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During December 2012-July 2016, we tested small indoor and outdoor mammals in Qingdao, China, for Orientia tsutsugamushi infection. We found that outdoor Apodemus agrarius mice, Cricetulus barabensis hamsters, and Niviventer confucianus rats, as well as indoor Mus musculus mice, tested positive for O. tsutsugamushi by PCR.

Microsporidia are a group of unicellular and opportunistic intestinal parasites in which Enterocytozoon bieneusi is a frequent species causing microsporidial infections in humans. Many domesticated and wild animals have been shown to be hosts of E. bieneusi and other microsporidia. The role of hedgehogs in the ecology of microsporidia is unclear; therefore, we investigated the prevalence and genetic diversity of E. bieneusi, Cryptosporidium, and Blastocystis spp. in hedgehogs (Erinaceus amurensis) collected from Hubei Province in Central China. PCR amplification of the internal transcribed spacer region of the ribosomal DNA indicated that 9.8% (4/41) hedgehogs were positive to E. bieneusi, but none (0/41) was positive to Cryptosporidium and Blastocystis spp. Phylogenetic analysis showed the strains detected from the hedgehogs belong to four novel genotypes (EA1-EA4), which were most closely related to type IV of group 1c. This study demonstrated that hedgehogs are hosts of E. bieneusi and may play a role in the transmission of E. bieneusi to humans in the process of being caught and slaughtered.

Coxiella burnetii is the causative agent of query fever (Q fever), and distributes broadly in environment. Livestock are identified as main reservoirs, which may infect people through their contaminative urine, feces, milk, and birth products. Wild animals can also be the potential carriers and transmitters of C. burnetii. To understand the geographic distribution and host species of C. burnetii in China, we investigated the prevalence of C. burnetii in hedgehogs (Erinaceus amurensis) in Hubei Province. Hedgehogs were tested for C. burnetii with PCR targeting three genes (com1, rrs, and icd) followed by multispacer sequence typing (MST). We found that 12.2% (5/41) hedgehogs were PCR positive for C. burnetii. MST revealed presence of two novel genotypes and phylogenetic analysis revealed that the strains were similar to a group of isolates from chronic Q fever patients and mammals. This study showed that C. burnetii are highly prevalent in hedgehogs in Hubei Province in central China, suggesting that hedgehogs may play an important role in the ecology and transmission of C. burnetii to humans because it is captured and used as traditional medicine in China.

Background: Leptospira is the causative agent of leptospirosis, a zoonotic disease of global importance. To have a better understanding on the host species of Leptospira, we investigated the preval...

Snakes are popular as food and traditional medicine in China. However, information about parasitic and bacterial infections in snakes from China is scarce. We investigated the prevalence of selected zoonotic agents including Cryptosporidium, Hepatozoon and Spirometra, in snakes in central China from June to October in 2018 by PCR amplification using parasite-specific primers. PCR amplification and DNA sequencing showed that 10.1% (15/149) of snakes were positive for Cryptosporidium spp., while 2.7% (4/149) were positive for Hepatozoon. Additionally, we found 36.9% (55/149) of snakes were infected with Spirometra erinaceieuropaei. The spargana burden per infected snake ranged from 1 to 26. BLAST and phylogenetic analysis of small subunit ribosomal RNA (SSU rRNA) gene and 60-kDa glycoprotein (gp60) gene showed that the parasites belonged to Cryptosporidium parvum genotype IIdA15G1, C. baileyi, C. serpentis and a Hepatozoon species. We conclude that intensively farmed snakes excrete C. parvum and C. baileyi oocysts due to ingestion of infected feeder animals, and that wild snakes in central China were commonly infected with S. erinaceieuropaei, suggesting that eating improperly cooked snakes could be risky to human health., Graphical abstract Image 1, Highlights • Cryptosporidium parvum and C. baileyi were reported in farmed snakes in China for the first time. • C. serpentis was detected for the first time in Ptyas dhumnades and Gloydius brevicaudus. • A tentative new Hapatozoon species was first time recorded in wild Lycodon rufozonatus and G. brevicaudus. • Spirometra erinaceieuropaei infection was found common in wild snakes from central China.

Nuclear scaffold attachment factor A (SAFA) is a novel RNA sensor involved in sensing viral RNA in the nucleus and mediating antiviral immunity. Severe fever with thrombocytopenia syndrome virus (SFTSV) is a bunyavirus that causes SFTS with a high fatality rate of up to 30%. It remains elusive whether and how cytoplasmic SFTSV can be sensed by the RNA sensor SAFA. Here, we demonstrated that SAFA was able to detect SFTSV infection and mediate antiviral interferon and inflammatory responses. Transcription and expression levels of SAFA were strikingly upregulated under SFTSV infection. SAFA was retained in the cytoplasm by interaction with SFTSV nucleocapsid protein (NP). Importantly, SFTSV genomic RNA was recognized by cytoplasmic SAFA, which recruited and promoted activation of the STING-TBK1 signaling axis against SFTSV infection. Of note, the nuclear localization signal (NLS) domain of SAFA was important for interaction with SFTSV NP and recognition of SFTSV RNA in the cytoplasm. In conclusion, our study reveals a novel antiviral mechanism in which SAFA functions as a novel cytoplasmic RNA sensor that directly recognizes RNA virus SFTSV and mediates an antiviral response., Author summary Severe fever with thrombocytopenia syndrome virus (SFTSV) is an RNA virus with a high fatality rate of up to 30%, which replicates exclusively in the cytoplasm. To date, many cytoplasmic RNA sensors were known to recognize SFTSV infection and trigger antiviral immune responses. Nuclear scaffold attachment factor A (SAFA) is a novel nuclear RNA sensor which can sense viral RNA in the nucleus and promote activation of antiviral immunity. However, there are no studies to investigate whether SAFA could detect cytoplasmic RNA virus infection. Here, we reported that SAFA was able to detect RNA virus SFTSV invasion. Under SFTSV infection, SAFA was retained in the cytoplasm and recognized SFTSV infection by interaction with SFTSV nucleocapsid protein (NP) and cytoplasmic SFTSV RNA directly. Importantly, SAFA recruited and promoted the activation of the STING-TBK1 signaling pathway-mediated antiviral immunity to suppress SFTSV infection. This study provides a further acquaintance in SAFA-mediated antiviral immune responses, illustrating the novel role of SAFA in sensing cytoplasmic SFTSV and mediates an antiviral response.

Bin Ren,1,2,* Zhong-Chao Cai,1,* Xue-Jie Zhao,1 Lin-Song Li,1 Mei-Xia Zhao1 1Key Laboratory of Natural Medicine and Immuno-Engineering of Henan Province, Henan University, Jinming Campus, Kaifeng, Henan, 475004, People’s Republic of China; 2School of Mathematics and Statistics, Henan University, Jinming Campus, Kaifeng, 475004, People’s Republic of China*These authors contributed equally to this workCorrespondence: Mei-Xia ZhaoKey Laboratory of Natural Medicine and Immune Engineering of Henan Province, Henan University, Jinming Campus, Kaifeng, Henan, 475004, People’s Republic of ChinaEmail zhaomeixia2011@henu.edu.cnPurpose: Gold nanoparticles (AuNPs) with good physical and biological properties are often used in medicine, diagnostics, food, and similar industries. This paper explored an AuNPs drug delivery system that had good target selectivity for folate-receptor overexpressing cells to induce apoptosis.Methods: A novel drug delivery system, Au@MPA-PEG-FA-PTX, was developed carrying paclitaxel (PTX) on folic acid (FA) and polyethylene glycol (PEG)-modified AuNPs. The nanomaterial was characterized by transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy (FTIR), and ultraviolet-visible spectroscopy (UV-Vis). Also, the biological activity of the AuNPs drug delivery system was examined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in HL-7702, Hela, SMMC-7721, and HCT-116 cells. Furthermore, apoptotic activity using annexin V-FITC, mitochondrial membrane potential (MMP), and reactive oxygen species (ROS) levels was estimated by flow cytometry and fluorescence microscopy.Results: Au@MPA-PEG-FA-PTX exhibited a distinct core-shell structure with a controllable size of 28± 1 nm. Also, the AuNPs maintained good dispersion and spherical shape uniformity before and after modification. The MTT assay revealed good antitumor activity of the Au@MPA-PEG-FA-PTX against the Hela, SMMC-7721, and HCT-116 cells, while Au@MPA-PEG-FA-PTX produced better pharmacological effects than PTX in isolation. Further mechanistic investigation revealed that effective internalization of AuNPs by folate-receptor overexpressing cancer cells induced cell apoptosis through excessive production of intracellular ROS.Conclusion: The AuNPs drug delivery system showed good target selectivity for folate-receptor overexpressing cancer cells to induce target cell-specific apoptosis. These AuNPs may have great potential as theranostic agents such as in cancer.Keywords: gold nanoparticles, folic acid, paclitaxel, drug delivery system, apoptosis

Bartonella species are facultative intracellular bacteria and recognized worldwide as emerging zoonotic pathogens. Bartonella were isolated or identified by polymerase chain reaction (PCR) in bats and their ectoparasites worldwide, whereas the association between them was scarce, especially in Asia. In this study, a retrospective analysis with frozen samples was carried out to identify the genetic diversity of Bartonella in bats and their ectoparasites and to investigate the relationships of Bartonella carried by bats and their ectoparasites. Bats and their ectoparasites (bat flies and bat mites) were collected from caves in Hubei Province, Central China, from May 2018 to July 2020. Bartonella were screened by PCR amplification and sequencing of three genes (gltA, rpoB, and ftsZ). Bats, bat flies, and bat mites carried diverse novel Bartonella genotypes with a high prevalence. The sharing of some Bartonella genotypes between bats and bat flies or bat mites indicated a potential role of bat flies and bat mites as vectors of bartonellae, while the higher genetic diversity of Bartonella in bat flies than that in bats might be due to the vertical transmission of this bacterium in bat flies. Therefore, bat flies might also act as reservoirs of Bartonella. In addition, human-pathogenic B. mayotimonesis was identified in both bats and their ectoparasites, which expanded our knowledge on the geographic distribution of this bacterium and suggested a potential bat origin with bat flies and bat mites playing important roles in the maintenance and transmission of Bartonella.

Ipomoea aquatica, commonly known as water spinach, is an edible annual vegetable in the genus Ipomoea,. In this study, the complete plastome of Ipomoea aquatica was determined using the Illumina sequencing platform. The plastome size was 162,663 bp. It consists of four regions, including a large single-copy region (88,166 bp), a small single-copy region (12,069 bp), and a pair of inverted repeat regions (31,214 bp). This plastome encodes 114 unique genes, including 80 protein-coding genes (PCGs), 30 transfer RNA genes (tRNAs), and 4 ribosomal RNA genes (rRNAs). The GC content was 39.1%. Phylogenomic analysis based on 19 complete plastomes revealed that I. aquatica was closely related to I. diamantinensis.

We identified Candidatus Borrelia fainii, a human pathogenic bacterium causing New World relapsing fever in a Myotis bat in eastern China. This finding expands knowledge about the geographic distribution of Borrelia spp. and the potential for infection with New World relapsing fever in China.

Chemotherapy is currently a principal method for treating tumors. Selective delivery of chemotherapeutic drugs to the subcellular organs of cancer cells is an attractive treatment strategy for cancer. Mitochondria are important organelles that play complex roles in cancer, such as providing energy and controlling various cell death mechanisms. Specific induction of mitochondrial dysfunction can lead to irreversible cell death over a short duration, which is an effective method of destroying cancer cells. In this study, a novel nano-drug carrier (MSN-SS-CS-HA) for targeted drug delivery was prepared using mesoporous silica nanoparticles (MSNs), in which hyaluronic acid (HA) was used as a targeting ligand and blocking agent to control drug release. Coumarin is a fluorescent molecule that targets mitochondria. Two conjugates (XDS-DJ and 5-FUA-4C-XDS) were synthesized by chemically coupling nitrogen mustard and 5-fluorouracil with coumarin, which was further loaded into MSN-SS-CS-HA nanocarriers. MTT analysis demonstrated that the nanocomposite MSN-SS-CS@5-FUA-4C-XDS/HA displayed stronger cytotoxicity toward HCT-116 cells than HeLa or QSG-7701 cells. Furthermore, MSN-SS-CS@5-FUA-4C-XDS/HA was able to target the mitochondria of HCT-116 cells, causing decreased mitochondrial membrane potential and excessive production of reactive oxygen species. These results indicate that MSN-SS-CS@5-FUA-4C-XDS/HA has the potential as a nano-drug delivery system for the treatment of colon cancer.

Salvia hispanica is an annual food and oil crop native to Mexico and Guatemala. In this study, we report the complete chloroplast genome (plastome) of S. hispanica. The plastome was 150,980 bp in l...

B52 is a member of the classical serine/arginine (SR)-rich proteins, which are phylogenetically conserved and play significant roles in mRNA maturation, including alternative splicing. In the present study, the docking site, selector sequences and locus control region of the Chinese mitten crab (Eriocheir sinensis) Down syndrome cell adhesion molecule (EsDscam) were identified. Alternative splicing of Dscam is essential to generate different isoforms. We also isolated and characterised the B52 gene from E. sinensis (EsB52). The 876 bp open reading frame of EsB52 encodes a 291 amino acid residue polypeptide, and EsB52 has two RNA recognition motifs (RRMs) at the N-terminus and an arginine/serine-rich domain at the C-terminus. Each RRM contains two degenerate short submotifs, RNP-1 and RNP2. Analysis of tissue distribution revealed that EsB52 mRNA expression was widespread in all tested tissues, and especially high in brain and hemocytes. In hemocytes, EsB52 was upregulated significantly after stimulation with pathogen-associated molecular patterns and bacteria. Furthermore, EsB52 RNAi decreased the number of Ig7 inclusion in mRNA rather than Ig2 or Ig3. Taken together, these findings suggest that EsB52 acts as an alternative splicing activator of EsDscam.

Cleistogenes (Orininae, Cynodonteae, Chloridoideae, Poaceae) is an ecologically important genus. The phylogenetic placement of Cleistogenes and phylogenetic relationships among Cleistogenes taxa remain controversial for a long time. To resolve the intra- and inter-generic relationships of Cleistogenes, the plastomes of 12 Cleistogenes taxa (including 8 species and 4 varieties), one Orinus species, 15 Triodia species, two Tripogon species, and two Aeluropus species were included in the present study. All the taxa showed a similar pattern in plastome structure, gene order, gene content, and IR boundaries. The number of simple sequence repeats ranged from 145 (O. kokonorica) to 161 (T. plurinervata and T. schinzii). Moreover, 1,687 repeats were identified in these taxa, including 1,012 forward, 650 palindromic, 24 reverse, and one complement. Codon usage analysis revealed that these plastomes contained 16,633 (T. stipoides) to 16,678 (T. tomentosa) codons. Sequence divergence analysis among Cleistogenes and closely related genera identified five non-coding regions (trnS-UGA-psbZ, rpl32-trnL-UAG, trnQ-UUG-psbK, trnD-GUC-psbM, trnT-GGU-trnE-UUC). Phylogenetic analysis of complete plastomes indicated that Cleistogenes is sister to a clade composed of Orinus and Triodia, whereas it did not support the sister relationship between the recently proposed subtribe Orininae (Cleistogenes and Orinus) and Triodia. The subtribe Orininae was not supported by our complete plastome data. The split between Cleistogenes and Orinus-Triodia clade go back to 14.01 Ma. Besides, our findings suggested that C. squarrosa and C. songorica are the successive early diverging groups in the phylogenetic analysis. The other 10 taxa are divided into two groups: a monophyletic group composed of Cleistogenes sp. nov. and C. caespitosa var. ramosa is sister to other eight Cleistogenes taxa. Cleistogenes was estimated to have experienced rapid divergence within a short period, which could be a major obstacle in resolving phylogenetic relationships within Cleistogenes. Collectively, our results provided valuable insights into the phylogenetic study of grass species.

Tickborne intracellular bacterial pathogens including Anaplasma, Coxiella burnetti, Ehrlichia, and Rickettsia cause emerging infectious diseases worldwide. PCR was used to amplify the genes of these pathogens in Haemaphysalis flava ticks collected from hedgehogs in Central China. Among 125 samples including 20 egg batches, 24 engorged females, and 81 molted male and female adult ticks, the DNA sequences and phylogenetic analysis showed that the minimum infection rate of the ticks was 4% (5/125) for A. bovis, 3.2% (4/125) for C. burnetti, 9.6%, (12/125) for E. ewingii, and 5.6% for Rickettsia including R.japonica (3.2%, 4/125) and R. raoultii (2.4%, 3/125), respectively. The prevalence of these pathogens was significantly higher in dead engorged females (83.3%, 20/24) than in eggs (5%, 1/20) and molted ticks (8.6%, 7/81). Our study indicated that H. flava ticks could be infected with multiple species of tickborne pathogens including Anaplasma, C. burnetti, Ehrlichia, and Rickettsia in Central China, and the prevalence of these pathogens was reduced during transovarial and transstadial transmission in ticks, suggesting that ticks may not be real reservoirs but only vectors for these tickborne pathogens.

Eragrostideae Stapf, the second-largest tribe in Chloridoideae (Poaceae), is a taxonomically complex tribe. In this study, chloroplast genomes of 13 Eragrostideae species were newly sequenced and used to resolve the phylogenetic relationships within Eragrostideae. Including seven reported chloroplast genomes from Eragrostideae, the genome structure, number and type of genes, codon usage, and repeat sequences of 20 Eragrostideae species were analyzed. The length of these chloroplast genomes varied from 130,773 bp to 135,322 bp. These chloroplast genomes showed a typical quadripartite structure, including a large single-copy region (77,993&ndash, 80,643 bp), a small single-copy region (12,410&ndash, 12,668 bp), and a pair of inverted repeats region (19,394&ndash, 21,074 bp). There were, in total, 129&ndash, 133 genes annotated in the genome, including 83&ndash, 87 protein-coding genes, eight rRNA genes, and 38 tRNA genes. Forward and palindromic repeats were the most common repeat types. In total, 10 hypervariable regions (rpl22, rpoA, ndhF, matK, trnG&ndash, UCC-trnT&ndash, GGU, ndhF&ndash, rpl32, ycf4&ndash, cemA, rpl32&ndash, trnL&ndash, UAG, trnG&ndash, GCC&ndash, trnfM&ndash, CAU, and ccsA&ndash, ndhD) were found, which can be used as candidate molecular markers for Eragrostideae. Phylogenomic studies concluded that Enneapogon diverged first, and Eragrostis including Harpachne is the sister to Uniola. Furthermore, Harpachne harpachnoides is considered as a species of Eragrostis based on morphological and molecular evidence. In addition, the interspecies relationships within Eragrostis are resolved based on complete chloroplast genomes. This study provides useful chloroplast genomic information for further phylogenetic analysis of Eragrostideae.

Background Severe fever with thrombocytopenia syndrome virus (SFTSV) is a novel tick-borne phlebovirus, which is listed in the most dangerous pathogens by the World Health Organization, and has 12–30% fatality rates. SFTSV antibodies were reported in minks that experienced abortion or reproductive failure. The aim of this study was to determine whether SFTSV infection causes an adverse pregnancy outcome in the fetus using a pregnant mouse model. Methodology/Principal findings We found SFTSV in the fetus after infection in pregnant mice, and some dams showed adverse pregnancy outcomes after infection with SFTSV including placental damage, fetal reabsorption, and fetal intrauterine growth restriction (IUGR). SFTSV had obvious tropism characteristics in the placenta, especially in the labyrinth. In early-gestation, pregnant mice infected with SFTSV had fetal IUGR and a high viral load in the fetus. The virus widely spread in infected fetuses, including the hindbrain, thymus, heart, spinal cord, and liver. Conclusions Our study demonstrated that SFTSV was vertically transmitted to the fetus through the placental barrier of immunocompetent mice, and resulted in adverse pregnancy outcomes., Author summary Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease discovered in rural areas of China in 2009, caused by a novel tick-borne phlebovirus, the SFTS virus (SFTSV). At present, SFTS reported cases are increasing, and the epidemic area of SFTSV is expanding. SFTSV is currently endemic in China, Japan, South Korea and Vietnam, and has 12–30% fatality rates. This study demonstrated that SFTSV is vertically transmitted to the fetus through the placental barrier of immunocompetent mice, and can result in fetal damage. This study highlights the possible potential impact of SFTSV on pregnant women and livestock. The results suggested that we should actively monitor the pregnancy outcomes of women and livestock infected with SFTSV, and strengthen the study of the pathogenesis of fetal damage caused by SFTSV.

Currently, Zika virus (ZIKV) is spreading across the world and no ZIKV infection cases have ever been reported in China. Here, we aimed to determine whether ZIKV infection exists in China. Blood samples of 273 healthy individuals were collected from Nanning City, Guangxi Province, China in March 2019. We found that 9.5% (26/273) and 1.8% (5/273) of healthy persons were positive to ZIKV total antibody (IgG and/or IgM) IgM antibody, respectively. All ZIKV positive plasma samples were negative to Dengue virus and West Nile virus. Among the ZIKV antibody positive plasma samples, 65.4% (17/26) exhibited neutralizing activity to ZIKV. Followed up studies showed that none had clinical symptoms of ZIKV infection and oversea experience. Together, our study indicates that endemic ZIKV infections emerge in China, which not only suggested that ZIKV posed a potential threat to public health in China, but also expand the ZIKV epidemic areas in East and Southeast Asia., Author summary ZIKA, caused by mosquito-borne Zika virus (ZIKV), is typically asymptomatic or with mild fever, and the occurrence of ZIKV infections are reported around the world. However, limited ZIKV epidemiological investigation was performed and no ZIKV infection cases were ever reported in China. Here, we observed that Zika virus also distributes in China. This work not only suggests ZIKV as a potential threat to public health in China but also expands our knowledge of ZIKV epidemic regions in East and Southeast Asia. Therefore, government officials, physicians and public health workers in China need to pay attention to ZIKV infection cases.

As one of the most severe environmental stresses, salt stress can cause a series of changes in plants. In salt tolerant plant Zoysia macrostachya, germination, physiology, and genetic variation under salinity have been studied previously, and the morphology and distribution of salt glands have been clarified. However, no study has investigated the transcriptome of such species under salt stress. In the present study, we compared transcriptome of Z. macrostachya under normal conditions and salt stress (300 mmol/L NaCl, 24 h) aimed to identify transcriptome responses and molecular mechanisms under salt stress in Z. macrostachya. A total of 8703 differently expressed genes (DEGs) were identified, including 4903 up-regulated and 3800 down-regulated ones. Moreover, a series of molecular processes were identified by Gene Ontology (GO) analysis, and these processes were suggested to be closely related to salt tolerance in Z. macrostachya. The identified DEGs concentrated on regulating plant growth via plant hormone signal transduction, maintaining ion homeostasis via salt secretion and osmoregulatory substance accumulation and preventing oxidative damage via increasing the activity of ROS (reactive oxygen species) scavenging system. These changes may be the most important responses of Z. macrostachya under salt stress. Some key genes related to salt stress were identified meanwhile. Collectively, our findings provided valuable insights into the molecular mechanisms and genetic underpinnings of salt tolerance in Z. macrostachya.

Studies in E. sinensis have shown that ubiquitination mediated by Cullin-RING E3 ligases (CRLs) plays important roles in spermatogenesis. In other species, CRLs are also essential in cell cycle progression, DNA replication, signal transduction, gene transcription, and development. The catalytic RING component, the RING box protein, is an important part of CRLs. However, there have been few studies on CRLs in crustaceans. In this study, we cloned two RING box protein genes from the Chinese mitten crab, Eriocheir sinensis, termed Es-RBX1 and Es-RBX2 The full length Es-RBX1 cDNA comprises 741 nucleotides, and encodes a protein of 124 amino acid residues, whereas the Es-RBX2 cDNA comprises 1325 nucleotides, and encodes a protein of 110 amino acid residues. Bioinformatics analysis showed that the domains and structure of the RBX proteins have been highly conserved during evolution. Quantitative real-time polymerase chain reaction and western blotting showed that Es-RBX1 is highly expressed in the testis, particularly during the spermatocyte stage, whereas Es-RBX2 did not show specific expression in the male reproductive system. Furthermore, Es-RBX1 is mainly distributed in the nucleus, and changed its location with the development of the nucleus. Co-immunoprecipitation showed that Es-RBX1 could bind Cullin4. These results suggested that Es-RBX1 plays a key role in spermatogenesis of E. sinensis though forming a complex with Cullin4.

Salinity is one of the major environmental limitations to agricultural productivity throughout the world. To overcome the defects of microarray sequencing analyses caused by heterogeneity among ind...

PCR amplification of the rrs2 gene indicated that 50% (62/124) of insectivorous bats from eastern China were infected with Leptospira borgpetersenii, L. kirschneri, and several potentially new Leptospira species. Multilocus sequence typing defined 3 novel sequence types in L. kirschneri, suggesting that bats are major carriers of Leptospira.

Autophagy is essential for eukaryotic cell homeostasis and can perform both anti-viral and pro-viral roles depending on the kinds of viruses, cell types and cell environment. Severe fever with thrombocytopenia syndrome phlebovirus (SFTSV) is a newly discovered tick-borne virus in the Phenuiviridae family that causes a severe hemorrhagic fever disease in East Asia. In this study we determined interactions between SFTSV and autophagy. Our results showed that LC3-II (microtubule associated protein 1 light chain 3-II) protein accumulated from 4 h to 24 h after SFTSV infection compared to mock-infected Vero cells, and the use of E64d and pepstatin A did not affect the expression of LC3-II protein, which indicated that the increased LC3-II may be the result of inhibition of autophagic degradation caused by SFTSV infection. However, knockdown of LC3B promotes SFTSV replication, which indicated a negative role of LC3B protein in SFTSV replication. We also detected co-localization of SFTSV non-structure (NSs) protein with LC3B, p62 and Lamp2b respectively in SFTSV infected Vero cells, which indicated the possibility of selective autophagy or chaperone-mediated autophagy involving in SFTSV infection. Our results indicated that SFTSV infection promotes LC3 accumulation and several proteins of the autophagy pathway co-localize with NSs protein during SFTSV infection.

Background Pseudorabies, a highly contagious infectious disease of swine is caused by pseudorabies virus (PRV). PRV can cause fatal infection in other animal species. Results We report a deadly outbreak of pseudorabies that killed 87.2% (3522/4028) minks in a farm in 2014 in Shandong Province, China. PRV was isolated by using Vero cell culture and detected in mink samples by PCR from minks died during the outbreak. Epidemiological analysis indicated that 5.8% of minks (33/566) were PCR positive to PRV in Shandong Province. Phylogenetic analysis indicated that the PRV strains isolated from minks in this study were in the same clade with the Chinese porcine PRV isolates, which are resistant to the PRV vaccine. Conclusions We demonstrated that pseudorabies virus caused an outbreak of minks in a farm in Shandong Province of China and the virus has a very high infection rate in minks in Shandong Province, which is a challenge for the fur industry in China.

Scavenger receptors (SRs) are important pattern recognition receptors (PRRs), which play significant roles in host defense against pathogens by identifying pathogen-associated molecular patterns (PAMPs). In this study, we report the cloning and characterization of a SR from Eriocheir sinensis ( Es SR-B1) which is a 500 amino acid protein encoded by a gene comprised of 2726 nucleotides with a 1503 bp open reading frame. The domains of Es SR-B1 were found to be evolutionarily conserved. Es SR-B1 was widely detected in different tissues of E. sinensis and significantly up-regulated in hemocytes after stimulation by Staphyloccocus aureus or Vibrio parahaemolyticus . Recombinant Es SR-B1 protein could bind to bacteria and promote phagocytosis upon bacterial stimulation. Moreover, antimicrobial peptide expression was reduced in Es SR-B1-silenced hemocytes after challenge by S. aureus or V. parahaemolyticus . Thus, Es SR-B1 has a critical role in the binding of bacteria and subsequent promotion of hemocyte phagocytosis.

Vitellogenin (Vtg) is traditionally regarded as a key supplier of nutrients and energy during the early development of embryos and larvae, but accumulating evidence suggests that Vtg is also involved in innate immune defense. Whether Vtg is involved in innate immunity in Eriocheir sinensis, and its functions, remain largely unknown. In this study, a cDNA representing the vitellogenin1 gene from E. sinensis (Es-vtg1) was cloned. The full-length Es-vtg1 cDNA comprised 7939 nucleotides, encoding an open-reading frame of 2567 amino acid residues. Bioinformatic analysis showed that the domains of Es-Vtg1 have been conserved during evolution. Quantitative real-time PCR and western blotting showed that Es-vtg1 was highly expressed in ovary and hepatopancreas. Moreover, bacteria could induce the high-level expression of Es-Vtg1. Es-Vtg1 plays important roles in immunological defense, including binding to bacteria, inhibiting bacterial proliferation, and regulating the expression of antimicrobial peptides. Collectively, these results demonstrate that Es-Vtg1 plays critical roles in antimicrobial function.

We isolated Rickettsia japonica from a febrile patient in Lu’an City, China, in 2013. Subsequently, we found an R. japonica seroprevalence of 54.8% (494/902) in the rural population of Anhui Province and an R. japonica prevalence in Haemaphysalis longicornis ticks of 0.5% (5/935). R. japonica and its tick vector exist in China.

The complete chloroplast genome (plastome) of Maclura tricuspidate Carriere, a thorny and deciduous tree, is determined. The plastome is 161,348 bp in length, consisting of a 89,364 bp large single-copy region, a 20,246 bp small single-copy region, and two 25,869 bp inverted repeat regions. The GC content of this plastome is 36.1%. A total of 112 unique genes are annotated for the plastome of M. tricuspidata, containing 78 protein coding genes (PCGs), 30 tRNAs, and four rRNAs. Phylogenetic analysis shows that M. tricuspidata is sister to the clade comprising Dorstenieae, Castilleae, and Ficeae.

Eleusine coracana is a hardy crop that can grow in diverse environments. In this study, the complete plastome of E. coracana was determined. The plastome was 135,144 bp in size. It consists of a la...

The genomic and metabolic features of non-Saccharomyces yeast Nakazawaea ishiwadae GDMCC 60786 were investigated using its genome and by analyzing the metabolites of carbon source fermentation. N. ishiwadae GDMCC 60786, which showed resistant to environmental stresses (ethanol, SO2, pH, and NaCl) and a high ethyl acetate-production ability, was classified into the Nakazawaea genus based on the 26S rRNA gene. Complete genome sequences of the strain were obtained using the PacBio single-molecule real-time sequencing platform. The genome size was about 12,088,154 bp with 33.69% GC content. Key enzymes of glycolytic and pyruvate metabolism were identified by genes function annotation. KEGG analysis revealed that this strain possesses phosphoglucomutase, phosphofructokinase, phosphoglycerate kinase, phosphoglycerate mutase, pyruvate decarboxylase, and pyruvate dehydrogenase. Furthermore, combined with fermentation data on various carbon sources (e.g., glucose, fructose, maltose, and sucrose) and different amounts of ethanol and acetic acid, it was found that monosaccharides were more suitable as the carbon source of this strain to produce ethyl acetate. Compared with the control group, 4% (v/v) ethanol was used as supplementary carbon source, the content of ethyl acetate was increased 4 times.

Introduction: Recently, hantaviruses have been discovered in insectivores in Europe, Asia, Africa, and North America. Imjin virus (MJNV) was first isolated from the lung tissues of Ussuri white-toothed shrew (Crocidura lasiura) from South Korea in 2009. We aim to detect the species and prevalence of insectivore- and rodent-borne hantaviruses in shrews and rodents. Materials and Methods: Shrews and rodents were captured in Jiaonan County of Shandong Province, China, in 2014. RT-PCR was used to amplify viral RNA of Hantavirus species, including insectivore-borne Imjin virus (MJNV), rodent-borne Hantaan virus (HTNV), and Seoul virus (SEOV) from shrews and rodents. Results and Discussion: We found that MJNV infected 10.7% (19/178) of Crocidura shrews, but it infected none of rodents (0/475); we also found that 2 of 178 (1.1%) Crocidura shrews were PCR positive to SEOV. This study indicated that the major animal hosts of Imjin virus are shrews, and rodent-borne SEOV can infect shrews.

Salt stress adversely affects plant growth and productivity worldwide. Overcoming the defects of profiling analyses caused by heterogeneity of technical variations in individual studies and prediction of salt stress response key genes and pathways are conducive to mine potential salt responses mechanisms and of great interest for agricultural research. In this research, a literature-searching strategy was used to identify salt stress response expression profiling studies in Arabidopsis from the PubMed database, the Gene Expression Omnibus (GEO) database, and the ArrayExpress database. Twenty-five studies were filtered from the 2161, 121, and 700 identified studies. A rank aggregation method (RRA)-based meta-analysis were conducted to synthesize the 25 profiling datasets and identified 829 dysregulated genes with 452 up-regulated and 377 down-regulated in NaCl-treated samples. Gene ontology enrichment and pathway analyses classified the 452 up-regulated genes into functional categories involved “response to plant hormone and compounds,” “signaling” and “developmental plasticity,” etc. In total, 111 dysregulated transcription factors (TFs) were further identified and 387 differentially expressed genes (DEGs) were predicted as these TFs’ targets. The identification of transcriptional factors, functional repertoire, and biological pathways response to the salt stress signal would promote mechanistic studies underlying stress tolerance process in Arabidopsis at systematic level and would contribute to discover the conservation and diversification of stress resistance and adaptation abilities of various plants during the evolution of plant kingdom.

Leptotrombidium scutellare mites, the vector of Orientia tsutsugamushi, have rarely been reported to associate with Rickettsia species. Three hundred nineteen chiggers were collected from the ears of 32 rodents captured in Huangdao District of Qingdao City, China, in October 2015. The chigger samples were tested for Rickettsia, severe fever with thrombocytopenia syndrome virus, and hantavirus by PCR or RT-PCR amplification. All mites were classified morphologically and molecularly as L. scutellare chiggers. Rickettsial DNA sequences were amplified for four genes including 16S rRNA, ompB, gltA, and 17 kD protein genes. The minimum infection rate (MIR; number of positive pools/total specimens tested) of the Rickettsia species in the chiggers were 2.8% (9/319). Phylogenetic analysis indicated that individual genes were closely related to different Rickettsia species including R. felis (with 16S rRNA gene), R. australis (with gltA gene), an unnamed Rickettsia sp. TwKM02 (with ompB gene), and Rickettsia endosymbiont of soft tick Ornithodoros erraticus (with 17 kD protein gene). Phylogenic analysis of the concatenated sequence of 16S rRNA, gltA, ompB, and 17 kD protein genes indicated that the Rickettsia species from L. scutellare chigger was most closely related to R. australis and R. akari. These results indicated that the Rickettsia species in chiggers was unique; it was named Candidatus Rickettsia leptotrombidium. Severe fever with thrombocytopenia syndrome virus and hantavirus were not amplified from the chiggers, suggesting lack of infection of these pathogens in the chiggers. A unique Rickettsia species was detected in L. scutellare, which expanded the knowledge on the vector distribution of Rickettsia.

Studies in mammals have shown that damaged DNA-binding protein 1 (DDB1) is a multifunctional protein that recognizes UV-induced DNA lesions and activates nucleotide excision repair process, and could also be a linker protein for Cullin4 in ubiquitination to regulate cell cycle progression. However, there are few studies of DBB1 in crustaceans. In this study, a cDNA representing the DDB1 gene from Eriocheir sinensis (Es-DDB1) was cloned successfully. The full length Es-DDB1 cDNA comprises 4871 nucleotides, and encodes an open-reading frame (ORF) of 1137 amino acid residues. Bioinformatics' analysis showed that the domains and structure of Es-DDB1 have been highly conserved during evolution. Antibodies against Es-DDB1 and Es-Cul4 were raised using a prokaryotic expression system. Moreover, a co-immunoprecipitation assay showed that Es-DDB1 could bind Es-Cul4 in the testis of Eriocheir sinensis. Furthermore, quantitative real-time PCR and Western blotting showed high expression in the testis, particularly during the spermatocyte stage. Immunofluorescence assays showed that Es-DDB1 was mainly distributed in the cytoplasm in the early and middle developmental stages. These results indicated that Es-DDB1 might play a key role in spermatogenesis of E. sinensis.

Highlights • We found that 6.6% (13/197) bats from Shandong and Hubei provinces of China carried hepatis B virus (HBVs). • HBVs from bats in the two places were phylogenetically in the same cluster, but distinct from bat HBVs from other places. • HBVs were highly prevalent and genetic diversified in bats, supporting the hypothesis that bats may be the origin of primate hepadnaviruses., Bats have been identified as the hosts of hepatitis B virus (HBV) in recent years and bats HBV can infect human hepatocyte. We investigated the prevalence and genetic diversity of HBV in bats in China. In this study, a total of 197 insectivorous bats belonging to 10 bat species were captured from karst caves in Mengyin County, Shandong Province and Xianning City, Hubei Province, China. PCR amplification indicated that in total 6.6% (13/197) bats were positive to HBVs. The HBV positive rate in bats was 7.1% (9/127) and 5.7% (4/70) in Shandong Province and Hubei Province, respectively. Phylogenetic analysis indicated that HBV from the two places were in the same cluster with 90.5%–99.5% homology, but distinct from bat HBVs from other places in China and other countries. We concluded that HBV was prevalent and genetic diversified in bats, supporting the hypothesis that bats may be the origin of primate hepadnaviruses.

Pseudomonas aeruginosa, one of the most common pathogens in hospital-acquired infections, is tightly controlled by a multilayered regulatory network, including the quorum sensing system (QS), the type VI secretion system (T6SS), and resistance to host immunity. We found that the P. aeruginosa 3880 (PA3880) gene, which encodes an unknown protein, acts as a regulator of anaerobic metabolism in response to oxidative stress and virulence in P. aeruginosa. More than 30 PA3880 homologs were found in other bacterial genomes, indicating that PA3880 is widely distributed in the Bacteria kingdom as a highly conserved gene. Deletion of the PA3880 gene changed the expression levels of more than 700 genes, including a group of virulence genes, under both aerobic and anaerobic conditions. To further study the mechanisms of PA3880-mediated regulation in virulence, we utilized a bacterial two-hybrid assay and found that the PA3880 protein interacted directly with QS regulator MvfR and anaerobic regulator Anr. Loss of the PA3880 protein significantly blunted the pathogenicity of P. aeruginosa, resulting in increased host survival, decreased bacterial burdens, reduced inflammatory responses, and fewer lung injuries in challenged mice hosts. Mechanistically, we found that Cys44 was a critical site for the full function of PA3880 in influencing alveolar macrophage phagocytosis and bacterial clearance. We also found that AnvM directly interacted with host receptors Toll-like receptor 2 (TLR2) and TLR5, which might lead to activation of the host immune response. Hence, we gave the name AnvM (anaerobic and virulence modulator) to the PA3880 protein. This characterization of AnvM could help to uncover new targets and strategies to treat P. aeruginosa infections. IMPORTANCE Infections by Pseudomonas aeruginosa, one of the most frequently isolated human pathogens, can create huge financial burdens. However, knowledge of the molecular mechanisms involved in the pathogenesis of P. aeruginosa remains elusive. We identified AnvM as a novel regulator of virulence in P. aeruginosa. Deletion of anvM altered the expression levels of more than 700 genes under aerobic and anaerobic conditions, including quorum sensing system genes and oxidative stress resistance genes. AnvM directly interacted with MvfR and Anr, thus regulating their downstream genes. More importantly, AnvM directly bound to TLR2 and TLR5, which turn on the host immune response. These findings provide insights into the significance of AnvM homologs in pathogenic bacteria and suggest a potential drug target against bacterial infection.

PCR amplification indicated the minimum infection rate of Rickettsia spp. was 0.66% in Haemaphysalis longicornis ticks collected from Shandong Province, China. Phylogenetic analysis based on the rrs, gltA, ompA, and ompB genes indicated that the ticks carried R. japonica, Candidatus Rickettsia longicornii, and a novel Rickettsia species related to R. canadensis.

SummarySevere fever with thrombocytopaenia syndrome (SFTS) is an emerging infectious disease discovered in 2010 and has a case fatality as high as 30%. We intended to study the immune protection conferred by SFTS with natural infection. We collected and analysed 4-year follow-up data to study the characteristics of neutralising antibodies against SFTS virus (SFTSV). The 50% plaque reduction neutralisation test was used for the detection of neutralising antibodies against SFTSV. Geometric mean titres (GMTs) and proportions of patients with a protective titre were analysed, and the persistence of protection was predicted. The titre of antibodies declined yearly in the 4-year study period. Approximately 3 months after infection, the GMT was 143 (95% confidence interval (CI): 89–231), and 100% of patients had a protective titre. In the fourth year, the GMT declined to 53 (95% CI: 37–76), and 95% of patients had a protective titre. The titre was higher in females than in males. On average, the protection offered by neutralising antibodies against SFTSV could last as long as 9 years. The durations of protection were different for different initial titres. The characteristics of neutralising antibodies can be used as a reference for the vaccination doses and schedules of forthcoming vaccines.

Iron homeostasis is vital to organismal health; it is maintained by the iron regulatory protein (IRP)-iron-responsive element (IRE) signaling pathway. In the Chinese mitten crab Eriocheir sinensis, EsFer-1 and EsFer-2 reportedly have a putative IRE, but an IRP has not yet been identified. In this study, we successfully amplified the full-length cDNA of EsIRP using gene cloning and rapid amplification of cDNA ends techniques. The length of this cDNA was 4474 bp, and it included a 2682-bp open reading frame encoding 893 amino acids. Using quantitative real-time PCR, mRNA transcripts of EsIRP were detected in various tissues. The highest and lowest expression level was detected in the muscle and gills, respectively. In response to Staphylococcus aureus and Vibrio parahaemolyticus challenge, the transcription level of EsIRP was downregulated and that of EsFer-1 and EsFer-2 was upregulated in hemocytes. EsIRP knockdown resulted in increased expression of both EsFer-1 and EsFer-2. After EsFer-1 and EsFer-2 knockdown, the bacterial clearance ability of E. sinensis against S. aureus and V. parahaemolyticus was impaired. In conclusion, our results suggest that the IRP-IRE signaling pathway plays an important role in the innate immune system response in E. sinensis.

Background Severe fever with thrombocytopenia syndrome (SFTS) is an emerging viral hemorrhagic fever with a high fatality rate and high frequency of person-to-person transmission and is caused by SFTSV, a tick-borne Phlebovirus. Because SFTS has similar clinical manifestations and epidemic characters (such as spatial and temporal distributions) with hemorrhagic fever with renal syndrome (HFRS) in China, we reason that SFTS patients might be misdiagnosed as HFRS. Methodology/principal findings Acute-phase sera of 128 clinically diagnosed HFRS patients were retrospectively analyzed for Hantavirus IgM antibodies with ELISA. Hantavirus-negative patients’ sera were further analyzed for SFTSV IgM antibodies with ELISA. ELISA showed that 73 of 128 (57.0%) of clinically diagnosed HFRS patients were IgM antibody positive to Hantaviruses. Among the 55 Hantavirus-IgM negative patients, four (7.3%) were IgM antibody positive to SFTSV. The results indicated that the four SFTS patients were misdiagnosed as HFRS. The misdiagnosed SFTS patients had clinical manifestations common to HFRS and were unable to be differentiated from HFRS clinically. Conclusions Our study showed that SFTS patients could be clinically misdiagnosed as HFRS. The misdiagnosis of SFTS as HFRS causes particular concern because it may increase the risk of death of SFTS patients and person-to-person transmission of SFTSV without proper care for and isolation of SFTS patients.

Although bats were considered as a major host of trypanosomatid flagellates, information of trypanosomes in bats is unknown in China. We collected bats in 2015 from Shandong Province of China and used PCR to amplify the Trypanosoma glycosomal glyceraldehyde phosphate dehydrogenase (gGAPDH) gene and 18S rRNA gene from the bat blood samples and heart tissues. The results showed that 10.3% (13/126) of bats (Eptesicus serotinus and Myotis pequinius) were positive for trypanosomatid DNA and DNA sequencing showed that all PCR amplified Trypanosoma DNA belonged to T. dionisii. We concluded that T. dionisii had a infection rate in bats from China. For the first time, Trypanosoma infections were detected in bats from China, providing valuable information on the prevalence of these parasites in Asia. This is also the first report of Trypanosoma dionisii in Myotis pequinius, suggesting that Trypanosoma dionisii has a broad host species.

Salinity is one of the most important abiotic stresses threatening plant growth and agricultural productivity worldwide. In green alga Chlamydomonas reinhardtii, physiological evidence indicates that saline stress increases intracellular peroxide levels and inhibits photosynthetic-electron flow. However, understanding the genetic underpinnings of salt-responding traits in plantae remains a daunting challenge. In this study, the transcriptome analysis of short-term acclimation to salt stress (200 mM NaCl for 24 h) was performed in C. reinhardtii. A total of 10,635 unigenes were identified as being differently expressed by RNA-seq, including 5920 up- and 4715 down-regulated unigenes. A series of molecular cues were screened for salt stress response, including maintaining the lipid homeostasis by regulating phosphatidic acid, acetate being used as an alternative source of energy for solving impairment of photosynthesis, and enhancement of glycolysis metabolism to decrease the carbohydrate accumulation in cells. Our results may help understand the molecular and genetic underpinnings of salt stress responses in green alga C. reinhardtii.

Background Scrub typhus, caused by Orientia tsutsugamushi, an obligate intracellular gram-negative bacterium, along with hemorrhagic fever with renal syndrome (HFRS), caused by hantaviruses, are natural-focus infectious diseases prevalent in Shandong Province, China. Both diseases have similar clinical manifestations in certain disease stages and similar epidemic seasons, which has caused difficulties for physicians in distinguishing them. The aim of this study was to investigate whether misdiagnosis of scrub typhus as HFRS occurred in patients in Shandong Province. Methods Serum samples (N = 112) of clinically suspected HFRS patients from 2013 to 2014 in Shandong Province were analyzed with enzyme-linked immunosorbent assay (ELISA) for antibodies to both hantavirus and Orientia tsutsugamushi. Results ELISA showed that 56.3% (63/112) and 8.0% (9/112) of clinically suspected HFRS patients were IgM antibody positive to hantavirus and O. tsutsugamushi, respectively. Among the hantavirus IgM antibody positive patients, 7.9% (5/63) were also IgM antibody positive to O. tsutsugamushi. Among the hantavirus IgM antibody negative sera, 8.2% (4/49) of sera were positive to O. tsutsugamushi. Conclusions We concluded that some scrub typhus patients were misdiagnosed as HFRS and co-infection of scrub typhus and HFRS might exist in China. Due to the different treatments for scrub typhus and HFRS, physicians should carefully differentiate between scrub typhus and HFRS and consider administering anti-rickettsia antibiotics if treatment for HFRS alone does not work., Author summary Scrub typhus is caused by O. tsutsugamushi and hemorrhagic fever with renal syndrome (HFRS) is caused by hantavirus. Both diseases have similar manifestations and have their peak epidemic season in the fall, which makes their clinical differentiation difficult to be differentiated from each other clinically. In this study, we found that scrub typhus was indeed misdiagnosed as HFRS and co-infection of O. tsutsugamushi and hantaviruses might exist in China. Owing to different treatments of HFRS and scrub typhus, physicians need pay attention to differentiate them; identify co-infection of HFRS and scrub typhus; and consider administering antibiotics if antiviral treatment alone does not work.

As one of the most common abiotic stresses, salt stress seriously impairs crop yield. Brachypodium distachyon (L.) Beauv. is a model species for studying wheat and other grasses. In the present investigation, the physiological responses of B. distachyon treated with different concentrations of NaCl for 24 h were measured. Therefore, the control and the seedlings of B. distachyon treated with 200 mM NaCl for 24 h were selected for transcriptome analysis. Transcriptome differential analysis showed that a total of 4116 differentially expressed genes (DEGs) were recognized, including 3120 upregulated and 996 downregulated ones. GO enrichment assay indicated that some subsets of genes related to the active oxygen scavenging system, osmoregulatory substance metabolism, and abscisic-acid (ABA)-induced stomatal closure were significantly upregulated under salt stress. The MapMan analysis revealed that the upregulated genes were dramatically enriched in wax metabolic pathways. The expressions of transcription factor (TF) family members such as MYB, bHLH, and AP2/ERF were increased under salt stress, regulating the response of plants to salt stress. Collectively, these findings provided valuable insights into the mechanisms underlying the responses of grass crops to salt stress.

Melanization mediated by prophenoloxidase (proPO) activating system play an essential role in killing invading microorganisms in invertebrates. Lipopolysaccharide and β-1, 3-glucan binding protein (LGBP) as a pattern recognition protein have been demonstrated to active the proPO cascade in insect and shrimp. In this study, we investigated the role of LGBP in prophenoloxidase cascade-induced melanization in Chinese mitten crab (Eriocheir sinensis). By RT-PCR analysis, EsLGBP was detected in all tested tissues, and showed highest expression in hemocytes, gill, intestine and brain. The expression of EsLGBP was up-regulated in the hemocytes following injections of LPS and β-1, 3-glucan. The recombinant EsLGBP protein (rEsLGBP) was produced via prokaryotic expression system and affinity chromatography. By western blotting, rEsLGBP was discovered to exhibit the ability to bind to all tested microorganisms, including Gram-negative bacteria, Gram-positive bacteria and yeast (Pichia pastoris). Meanwhile we found rEsLGBP has a high binding activity towards microbial immune elicitors such as LPS and β-1, 3-glucan whereas no binding activity is detected with peptidoglycan. Moreover, the effects of RNAi-mediated blockade of EsLGBP were investigated on bacterial counts in the hemolymph and cumulative mortality rate of crabs infected with Vibrio parahaemolyticus in vivo. Further experiments demonstrate that rEsLGBP can trigger the whole hemolymph dependent melanization and stimulate to proPO cascade in vitro. Taken together, these results provide experimental evidence for role of LGBP in innate immunity, especially in the activation of prophenoloxidase activating system.

Glycogen synthase kinase3β (GSK3β) is a Serine/Threonine kinase involved in metabolic reactions, and the Wnt, Akt, Hedgehog and BMP/Smad1 signalling pathways. It regulates cell metabolism, the cell cycle and cell fate through the phosphorylation of a diverse array of substrates. Its disorders lead to testicular degeneration, testicular cord disruption and many other reproductive diseases that often lead to infertility. In mammals, GSK3β plays an important role in the regulation of spermatogenesis. To explore functions of GSK3β during spermatogenesis inEriocheir sinensiswe cloned the full-lengthGSK3β cDNA sequence from the testis ofE. sinensis(EsGSK3β). The 1161-nucleotide open reading frame encodes a 386-amino-acid protein with a predicted molecular mass of 43 kDa. Additionally, we examined the expression pattern of EsGSK3β in different tissues and testis developmental stages using real-time PCR and Western blotting. Localization analysis ofEsGSK3β RNA and protein by in situ hybridization and immunofluorescence, respectively, showed that molecules translocated from the cytoplasm and nucleus to the regions of the acrosomal tubule and apical cap during spermatogenesis. The results indicated that EsGSK3β might play an important role duringE. sinensisspermatogenesis.

Transstadial and transovarial virus transmission occur among ticks, and transmission to mice can occur through a tick bite., Severe fever with thrombocytopenia syndrome (SFTS) is an emerging hemorrhagic fever in East Asia caused by SFTS virus (SFTSV), a newly discovered phlebovirus. The Haemaphysalis longicornis tick has been suspected to be the vector of SFTSV. To determine whether SFTSV can be transmitted among ticks, from ticks to animals, and from animals to ticks, we conducted transmission studies between developmental stages of H. longicornis ticks and between ticks and mice. Using reverse transcription PCR, we also analyzed the prevalence of SFTSV infection among H. longicornis ticks collected from vegetation in Shandong Province, China. Our results showed a low prevalence of SFTSV among collected ticks (0.2%, 8/3,300 ticks), and we showed that ticks fed on SFTSV-infected mice could acquire the virus and transstadially and transovarially transmit it to other developmental stages of ticks. Furthermore, SFTSV-infected ticks could transmit the virus to mice during feeding. Our findings indicate ticks could serve as a vector and reservoir of SFTSV.

Dodders (Cuscuta spp.) are widespread parasitic weeds on many plants all over the world, which can lead to considerable damage to ecosystems (Costea et al. 2011). Many Cuscuta species are recognized as class 1 designated Prohibited Noxious Weeds (Government of Canada 2016), because they can reduce the biomass of hosts and modify the structure of plant communities (Pennings and Callaway 2002). Cuscuta japonica is a hemiparasitic plant with many hosts including Araliaceae, Asteraceae, Fabaceae, Polygonaceae, Rutaceae, and Salicaceae (Chen et al. 2012; Yuan et al. 2015). Until to now, C. japonica has not been reported parasitizing hosts from gymnosperm species. In September 2019, dodder was observed parasitizing Pinus densiflora in Lushan Mountain (36°18′17.99″N, 118°5′50.27″E, elevation 662 m), Zibo City, Shandong Province, China. P. densiflora, commonly known as Japanese red pine, is an evergreen pine with a range including Japan, the Korean Peninsula, and northeastern China. It is widely cultivated due to its timber for construction, trunk for resin, bark for tannin, and needles for medicine. One individual of P. densiflora, approximately 5 m tall, was infected, and the branches and needles of the host were wrapped by the leafless stems of this dodder. The haustoria penetrating branches and needles of the host were visible to the naked eye. Uneven holes emerged on Pinus needle after removal of the dodder stem, and the tissues of Pinus needle around the holes became black. Some needles were chlorotic, wilted, and even defoliated. This dodder can be morphologically identified as C. japonica according to previous investigation records and descriptions, including slightly yellow stems with branches 1.4 to 2.6 mm in diameter, capsules globose and smooth, one style, and stigma two-lobed with elongated lobes. An area of ∼5.8 ha around the infected P. densiflora was carefully examined for this dodder, and only three infections were discovered on one individual of P. densiflora. No dodders were found on the nearby deciduous trees within the above examined area. However, this dodder was found parasitizing deciduous trees such as Ulmus pumila outside of the examined area in the Lushan mountain, so P. densiflora might not be a primary host for this dodder from the point of view of the entire mountain. Therefore, we tentatively classify P. densiflora as a rare host of this dodder. Species identification was further conducted by characterizing the whole plastome. First, total DNA was extracted from silica-dried stems of this dodder. Then the genome-skimming method was applied to generate the whole plastome as described in Qu et al. (2019). One sequence of 121,005 bp was deposited in GenBank (accession no. MN585289). BLAST analysis reconfirmed the identity of this dodder as C. japonica (99.92%, target accession no. MH780080). Furthermore, phylogenetic analysis supported this dodder as sister to C. japonica with nearly the same branch length (0.0002 versus 0.0003) with 100% bootstrap support value. Thus, this dodder observed in Zibo City was identified as C. japonica based on evidence from morphology, sequence, and phylogeny. Specimens of C. japonica on P. densiflora were collected and deposited at the herbarium of the College of Life Sciences, Shandong Normal University (no. 092217A). C. japonica seems to be an extremely rare occurrence on P. densiflora. It will need more evidence to judge if it is a real infection on P. densiflora. It is probable that C. japonica is not a parasite that can threaten populations of P. densiflora at present. Therefore, C. japonica is not considered as a management concern on Japanese red pine. To the best of our knowledge, this is the first report of C. japonica parasitizing P. densiflora, and the only known example of C. japonica parasitizing a gymnosperm species in the world.

Background Timely diagnosis of SARS-CoV-2 infection is a prerequisite for treatment and prevention. The serology characteristics and complement diagnosis value of the antibody test to RNA test need to be demonstrated. Method Serial sera of 80 patients with PCR-confirmed COVID-19 were collected at the First Affiliated Hospital of Zhejiang University, China. Total antibody (Ab), IgM and IgG antibodies against SARS-CoV-2 were detected, and the antibody dynamics during the infection were described. Results The seroconversion rates for Ab, IgM and IgG were 98.8%, 93.8% and 93.8%, respectively. The first detectible serology marker was Ab, followed by IgM and IgG, with a median seroconversion time of 15, 18 and 20 days post exposure (d.p.e) or 9, 10 and 12 days post onset (d.p.o), respectively. The antibody levels increased rapidly beginning at 6 d.p.o. and were accompanied by a decline in viral load. For patients in the early stage of illness (0–7 d.p.o), Ab showed the highest sensitivity (64.1%) compared to IgM and IgG (33.3% for both, p, Antibody responses were induced after SARS-CoV-2 infection, and the complementary diagnostic value of antibody test to RNA test was observed. Antibody tests are critical tools in clinical management and control of SARS-CoV-2 infection and COVID-19.