Haemaphysalis longicornis는 사람과 동물에게 여러 심각한 병원체를 전달하는 주요 매개체로, 한반도에 널리 분포하고 있다. H. longicornis는 Rickettsia spp., Borrelia spp., Francisella spp., Coxiella spp., 그리고 중증열성혈소판 감소증후군 바이러스 (SFTS virus) 등을 매개하는 것으로 알려져 있다. 국내에 서식하는 H. longicornis의 미생물 군집과 관련된 연구는 많이 진행되지 않은 것으로 확인되었다. 이 연구는 한반도 내 다양한 지역에서 채집된 H. longicornis의 미생물군집 다양성을 지역별, 성장 단계 및 성별에 따라 분석하였다. 2019년 6월부터 7월까지 질병관리청 권역별기후변화매개체감시거점센터 16개 지역에서 채집한 H. longicornis의 16S rRNA 유전자 V3-V4 영역을 PCR로 증폭 후 Illumina MiSeq 플랫폼으로 시퀀싱하였다. Qiime2를 활용한 미생물 다양성 분석을 통해 총 46개의 샘플에서 1,754,418개의 non-chimeric reads를 얻었으며, 평균 126개 의 operating taxonmic unit (OTU) 을 식별하여 총 1,398개의 OTU를 확인하였다. 대부분의 지역에서 Coxiella spp.가 우점종으로 나타났으며, 특히 Coxiella endosymbiont는 가장 높은 우점도를 보이며, Coxiella burnetii와 계통 발생 학적으로 유사한 것으로 확인되었다. 이 연구를 통해 분석된 결과는 각 지역의 H. longicornis 미생물군집 데이터 베이스 구축에 활용되었으며, 이를 통해 지역별 미생물군집의 특이성을 식별할 수 있게 하였다. 이는 한반도의 H. longicornis에 의한 질병 전파 연구와 이를 통한 공중보건 개선에 기여할 것으로 기대된다.
Recently, it is demonstrate that the invertebrates have a immune memory, called Immune priming (IP). It was partially studied that the IP is mainly regulated by epigenetic modification. Here, to understand the IP on antimicrobial peptides (AMPs) production, we investigated larval mortality and time-dependent expression patterns of AMP genes in T. molitor larvae challenged with E. coli (two-times injection with a one-month interval). Interestingly, the results indicate that the higher and faster expression levels of most AMP genes were detected compared to the non-primed T. molitor larvae. Our results may used to improve the understanding of mechanisms of invertebrate immune memory.
Pellino, a highly conserved E3 ubiquitin ligase, is known to mediate ubiquitination of phosphorylated Interleukin-1 receptor-related kinase (IRAK) homologs in Toll signaling pathway. To understand the immunological function of TmPellino, we screened the knockdown efficiency of TmPellino by injecting TmPellino-specific dsRNA into T. molitor larvae. Subsequently, we investigated the larval mortality and the tissue-specific expression patterns of antimicrobial peptide (AMP) genes against microbial challenges. Interestingly, the results indicate that the expression of many AMP genes was upregulated in the Malpighian tubules of TmPellino-silenced T. molitor larvae. This study may provide basic information to understand how Tmpellino regulates AMPs production in T. molitor.
Tumor necrosis factor receptor-associated factor (TRAF) is known to regulate antimicrobial peptides (AMPs) production in mammals. Here, to understand the immunological function of TmTRAF against microbial challenge, the induction patterns of TmTRAF against microbial infection was investigated by qRT-PCR in the whole-body and tissue of young larvae. In addition, the effects of TmTRAF RNAi on larval mortality and expression of 15 AMP genes in response to microbial infection were investigated. Our studies may help to understand the basic role of AMP production.
Tube, an intracellular protein of the Toll-pathway, forms a complex with Pelle and MyD88, and regulates a signal transduction to activate NF-κB in Drosophila. To understand the antimicrobial function of TmTube, the induction patterns of TmTube were investigated at 3, 6, 9, 12, and 24 h-post injection of pathogens into 10th to 12th instar larvae. In addition, we investigated the effects of TmTube RNAi on larval mortality and tissue specific AMP expression in response to microbial challenge. Our results will provide a basic information to elucidate the immunological function of TmTube
Pelle, a serine/threonine kinase, is an intracellular component of the Toll pathway and is involved in antimicrobial peptides (AMPs) production due to pathogenic infection. It is known that the Pelle phosphorylates Cactus and activates the NF-κB signaling pathway in Drosophila, but it is not studied in Tenebrio molitor. In this study we investigated the tissue-specific expression patterns of the Pelle following pathogenic infection at 3, 6, 9, 12, and 24 hours. Additionally, larval mortality and AMP expression against microbial injection were investigated in dsPelle-treated T. molitor larvae. Our results may help to understand the antimicrobial function of TmPelle.
In insects, the glutathione S-transferase is initiated in both the detoxification process and the protection of cellular membranes against oxidative damage. In this study, we identified the open reading frame (ORF) sequence of GST-iso1 and 2 from Tenebrio molitor (TmGST-iso1 and 2). To investigate the expression patterrns of TmGST-iso1 and 2 in response to herbicide, 0.06, 0.6, and 6 ㎍/㎕ of butachlor (FarmHannong, Seoul, South Korea) was challenged into T. molitor larvae, resulting that the TmGST-iso1 were highly induced at 3 and 24 h-post injection. Whereas, the highest expression of TmGST-iso2 was detected at 24 h after treatment. This study may contribute to basic information about the detoxifying activities of T. molitor.
It is well known that the JNK pathway regulates AMP production against pathogenic infection in both vertebrates and invertebrates. Tenebrio molitor hep (Tmhep) is an homolog of MAP kinase kinase in mammals. Here, we investigate the immunological function of Tmhep in responses in microbial infection using RNA interference technology. The results showed that silencing of Tmhep increased the larval mortality against microbial challenge, as well as reduced AMP production compared to the control group (dsEGFP-treated group). Conclusively, Tmhep plays an critical role in antimicrobial defense in T. molitor larvae.