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1.
Fish Shellfish Immunol ; 97: 432-439, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31883470

RESUMO

As a high mortality disease, Infectious spleen and kidney necrosis virus (ISKNV) can cause massive economic damage on mandarin fish farming industry in China, which seriously hindered the development of mandarin fish farming industry. In this research, SWCNTs (single-walled carbon nanotubes) as a candidate for DNA vaccine carrier was vaccinated by immersion (1, 2, 5, 10, 20 mg/L) in juvenile mandarin fish. In muscle, spleen and kidney tissues, the results showed that transcription and expression of MCP gene can be detected in pcDNA-MCP and SWCNTs-pcDNA-MCP groups after bath immunization. The immune response (immune-related genes expression, serum antibody production, enzyme activities and C3 content) was significantly enhanced in fish which vaccinated with SWCNTs-pcDNA-MCP in comparison with those vaccinated with pcDNA-MCP alone. After 14 d challenge, the RPS (relative percentage survival) can be enhanced which using SWCNTs as a carrier in SWCNTs-pcDNA-MCP (82.4%) group at 20 mg/L (the highest vaccine dose) than the naked pcDNA-MCP (54.2%) group. This study reveals that functionalized SWCNTs could be a promising immersion DNA vaccine carrier in aquaculture.

2.
Fish Shellfish Immunol ; 92: 133-140, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31173860

RESUMO

Infectious spleen and kidney necrosis virus (ISKNV) cause a high mortality disease which lead to significant economic loss on mandarin fish in China. There is no effective drug or vaccine against this fatal disease at present. Meanwhile, many drugs and vaccines had no effect in many cases account of several impenetrable barriers (cell, skin and gastrointestinal tract). Here we reported an immersion subunit vaccine system (SWCNTs-MCP) encoding MCP gene of ISKNV based on single-walled carbon nanotubes (SWCNTs). To evaluate its efficacy against ISKNV, we found a stronger and longer duration immune response (serum antibody production, enzyme activities and immune-related genes expression) can be induced in fish vaccinated with SWCNTs-MCP in comparison with those vaccinated with MCP alone. Importantly, SWCNTs can increase the immune protective effect of naked subunit vaccine by ca. 23.8%. Thereby, this study demonstrates that SWCNTs as a promising carrier for subunit vaccine might be used to vaccinate large-scale juvenile mandarin fish by bath administration approach.


Assuntos
Doenças dos Peixes/imunologia , Imunidade Inata , Iridoviridae/imunologia , Nanotubos de Carbono , Perciformes/imunologia , Vacinação/veterinária , Vacinas Virais/imunologia , Animais , Infecções por Vírus de DNA/imunologia , Infecções por Vírus de DNA/veterinária , Vacinas de Subunidades/imunologia
3.
BMC Microbiol ; 18(1): 113, 2018 09 10.
Artigo em Inglês | MEDLINE | ID: mdl-30200876

RESUMO

BACKGROUND: Fish culture in rice paddies can contribute to increasing yields of rice and surplus fish products. Environmental impacts and food-safety issues have become important topics in aquaculture, and organic foods currently were paid attention by researchers and industry practitioners. But the mechanism of differences in quality of Loach (Paramisgurnus dabryanus) reared in rice fields and ponds remains largely uncharacterized. In this study,digestive enzyme activity, intestinal mucosa cells and the gut microbial community of loach were determined under the two separate cultivation modes. RESULTS: The levels of intestinal digestive enzyme activity of fish reared in the paddy-cultivated mode (PACM) were higher (P < 0.05) than those in the pond-cultivated mode (POCM). It was extremely significant (P < 0.01) for the activity of lipase in the liver, foregut and midgut, and for the activities of amylase and trypsin in the hindgut. Acid mucous cells in the loach foregut in PACM were fewer than in POCM (P < 0.01). In summer, the abundance of the Firmicutes, Lactobacillus spp., Aeromonas hydrophila, Enterobacteriaceae and Streptococcus spp. in loach intestinal mucosa in PACM was higher than in POCM. In fall, the abundance of total bacteria, the Bacteroidetes, Bifidobacterium and Enterobacteriaceae in the intestinal mucosa in PACM was likewise higher than in POCM. These differences were significant (P < 0.05 or P < 0.01) between loach in the two separate culture modes for all microorganisms except for A. hydrophila and Streptococcus spp. In addition, quantitative PCR assays showed that some microorganisms presented consistently similar abundances in the gut as in the culture water. CONCLUSIONS: These results showed some enzymatic activities involved in digestion in liver and intestine of loach in PACM were higher than those in POCM, as using digestive enzyme analysis and histological observation of intestinal sections. These findings suggest most of the microorganisms examined in the gut mucosa of loach in the two culture modes significantly differed in abundance between summer and fall. However, some pathogenic bacteria in the gut, particularly A. hydrophila, presented lower abundance in PACM in fall, yet did not differ in abundance between loach in the two cultivation modes.


Assuntos
Amilases/metabolismo , Bactérias/isolamento & purificação , Cipriniformes/microbiologia , Microbioma Gastrointestinal , Mucosa Intestinal/enzimologia , Tripsina/metabolismo , Animais , Aquicultura , Bactérias/classificação , Bactérias/genética , Cipriniformes/crescimento & desenvolvimento , Mucosa Intestinal/microbiologia , Estações do Ano
4.
Fish Shellfish Immunol ; 64: 104-110, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28263880

RESUMO

Tissue factor pathway inhibitor 2 (TFPI-2) is an analog of TFPI-1 and a potent endogenous inhibitor of tissue factor (TF)-mediated blood coagulation. Recent reports have proven that the C-terminal of TFPI-2 peptides in humans and several other vertebrates possesses antibacterial activity against Gram-positive and Gram-negative bacteria. In our previous study, we reported that the TFPI-2 peptide, TC38 in tongue sole (Cynoglossus semilaevis) was active against Micrococcus luteus. In this study, we further examine the antimicrobial spectrum, mechanism of action, and function of TC38 in tongue sole. Our results indicate that TC38 is active against the Gram-negative bacteria Vibrio ichthyoenteri, Vibrio litoralis, Vibrio parahaemolyticus, and Vibrio vulnificus, as well as the fish Megalocytivirus, infectious spleen and kidney necrosis virus (ISKNV). The mechanism of action of TC38 against V. vulnificus was explored. The results showed that TC38 killed V. vulnificus cells without lysis of the cell membrane. FITC-labeled TC38 was able to penetrate the cell membrane and bind to DNA and RNA, then disrupt cellular function, eventually leading to cell death. Administration of TC38 to tongue sole significantly improved its defense against V. vulnificus infection. Overall, these results indicate that TC38 is a novel peptide with a broad antimicrobial spectrum. Furthermore, the unique action of TC38 against V. vulnificus adds new insights to the mechanism of action of vertebrate TFPI peptides. Moreover, TC38 is an interesting antimicrobial agent that could be useful in the fight against pathogenic invasion in aquaculture.


Assuntos
Infecções por Vírus de DNA/veterinária , Doenças dos Peixes/genética , Proteínas de Peixes/genética , Linguados , Glicoproteínas/genética , Vibrioses/veterinária , Animais , Membrana Celular/microbiologia , Membrana Celular/virologia , Infecções por Vírus de DNA/genética , Infecções por Vírus de DNA/imunologia , Infecções por Vírus de DNA/virologia , Doenças dos Peixes/imunologia , Doenças dos Peixes/microbiologia , Doenças dos Peixes/virologia , Proteínas de Peixes/metabolismo , Glicoproteínas/metabolismo , Iridoviridae/fisiologia , Ácidos Nucleicos/metabolismo , Distribuição Aleatória , Vibrio/fisiologia , Vibrioses/genética , Vibrioses/imunologia , Vibrioses/microbiologia
5.
Fish Shellfish Immunol ; 60: 466-473, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-27840169

RESUMO

Tissue factor pathway inhibitor 1 (TFPI-1) is a serine protease inhibitor that inhibits tissue factor (TF)-mediated coagulation. The C-terminal region of TFPI-1 could be cleaved off and proved to be antimicrobial against a broad-spectrum of microorganism. In a previous study, a C-terminal peptide, TC24 (with 24 amino acids), derived from tongue sole (Cynoglossus semilaevis) TFPI-1, was synthesized and found antibacterial against Micrococcus luteus. In the present study, the antibacterial spectrum and the action mode of TC24 was further examined, and its in vivo function was analyzed. Our results showed that TC24 also possesses bactericidal activity against Staphylococcus aureus and Vibrio vulnificus. During its interaction with the target bacterial cells, TC24 destroyed cell membrane integrity, penetrated into the cytoplasm, and induced degradation of genomic DNA and total RNA. In vivo study showed that administration of tongue sole with TC24 before bacterial and viral infection significantly reduced pathogen dissemination and replication in tissues. These results indicated that TC24 is a novel antimicrobial peptide against bacterial and viral pathogens, and that the observed effect of TC24 on bacterial RNA adds new insights to the action mechanism of fish antimicrobial peptides. Moreover, TC24 may play an important role in fighting pathogenic infection in aquaculture.


Assuntos
Doenças dos Peixes/genética , Proteínas de Peixes/genética , Linguados , Lipoproteínas/genética , Animais , Infecções por Vírus de DNA/genética , Infecções por Vírus de DNA/imunologia , Infecções por Vírus de DNA/veterinária , Infecções por Vírus de DNA/virologia , Doenças dos Peixes/imunologia , Doenças dos Peixes/microbiologia , Doenças dos Peixes/virologia , Proteínas de Peixes/metabolismo , Bactérias Gram-Negativas/fisiologia , Infecções por Bactérias Gram-Negativas/genética , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/veterinária , Bactérias Gram-Positivas/fisiologia , Infecções por Bactérias Gram-Positivas/genética , Infecções por Bactérias Gram-Positivas/imunologia , Infecções por Bactérias Gram-Positivas/microbiologia , Infecções por Bactérias Gram-Positivas/veterinária , Iridoviridae/fisiologia , Lipoproteínas/metabolismo , Análise de Sequência de DNA/veterinária
6.
Fish Shellfish Immunol ; 57: 309-317, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27569982

RESUMO

C-type lectins (CTLs) are important pattern recognition receptors (PRRs) that play vital roles in innate immunity. In teleosts, a number of CTLs have been reported, but their in vivo effects on host defense are still limited. In this study, a CTL homolog (SsLec1) was identified from black rockfish, Sebastes schlegelii, and its structure, expression and biological function was analyzed. The open reading frame of SsLec1 is 633 bp, with a 5'- untranslated region (UTR) of 36 bp and a 3'- UTR of 117 bp. The deduced amino acid sequence of SsLec1 shares the highest overall identity (73.20%) with the CTL of Oplegnathus fasciatus. SsLec1 possesses conserved CTL features, including a carbohydrate-recognition domain, four disulfide bond-forming cysteine residues, the mannose-type carbohydrate-binding motif, the conserved calcium binding sites and a putative signal peptide. The expression of SsLec1 was highest in liver and could be induced by experimental infection with Listonella anguillarum. Recombinant SsLec1 (rSsLec1) purified from E. coli was able to bind and agglutinate the Gram-negative fish pathogens Vibrio ichthyoenteri and Vibrio vulnificus. The agglutinating ability of rSsLec1 was abolished in the presence of mannose or ethylenediaminetetraacetic acid. Further analysis showed that rSsLec1 could enhance phagocytosis by macrophages. In vivo experiments indicated that rSsLec1 could inhibit bacterial infection and promote viral invasion. Taken together, these results suggest that SsLec1 is a novel CTL that possesses apparent immunoregulation property and plays a critical role in host defense against pathogens invasion.


Assuntos
Doenças dos Peixes/genética , Proteínas de Peixes/genética , Peixes , Imunidade Inata , Lectinas Tipo C/genética , Vibrioses/veterinária , Vibrio/fisiologia , Sequência de Aminoácidos , Animais , Clonagem Molecular , DNA Complementar/genética , DNA Complementar/metabolismo , Doenças dos Peixes/imunologia , Doenças dos Peixes/microbiologia , Proteínas de Peixes/química , Proteínas de Peixes/metabolismo , Lectinas Tipo C/química , Lectinas Tipo C/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Alinhamento de Sequência/veterinária , Vibrioses/genética , Vibrioses/imunologia , Vibrioses/microbiologia
7.
Yi Chuan ; 37(7): 702-10, 2015 07.
Artigo em Chinês | MEDLINE | ID: mdl-26351170

RESUMO

As a key component of life science, bioinformatics has been widely applied in genomics, transcriptomics, and proteomics. However, the requirement of high-performance computers rather than common personal computers for constructing a bioinformatics platform significantly limited the application of bioinformatics in aquatic science. In this study, we constructed a bioinformatic analysis platform for aquatic pathogen based on the MilkyWay-2 supercomputer. The platform consisted of three functional modules, including genomic and transcriptomic sequencing data analysis, protein structure prediction, and molecular dynamics simulations. To validate the practicability of the platform, we performed bioinformatic analysis on aquatic pathogenic organisms. For example, genes of Flavobacterium johnsoniae M168 were identified and annotated via Blast searches, GO and InterPro annotations. Protein structural models for five small segments of grass carp reovirus HZ-08 were constructed by homology modeling. Molecular dynamics simulations were performed on out membrane protein A of Aeromonas hydrophila, and the changes of system temperature, total energy, root mean square deviation and conformation of the loops during equilibration were also observed. These results showed that the bioinformatic analysis platform for aquatic pathogen has been successfully built on the MilkyWay-2 supercomputer. This study will provide insights into the construction of bioinformatic analysis platform for other subjects.


Assuntos
Biologia Computacional/métodos , Computadores , Aeromonas hydrophila/química , Animais , Proteínas da Membrana Bacteriana Externa/química , Carpas/virologia , Flavobacterium/genética , Simulação de Dinâmica Molecular , Reoviridae/genética , Proteínas Virais/química
8.
Bing Du Xue Bao ; 23(5): 412-6, 2007 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-17969861

RESUMO

The major capsid protein of lymphocystis disease virus isolated from Rachycentron canadum (LCDV-rc) was amplified and analysed. The 457bp DNA core fragment was amplified with the degenerate primers designed according to the conserved sequences of MCP gene of iridoviruses, then the flaking sequences adjacent to the core region were amplified by inverse PCR, and the complete sequence was obtained by combining all of them. The open reading frame of the gene is 1380bp in length, encoding a putative protein of 459 aa with molecular weight 51.12 kD and pI 6.87. Constructing the phylogenetic tree for comparing the MCP amino acid of iridoviruses, the results indicated that LCDV-rc is most homologous to the other Lymphocystis viruses and all of them constitute a branch. Accordingly LCDV-rc is identified as Lymphocystivirus.


Assuntos
Proteínas do Capsídeo/genética , Iridoviridae/genética , Perciformes/microbiologia , Reação em Cadeia da Polimerase/métodos , Animais , Sequência de Bases , Iridoviridae/classificação , Dados de Sequência Molecular , Filogenia
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