, G6, and G7) was monitored in the field from sapinoculated plants in a one meter row of Williams soybeans (source plants) to plants in an adjacent row of Williams 80cm away (test plants). Test plants wert downwind from the source plants. A complete block design was used. Spread of strain G6 was significantly greater than that of other three strains. Two hundred six aphids were collected from June 27, 1979 to August 2, 1979 in the same field. A. citricola was the mist prevalent, comprising $68\%$ of the total aphids. Yields of Williams inoculated with each strain were also compared. Yields were the least from plants inoculated with strain G2 following G6, G3, and G7 in that order.
Kim, Jung-Mi;Song, Ha-Yeon;Yun, Suk-Hyun;Lee, Hyun-Suk;Ko, Han-Kyu;Kim, Dae-Hyuk
한국균학회소식:학술대회논문집
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2015.11a
/
pp.37-37
/
2015
dsRNA was found in malformed cultures of Lentinula edodes strain FMRI0339, one of the three most popular sawdust cultivated commercial strains of shiitake, and was also found in healthy-looking fruiting bodies and actively growing mycelia. Cloning of the partial genome of the dsRNA revealed the presence of the RdRp sequence of a novel L. edodes mycovirus (LeV), and sequence comparison of the cloned amplicon showed an identical sequence to known RdRp genes of LeV found in strain HKA. The meiotic stability of dsRNA was examined by measuring the ratio of the presence of dsRNA among sexual monokaryotic progeny. More than 40% of the monokaryotic progeny still contained the dsRNA, indicating the persistence of dsRNA during sexual reproduction. Comparing the mycelia growth of monokaryotic progeny suggested that, although variations in the growth rate existed among progeny and virus infection was observed in highly actively growing progeny, there appeared to be a tendency toward a lower frequency of virus incidence in actively growing progeny. This study attempted to cure the edible mushroom L. edodes strain FMRI0339 of the L. edodes mycovirus (LeV) in order to obtain an isogenic virus-free fungal strain as well as a virus-infected strain for comparison. Mycelial fragmentation, followed by being spread on a plate with serial dilutions resulted in a virus-free colony. Viral absence was confirmed with gel electrophoresis after dsRNA-specific virus purification, Northern blot analysis, and PCR using reverse transcriptase (RT-PCR). Once cured, all of fungal cultures remained virus-free over the next two years. Interestingly, the viral titer of LeV varied depending on the culture condition. The titer from the plate culture showed at least a 20-fold higher concentration than that grown in the liquid culture. However, the reduced virus titer in the liquid culture was recovered by transferring the mycelia to a plate containing the same medium. In addition, oxygen-depleted culture conditions resulted in a significant decrease of viral concentration, but not to the extent seen in the submerged liquid culture. Although no $discernable phenotypic changes in colony morphology were observed, virus-cured strains showed significantly higher growth rates and mycelial mass than virus-infected strains. We were also explored effects of LeV on fruiting body formation and mushroom yield. The fruiting body formation yield of virus-free L. edodes was larger than virus-infected L. edodes. These results indicate that LeV infection has a deleterious effect on mycelial growth and fruiting body formation. In addition, we have been investigated host-parasite interaction between L. edodes and its mycovirus interaction to study viral mechanism by establishment of proteomics.
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