To identify viruses and compare their abundance levels in the venom glands of hymenopteran species, we conducted venom gland-specific transcriptome assemblies and analyses of 22 Aculeate bees and wasps and identified the RNA genomes of picornaviruses. Additionally, we investigated the expression patterns of viruses in the venom glands over time following capture. Honeybee-infecting viruses, including black queen cell virus (BQCV), deformed wing virus (DWV), and Israeli acute paralysis virus (IAPV), were highly expressed in the venom glands of Apis mellifera and social wasps. This finding suggests that the venoms of bees and wasps likely contain these viruses, which can be transmitted horizontally between species through their stinger usage. A. mellifera exhibited an increasing pattern of abundance levels for BQCV, DWV, IAPV, and Triatovirus, while the social wasp Vespa crabro showed increasing abundance levels of IAPV and Triatovirus over different capture periods. This suggests that the venom glands of honeybees and wasps may provide suitable conditions for active viral replication and may be an organ for virus accumulation and transmission. Some viral sequences clearly reflected the phylogeny of Aculeate species, implying host-specific virus evolution. On the other hand, other viruses exhibited unique evolutionary patterns of phylogeny, possibly caused by specific ecological interactions. Our study provides insights into the composition and evolutionary properties of viral genes in the venom glands of certain Aculeate bees and wasps, as well as the potential horizontal transmission of these viruses among bee and wasp species.

South Korea experienced a significant decline in honey bee populations starting in 2021, which continued for two years until the winter of 2022. To investigate the potential causes of this decline, we conducted a virome analysis, considering viruses as possible culprits. Samples were collected during two periods: April-May 2022 and May-June 2023. From libraries contsructed from their total RNA, we secured a total of 25 raw FASTQ files by high-throughput sequencing. In the honey bees collected in 2022, we identified eight previously unreported honey bee viruses including Lake Sinai viruses, one novel honey bee-related virus, and one novel plant-related virus. In the subsequent sampling in 2023, we found that most of the viruses identified in 2022 were still present. Additionally, the novel honey bee virus reported in 2022 was also found in the 2023 collections, along with three more honey bee-related novel viruses. Notably, numerous plant viruses were detected in honey bees collected during the flowering season. This analysis suggests that the viruses observed in South Korean honey bees are likely distributed nationwide. These findings provide fundamental data for future research on honey bee viruses in South Korea.

Over the course of two winters, the significant decline in honey bee populations in Korea has emerged as a major social issue. This phenomenon is expected as attributed to factors such as the failure of pest control due to the pesticide resistance of the Varroa mite. This mite can transmit some viruses that infect honey bees, and these viruses are among the primary causes of the globally occurring colony collapse disorder. Traditional diagnostic methods like (RT-)PCR and ELISA are not ideal for identifying pathogens that are newly emerging or have undergone mutations. To detect any novel or mutated viruses beyond those that have been primarily diagnosed in Korea, we introduced virome analysis technology in the field of honey bees. Employing this method with high-throughput sequencing techniques, we were able to identify all existing viruses within individual or group samples. We discovered that the Lake Sinai virus, which has been reported worldwide but not in Korea, has already significantly spread within the country. Additionally, we were able to confirm the prevalence of viruses previously reported in Korea, such as the recently dominant Black Queen Cell Virus. Through this virome analysis, we can provide foundational data for determining the direction and countermeasures for virus diagnosis.

The red imported fire ant (Solenopsis invicta) is a species of ant native to South America. The fire ant was inadvertently introduced into USA, Australia, New Zealand, and other Asian countries including China and Taiwan. Since the first report of the fire ant in port city of Busan, Korea in 2017, it was found in many other cities of Korea in following year. To obtain the molecular information of this invasive species, total RNA was extracted from the abdominal segment of the ants collected in Incheon, and subjected to transcriptome sequencing. By using Illumina sequencer platform, 101 base pared-end sequencing generated 2 × 50,064,081 of raw reads to obtain 2 × 45.95 Gbase of quality filtered nucleotide sequences. The in silico cDNA library was constructed by Trinity de novo assembler followed by TransDecoder ORF finder and CD-HIT clustering program to streamline the library. The final version of cDNA library contains 20,442 contigs with protein coding capability. To survey the virome of this ant, these contigs were searched against the viral reference sequences from NCBI RefSeq database with BLASTN program. As a result, contigs which showed high sequence identities with several RNA viruses including previously reported SINV-2 were found from the fire ant. This virome information might give an idea of a shift of virological environment of this newly found ant isolate or population in Korea.

Frequent surveys and monitoring were conducted in the Southern part of Bangladesh to detect and identify the plant pathogenic virus that infecting agriculturally important vegetables during 2017-2018. A total of 28 fields of the survey area were closely monitored. The findings indicated that 21.94% of the plants developed typical virus disease like symptomps in the field. However, 28.21% infected plants were found in Patuakhali followed by Satkhira (23.11%), Khulna (19.33%) and Barguna (17.12%). The symptoms were mosaic, vein clearing, chlorosis, curling and ringspot. Twenty samples from the collections were randomly chosen on the basis of symptoms and subjected to Enzyme-linked Immunosorbent Assay (ELISA) with the antiserum and symptomalogy were used for detection. Eight viruses namely Cucumber mosaic virus (CMV), Okra yellow vein clearing mosaic virus (OYVCMV), Mungbean yellow mosaic virus (MYMV), Tomato yellow leaf curl virus (TYLCV), Pepper mottle virus (PMV), Papaya ringspot virus (PRSV), Watermelon mosaic virus (WMV) and Zucchini yellow mosaic virus (ZYMV) were detected on cucumber, okra, brinjal, mungbean, tomato, pepper, papaya, watermelon and pumpkin respectively.

The varroa mite, Varroa destructor, is a small ectoparasitic mite which attacks honeybee, Apis mellifera, and also known to harbor small RNA viruses which infect honeybees. To survey the transcriptome of varroa mite, total RNA of female adult mites was subjected to RNA-seq to construct an in silico cDNA library. 2 × 8.3 Gbase of quality filtered paired-end nucleotide sequences were obtained to construct 28,302 of protein-coding contigs by de novo assembly, and subsequent BLAST search revealed the viruses infect honeybee or associated with varroa mites. Six of the contigs showed high sequence identity to Iflavirus, picorna-like virus, rhabdovirus, and macula-like virus were discovered. It implies that the viral flora in varroa mites and honeybees might be more complex than previously studied, and suggests the importance of further virome studies for better understanding of honeybee health.

Plant virus can enhance its transmission by altering the settling preference of its vector. Nevertheless, most of the studies have focused on the spread of one virus in a field whereas often times there are more than single virus infecting same crop. Furthermore, mixed-infection of multiple viruses tends to cause more severe virus symptoms and changes vector’s biology and behavior different than singular infection. Thus, we are currently investigating the spatial transmission pattern of persistently transmitted potato leafroll virus and non-persistently transmitted potato virus y. However, due to impracticability of obtaining empirical data, we are programming an individual-based modelling software while taking biology of potato, biology and behavior of aphid and different characteristics of two viruses into consideration.

종자의 수입 시, 검역관련 종자전염바이러스는 가장 문제가 되는 식물병이다. 본 연구에서 PCR 검역체계가 보고되지 않은 3종의 종자전염바이러스, Cherry rasp leaf virus (CRLV), Spinach latent virus (SpLV) 및 White clover mosaic virus (WClMV)를 검출하기 위하여 reverse transcription polymerase chain reaction (RT-PCR)과 nested polymerase chain reaction (nested PCR) 방법을 도입하였다. 각각의 바이러스별로 2 세트의 RT-PCR primer가 선발되었으며, 증폭산물에서 더욱 높은 감도로 검출 할 수 있는 nested PCR primer set를 개발하였다. 본 연구에서 사용한 RT-PCR과 nested PCR 방법은 종자로부터 CRLV, SpLV 및 WClMV를 검역하는 고효율적 진단시스템으로 제공될 것이다.

바이러스 입자를 감지하는 역전사 핵산 연쇄 증폭법 (VC/RT-PCR)은 감염된 식물종들로부터 핵산 추출 없이 식물바이러스들을 검출 할 수 있다. 본 연구는 VC/RT-PCR 분석법을이용하여 고추를 감염시키는 바이러스들을 효과적으로 진단하기 위하여 새로운 즙액 추출 완충액들이 제작하였다.토마토반점위조바이러스 (Tomato spotted wilt virus;TSWV), 고추약한모틀바이러스 (Pepper mild mottle virus;PMMoV) 및 고추모틀바이러스 (Pepper mottle virus;PepMoV) 진단을 위한 가장 최적화된 추출 완충액은 0.5%sodium sulfate를 포함하는 1.0M Tris (pH 8.0) buffer 였다.고추 바이러스들은 담배 즙액 추출 후 7일까지 검출이 되었으며, 마쇄 직후와 검출 감도는 유의한 차이가 없었다. 반면에,3가지 고추 바이러스들은 고추 즙액 추출 후 2일까지만 바이러스들이 검출되었으며, 검출 감도는 크게 감소하였다.국내 고추 재배 농가들에서 수집한 고추 시료들에서 TSWV,PMMoV, PepMoV의 단독 감염 및 PMMoV와 PepMoV의중복 감염을 선발된 최적 즙액 완충액과 VC/RT-PCR의 조합을 이용하여 동시 진단이 가능하였다.

오염된 식품으로부터 바이러스를 효과적으로 검출하기 위해서는 식품에 부착된 바이러스를 효과적으로 elute시키는 것이 결정적으로 중요하다. 본 연구에서는 채소류에 공통적으로 적용할 수 있는 elution용액을 찾기 위해 폴리오 바이러스를 인위적으로 오염시킨 1가지 엽채류 (깻잎) 와 3가지 근채류 (당근, 양파, 무) 로부터 바이러스 회수율을 조사한 후, 최적의 바이러스 회수조건을 찾기 위해 기보고된 상추와 양배추의 회수율을 함께 분석하였다. 바이러스의 회수율은 식품의 matrix와 사용된 elution용액의 종류에 따라 차이가 컸으나 0.25M threonine / 0.3M NaCl (pH 9.5) 또는 0.25M glycine / 0.14M NaCl (pH 9.5)을 사용하였을 때 6가지 채소 중 5가지로부터 폴리오바이러스를 효과적으로 elute 할 수 있었다. 0.25M threonine / 0.3M NaCl (pH 9.5)를 노로바이러스 검출에 적용해 본 결과 근채류인 당근보다 엽채류인 깻잎으로부터 노로바이러스 GII를 더 잘 검출할 수 있었다. 이 같은 공통 elution용액을 사용할 경우 다양한 종류의 채소류에 오염된 노로바이러스를 포함한 소화기바이러스의 검출을 용이하게 해 줄 것이다.

In the transcriptome surveys of Laodelphax striatellus, several cDNA sequences showed a high level of similarities to the insect picorna-like virus genomes. Interestingly, there was no sequence similarity between picorna-like virus sequences from the RSV-viruliferous and those from the non-viruliferous L. striatellus. Picorna-like virus from the non-viruliferous L. striatellus was a geographical isolate of Himetobi P virus (HiPV). The genome of the HiPV was 9,272 nt in length excluding the poly(A) tail and contained two open reading frames (ORFs), which were separated by a 176 nt intergenic region that functions as an internal ribosome entry site (IRES). The 5' ORF encodes the non-structural proteins and the 3' ORF encodes the capsid proteins. The partial genomic RNA of the picorna-like virus from the RSV-viruliferous L. striatellus, LsPV-2, was 8,769 nt in length excluding the poly(A) tail and contained a single, large open reading frame (nt 1–8,535) encoding a 2,845 aa polyprotein. In terms of sequence similarity, identity, and genome organization, LsPV-2 resembled insect picornalike viruses belonging to the family Iflaviridae. A phylogenetic analysis based on RNA-dependent RNA polymerase (RdRp) sequence showed that LsPV-2 was most closely related to the deformed wing virus (DWV). The HiPV and LsPV-2 were incompatible each other in L. striatellus, suggesting that these two picorna-like viruses may have important functions in transmission of the RSV.

Sperm examination is an important tool in estimating the fertilizing capacity of an ejaculate. The number of spermatozoa in a semen dose, morphology and motility are important for the fertilization process. By evaluation of semen, artificial insemination (AI) using high quality of semen can increase fertilization rate. Boar semen is subject to contamination by various pathogens that can result in fertility disorders in sows. Among these pathogens, porcine reproductive and respiratory syndrome virus (PRRSV), porcine parvovirus (PPV), porcine circovirus-2 (PCV-2) are of particular importance and accurate monitoring prior to and during the presence of boars in AI stations is essential. Because of the high risk of dissemination of disease via AI, The absolute goal is to provide pathogen-free semen and this is feasible with the adequate measures. The disease affects boars semen causes a significant reduction quality. In this study we investigated the characterization boar semen in Jeju, interaction of pathogenic virus infection with characterization of boar semen. Forty-two boar semen from 13 farms were investigated. The semen were stored during 5 days at 17℃ and the sperm qualities in the stored semen were analysed. Visual-motility assessment is a tool (Computer- Assisted Semen Analysis) used to determine the quality of boar semen. Percentage of morphologically normal spermatozoa were assessed. PRRS ,PPV and PCV-2 were detected in boar semen using PCR. The motion characteristics in boar semen was showed 68.4±9.1% for motility, 48.6±7.1 μm/s for VAP, 45.3±7.0 μm/s for VSL, 79.1±8.7 μm/s for VCL, 1.3±0.2 μm/s for ALH, 8.3±0.4 Hz for BCF, 93.6±3.5% for STR, 57.9±6.4 % for LIN. The percentage of sperm with abnormal head, midepeice and tail were 0.3±0.7%, 14.4±12.5%, 4.9±6.6%, respectively. Based on the PCR method, PPV was detected in 20 samples (48%). However, PCV-2 and PRRSV were not detected in any cases. Marked differences in motility and morphology between PPV negative and PPV positive semen were not observed. Sperm cell production was not affected by PPV infection. However, slight increases in detached head, coiled tail after infection were observed (p<0.05). The motility of semen in Jeju is similar to case comparing with other regions in Korea. Although PPV in semen was not affected in semen quality, there is the high risk of virus excretion in the semen of Jeju boars. Therefore continuous screening tests for some particular pathogens in boar semen would be warranted.

Thailand suffers from one of the highest rates of dengue and dengue haemorrhagic fever in the world. Since there is no vaccine or any specific treatments for dengue, currently, the only effective way to prevent dengue fever and dengue hemorrhagic fever is vector control. The knowledge of basic biology, ecology, behavior and survival factors of Ae. aegypti is crucial to control mosquitoes. The impact of the environmental factors; temperature, rainfall, and humidity, on the ecology, biology, development, vector density, behavior, fitness of mosquito vectors, and the mosquito-borne disease transmission dynamics will be presented and discussed.