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1.
Fish Shellfish Immunol ; 93: 531-541, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31369858

RESUMO

Aquaculture is one of the world's most important and fastest growing food production sectors, with an average annual growth of 5.8% during the period 2001-2016. Common carp (Cyprinus carpio) is one of the main aquatic species produced for human consumption and is the world's third most produced finfish. Koi carp, on the other hand, are grown as a popular ornamental fish. In the late 1990s, both of these sectors were threatened by the emergence of a deadly disease caused by cyprinid herpesvirus 3 (CyHV-3; initially called koi herpesvirus or KHV). Since then, several research groups have focused their work on developing methods to fight this disease. Despite increasing knowledge about the pathobiology of this virus, there are currently no efficient and cost-effective therapeutic methods available to fight this disease. Facing the lack of efficient treatments, safe and efficacious prophylactic methods such as the use of vaccines represent the most promising approach to the control of this virus. The common carp production sector is not a heavily industrialized production sector and the fish produced have low individual value. Therefore, development of vaccine methods adapted to mass vaccination are more suitable. Multiple vaccine candidates against CyHV-3 have been developed and studied, including DNA, bacterial vector, inactivated, conventional attenuated and recombinant attenuated vaccines. However, there is currently only one vaccine commercially available in limited regions. The present review aims to summarize and evaluate the knowledge acquired from the study of these vaccines against CyHV-3 and provide discussion on future prospects.


Assuntos
Carpas/imunologia , Doenças dos Peixes/prevenção & controle , Herpesviridae/imunologia , Vacinas contra Herpesvirus/imunologia , Animais , Doenças dos Peixes/imunologia , Infecções por Herpesviridae/imunologia , Infecções por Herpesviridae/prevenção & controle , Infecções por Herpesviridae/veterinária
2.
Fish Shellfish Immunol ; 93: 669-682, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31408728

RESUMO

Iron is an important mineral element for fish. In this study, we investigated the influences of dietary iron deficiency on intestinal immune function as well as underlying signaling of on-growing grass carp (Ctenopharyngodon idella). Fish were fed with six graded level of dietary iron for sixty days, and a fourteen days' challenge test under infection of Aeromonas hydrophila thereafter. Results showed that compared with optimal iron level, iron deficiency increased enteritis morbidity, decreased lysozyme (LZ) and acid phosphatase (ACP) activities, complement 3 (C3), C4 and immunoglobulin M (IgM) concentrations and down-regulated mRNA levels of hepcidin, liver expressed antimicrobial peptide 2A (LEAP-2A), LEAP-2B, Mucin2, ß-defensin-1, anti-inflammatory cytokines transforming growth factor ß1 (TGF-ß1), TGF-ß2, interleukin 4/13A (IL-4/13A), IL-4/13B, IL-10, IL-11 and IL-15, inhibitor of κBα (IκBα), target of rapamycin (TOR) and ribosomal protein S6 kinase 1 (S6K1), whereas up-regulated mRNA levels of pro-inflammatory cytokines IL-1ß, interferon γ2 (IFN-γ2), IL-8, IL-12p35, IL-12p40 and IL-17D, nuclear factor kappa B (NF-κB) p65, IκB kinases α (IKKα), IKKß and eIF4E-binding protein (4E-BP) in intestine of on-growing grass carp, indicating that iron deficiency impaired intestinal immune function of fish under infection of A. hydrophila. Besides, iron excess also increased enteritis morbidity and impaired immune function of fish under infection of A. hydrophila. In addition, the effect of ferrous fumarate on intestinal immune function of on-growing grass carp is more efficient than ferrous sulfate. Finally, based on ability against enteritis, LZ activities in mid intestine and distal intestine, we recommended adding 83.37, 86.71 and 85.39 mg iron/kg into diet, respectively.


Assuntos
Carpas/imunologia , Doenças dos Peixes/imunologia , Proteínas de Peixes/metabolismo , Intestinos/imunologia , Ferro/deficiência , Transdução de Sinais/efeitos dos fármacos , Aeromonas hydrophila/fisiologia , Animais , Peptídeos Catiônicos Antimicrobianos/metabolismo , Citocinas/metabolismo , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/veterinária , Intestinos/efeitos dos fármacos , Ferro na Dieta/metabolismo , NF-kappa B/metabolismo , Distribuição Aleatória , Serina-Treonina Quinases TOR/metabolismo
3.
J Appl Microbiol ; 127(5): 1327-1338, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31373737

RESUMO

AIMS: An 8-week trial was performed to evaluate the differences in the intestinal microbiota and immune function of common carp (Cyprinus carpio)-fed different diets. METHODS AND RESULTS: The fish (initial weight 492·6 ± 30 g, n = 270) were randomly divided into three groups with three replicates and were fed earthworms (group A), earthworms + duckweed (group M) or duckweed (group P) respectively. The diversity and dominant microbiota of the intestinal bacteria were detected by denaturing gradient gel electrophoresis, and the abundance of dominant bacteria was quantified by qPCR. Additionally, the activities of some nonspecific immune enzymes and antioxidant enzymes were determined. The results showed that higher diversity and abundance of intestinal microbiota were observed in group M and group P (P < 0·05). Based on the intestinal microbiota, Cetobacterium was only detected in the intestines of common carp in group A, and Bacillus was identified in groups M and P. Additionally, a higher abundance of Bacteroidetes and Firmicutes was also found in the intestine in group P than in group A (P < 0·05). Interestingly, the higher activities of immune enzymes were detected in intestine of common carp in group M, such as acid phosphatase, phosphatase (AKP), lysozyme, total antioxidant capacity, superoxide dismutase, catalase, glutathione peroxidase. In addition, the lower level of metabolites were also detected, such as nitric oxide and malondialdehyde. CONCLUSIONS: Our results indicate that the intestinal microbiota and intestinal immunity of common carp were affected by diet. Meanwhile, the results show that a mixed diet can promote and improve the immune function of the omnivorous carp intestine, which suggests that paddy fields might be more suitable for the intestinal health and animal welfare of omnivorous fishes because they contain plant and animal diets. SIGNIFICANCE AND IMPACT OF THE STUDY: As an ecological aquaculture strategy, the rice-fish mode has attracted attention among farmers, researchers and even consumers, especially for the cultivation of common carp in paddy fields. In paddy fields, fish can eat plant- and animal-based diets. However, it is not clear whether common carp feeding on a mixed diet in paddy fields have better intestinal health. This experiment is one of only a few studies performed from the perspective of intestinal micro-organisms and immunity to successfully study the effects of different natural diets on adult common carp. This study also provides a theoretical basis for healthy breeding of common carp in paddy fields.


Assuntos
Bactérias/imunologia , Carpas/imunologia , Carpas/microbiologia , Dieta/veterinária , Microbioma Gastrointestinal/imunologia , Intestinos/imunologia , Intestinos/microbiologia , Ração Animal/análise , Animais , Antioxidantes/metabolismo , Aquicultura , Bactérias/classificação , Bactérias/genética , Bactérias/metabolismo , Microbioma Gastrointestinal/genética
4.
Microb Pathog ; 135: 103649, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31374321

RESUMO

Extracellular adenosine triphosphate (ATP) is as key mediator of immune and inflammatory responses. ATP is normally sequestered in the intracellular milieu and released by apoptotic and necrotic cells, where it acts as a pro-inflammatory mediator in the extracellular milieu. A limited number of studies have explored the involvement of purinergic signaling in oomycete infections, including Saprolegnia parasitica; this is a most destructive oomycete pathogen, associated with high mortality and severe economic losses for fish producers. The aim of this study was to determine whether purinergic signaling exerts anti- or pro-inflammatory effects in spleens of grass carp (Ctenopharyngodon idella) naturally infected by S. parasitica. Animals naturally infected with S. parasitica showed typical gross lesions characterized by cotton-wool tufts on the tail and fins, as well as severe histopathological lesions such as necrosis. Spleen ATP and metabolites of nitric oxide (NOx) levels were higher in fish naturally infected by S. parasitica compared to control on day 7 post-infection (PI). Spleen nucleoside triphosphate diphosphohydrolase (NTPDase) activity (ATP as substrate) was greater in fish naturally infected by S. parasitica than in uninfected on day 7 PI, while no significant differences were observed between groups with respect to NTPDase (adenosine diphosphate as substrate) and 5'-nucleotidase activities. Finally, adenosine deaminase (ADA) activity was lower in fish naturally infected by S. parasitica than in uninfected fish on day 7 PI. In summary, spleen tissue necrosis in the context of saprolegniosis provokes an intense release of ATP into the extracellular milieu, where it interacts with the P2X7 purine receptor and leads to a self-sustained pro-inflammatory deleterious cycle, contributing to an intense inflammatory process. In response to excessive ATP levels in the extracellular milieu, ATP and adenosine hydrolysis were modulated in an attempt to restrict the inflammatory process via upregulation of NTPDase and downregulation of ADA activities. We conclude that the purinergic signaling pathway modulates immune and inflammatory responses during natural infection with S. parasitica.


Assuntos
Trifosfato de Adenosina/metabolismo , Anti-Inflamatórios/metabolismo , Carpas/imunologia , Doenças dos Peixes/imunologia , Doenças dos Peixes/parasitologia , Purinérgicos/metabolismo , Transdução de Sinais , Baço/metabolismo , Adenosina Desaminase/metabolismo , Animais , Carpas/metabolismo , Modelos Animais de Doenças , Doenças dos Peixes/patologia , Proteínas de Peixes/imunologia , Micoses , Necrose , Óxido Nítrico/metabolismo , Saprolegnia/patogenicidade , Baço/patologia
5.
Fish Shellfish Immunol ; 93: 492-499, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31381973

RESUMO

TANK-binding kinase 1 (TBK1) is an important kinase that regulates the activation of interferon regulatory factor 3/7 (IRF3/7) to induce type I interferon (IFN-I) production in antiviral immune responses. However, in long-term virus-host crosstalk, viruses have evolved elaborate strategies to evade host immune defense mechanisms. In the present study, we found that grass carp (Ctenopharyngodon idella) reovirus (GCRV) hijacks TBK1 to escape IRF7-IFN-Is signaling activation. In brief, GCRV inhibited TBK1 activation by restaining K63-linked ubiquitination of TBK1 and promoting its K48-linked ubiquitination. This regulation resulted in that under low titer of GCRV infection, TBK1 overexpression specifically supressed promoter activity and phosphorylation of IRF7 and induction of downstream IFN1and IFN3. qRT-PCR data uncovered that TBK1 negatively regulated IRF7, IFN1 and IFN3 transcription levels under low viral titer infection. Along with enhancement of GCRV titers, TBK1 swiched its function to up-regulate IRF7, IFN1 and IFN3 mRNA levels. Accordingly, TBK1 promoted GCRV replication at low infected titer, but inhibited GCRV replication at high infected titer. All these results revealed a viral evasion strategy that GCRV utilizes TBK1 to block cellular IFN responses at low titers or early stages in fish species, which will lay a foundation for further researching on host-virus interactions and developing novel antiviral strategies in lower vertebrates.


Assuntos
Carpas/imunologia , Doenças dos Peixes/imunologia , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Imunidade Inata/imunologia , Animais , Fator Regulador 7 de Interferon/genética , Fator Regulador 7 de Interferon/imunologia , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/imunologia , Reoviridae/fisiologia , Infecções por Reoviridae/imunologia , Infecções por Reoviridae/veterinária
6.
Fish Shellfish Immunol ; 93: 508-516, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31352118

RESUMO

In this study, a new il-4/13 cDNA was isolated from grass carp (Ctenopharyngodon idella) using homologous cloning. The phylogenetic tree and sequence alignment of the deduced amino acid (aa) sequence showed that it was closer to grass carp il-4/13b (gcil-4/13b) than other homologues and therefore named gcil-4/13b-like (gcil-4/13bl). It has 399-nt coding sequence (CDS) which is less than gcil-4/13b (408 nt). In addition, the cloned gcil-4/13bl gene is approximately 1600 bp in length and has a conserved genetic structure consisting of four exons and three introns. Compared to gcil-4/13b gene, it has a variety of nucleotides variation across the CDS and contains a longer intron 3, suggesting that it is a new gcil-4/13 gene. The gcil-4/13bl transcripts were ubiquitously expressed in almost all selected tissues, and there was almost only gcil-4/13bl detected in brain and head kidney (HK). Recombinant grass carp (rgc) Il-4/13bl was prepared by using Escherichia coli (E. coli) Rosetta-gami 2 (DE3). The functional study demonstrated that rgcIl-4/13bl significantly upregulated arginase-2 gene expression and arginase activity, whilst downregulated nitric oxide (NO) production as well as the transcript levels of inducible nitric oxide synthesase (inos) and ifn-γ in freshly isolated grass carp HK monocytes/macrophages (M0/Mϕ). These data suggested that the newly cloned il-4/13bl had the conserved functions to activate M2-type but antagonize M1-type macrophages. Furthermore, rgcIl-4/13bl was able to drive the proliferation of M0/Mϕ which were pre-treated by rgcM-csf, indicating the involvement of gcIl-4/13bl in the proliferation of macrophages. Here we not only identified a new il-4/13-encoding gene in grass carp, but also for the first time revealed a novel function of fish Il-4/13 combined with M-csf engaging in M0/Mϕ proliferation.


Assuntos
Carpas/genética , Carpas/imunologia , Interleucina-13/genética , Interleucina-4/genética , Fator Estimulador de Colônias de Macrófagos/metabolismo , Macrófagos/imunologia , Animais , Evolução Molecular , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Interleucina-13/metabolismo , Interleucina-4/metabolismo , Macrófagos/metabolismo , Filogenia
7.
Fish Shellfish Immunol ; 93: 108-115, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31326582

RESUMO

Protein arginine methylation is a prevalent posttranslational modification and protein arginine methyltransferases 6 (PRMT6) has been identified as a suppressor of TBK1/IRF3 in human and mammals. To explore the role of PRMT6 in teleost fish, PRMT6 homologue of black carp (Mylopharyngodon piceus) has been cloned and characterized in this study. Black carp PRMT6 (bcPRMT6) transcription in host cells varies in response to different stimuli and bcPRMT6 migrates around 43 kDa in the immunoblot assay. Like its mammalian counterpart, bcPRMT6 has been identified to distribute majorly in the nucleus through the immunofluorescent staining assay. bcPRMT6 shows little interferon (IFN) promoter-inducing activity in the reporter assay and bcPRMT6 shows no antiviral activity against either grass carp reovirus (GCRV) or spring viremia of carp virus (SVCV) in plaque assay. When co-expressed with bcPRMT6, the IFN promoter-inducing abilities of black carp TBK1 (bcTBK1) and IRF3/7 (bcIRF3/7) are fiercely attenuated. Accordingly, bcTBK1-mediated antiviral activity in EPC cells is obviously dampened by bcPRMT6. The interaction between bcPRMT6 and bcIRF3/7 has been identified by co-immunoprecipitation assay; however, no direct association between bcPRMT6 and bcTBK1 has been detected. Taken together, our data elucidates for the first time in teleost fish that PRMT6 suppresses TBK1-IRF3/7 signaling during host antiviral innate immune activation.


Assuntos
Carpas/genética , Carpas/imunologia , Doenças dos Peixes/imunologia , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Sequência de Aminoácidos , Animais , Proteínas de Peixes/química , Perfilação da Expressão Gênica/veterinária , Proteínas Nucleares/química , Proteínas Nucleares/genética , Proteínas Nucleares/imunologia , Filogenia , Proteína-Arginina N-Metiltransferases/química , Proteína-Arginina N-Metiltransferases/genética , Proteína-Arginina N-Metiltransferases/imunologia , Reoviridae/fisiologia , Infecções por Reoviridae/imunologia , Infecções por Reoviridae/veterinária , Rhabdoviridae/fisiologia , Infecções por Rhabdoviridae/imunologia , Infecções por Rhabdoviridae/veterinária , Alinhamento de Sequência/veterinária , Transdução de Sinais
8.
Fish Shellfish Immunol ; 93: 278-287, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31349013

RESUMO

In order to illustrate the immunometabolic changes of fish during bacterial infection, grass carp (Ctenopharyngodon idellus) was injected with Flavobacteriumcolumnare(F.columnare) and then the immune response, nutrient metabolism and related signaling pathways were assayed from 6 h post injection (hpi) to 7 days post injection (dpi). After F.columnare injection, gill lamellae showed obvious fusion and higher mRNA expression levels of pro-inflammatory cytokines. The mRNA expression levels of TNF-α, IL-1ß and IL-8 in the head kidney were also significantly upregulated at 6 hpi and 3 dpi. Moreover, the expression of IgZ in the gill was significantly upregulated at 3 dpi and 7 dpi, while the expression of IgM in the head kidney was significantly upregulated at 1 dpi and 3 dpi after F.columnare injection. During bacterial infection, the systematic nutrient metabolism was also significantly affected. Hepatic glycolysis, indicated by GK mRNA expression and PK activity, was significantly upregulated at 1 dpi, while glucogenesis, indicated by PEPCK mRNA expression and enzyme activity, was significantly increased at later time, which resulted in the decreased hepatic glycogen content at 1dpi but increased glycogen content at 7 dpi in the experimental group. LPL, which catalyzed the lipid catabolism, showed decreased mRNA expression and enzyme activity at 6 hpi, while ACC, which was rate-limiting of FA synthesis, was significantly increased at 6 hpi, 3 dpi and 7 dpi. During this process, the nutrient sensing signaling was also significantly affected. TOR signaling in grass carp was significantly activated while ERK signaling was significantly inhibited after F.columnare infection, both of which might function as the sensor and regulator of fish immunometabolic changes.


Assuntos
Carpas/imunologia , Doenças dos Peixes/imunologia , Infecções por Flavobacteriaceae/veterinária , Imunidade Inata/genética , Nutrientes/metabolismo , Transdução de Sinais , Animais , Carpas/genética , Carpas/metabolismo , Infecções por Flavobacteriaceae/imunologia , Flavobacterium/fisiologia
9.
Fish Shellfish Immunol ; 93: 1-7, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31315061

RESUMO

Grass carp (Ctenopharyngodon idellus) is one of the most important aquaculture fish in China. This study tried to explore the effects of dietary alcoholic extract of lotus leaf (AELL) addition on the growth performance and health status of grass carp by feeding juvenile fish (average weight: 34 ±â€¯1 g) with four different experimental diets: control, AELL7, AELL14 and AELL21 for 8 weeks. At the end of the growth trial, the highest values of final body weight (FBW), weight gain rate (WGR), specific growth rate (SGR) and feed intake (FI) all occurred in group AELL14 (P < 0.05). Compared to control, the crude lipid content of whole-body and the serum malondialdehyde (MDA) in the three experimental groups decreased, while the serum superoxide dismutase (SOD), glutathione peroxidase (GSH) and total antioxidant capacity (T-AOC) values almost all increased in the three experimental groups. The highest serum immunoglobulin M (IgM) concentration occurred in AELL14 group (P < 0.05). In AELL14 and AELL21 groups, both the serum complement 3 (C3) concentration and lysozyme (LYS) activity were significantly higher, whereas the final cumulative mortality in challenge test was significantly lower, when compared to those in control group (P < 0.05). The AELL exerted dose-dependent beneficial effects on grass carp health through up-regulating related gene expressions and enzyme activity. In conclusion, the optimal dietary AELL level is 0.14% for juvenile grass carp.


Assuntos
Carpas/imunologia , Nelumbo/química , Extratos Vegetais/metabolismo , Ração Animal/análise , Animais , Análise Química do Sangue/veterinária , Carpas/crescimento & desenvolvimento , Carpas/metabolismo , Dieta/veterinária , Suplementos Nutricionais/análise , Relação Dose-Resposta a Droga , Feminino , Masculino , Extratos Vegetais/administração & dosagem , Distribuição Aleatória
10.
Microbiol Immunol ; 63(9): 379-391, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31310013

RESUMO

The immune system with large number of molecules protects the host against a plethora of continuously evolving microbes. Major histocompatibility complex (MHC) molecules serve as cardinal elements of the adaptive immune system responsible for the activation of the adaptive immunity in the host. The present study reports MHCI molecule in freshwater carp, Catla catla, and its differential expression in immunologically relevant tissues post-infection with Gram-negative and Gram-positive bacteria. The MHCI sequence of C. catla had 502 bp nucleotides encoding putative 146 amino acids. The phylogenetic analysis exhibited its evolutionary conservation within the Cyprinidae family and formed a different clade with the higher vertebrates. Simultaneously, CXCR3 and CXCR4 chemokines were cloned and characterized for their expression in infected tissues. Analysis of immunologically relevant tissues of the infected fish exhibited an increase of MHCI gene expression and the down-regulation of CXCR3 and CXCR4 chemokines, indicating a tricky interaction between the innate and adaptive immune system. It was found that intestine, skin and spleen played a crucial role in the contribution of the defense activity which instigated the self-immunity. These immune activities can provide useful information to understand the interaction of self and non-self- immune system in freshwater fish, Catla catla.


Assuntos
Carpas/genética , Quimiocinas/genética , Clonagem Molecular/métodos , Cyprinidae/genética , Genes MHC Classe I/genética , Receptores CXCR3/genética , Receptores CXCR4/genética , Imunidade Adaptativa/genética , Sequência de Aminoácidos , Animais , Carpas/imunologia , Cyprinidae/imunologia , Regulação para Baixo , Doenças dos Peixes/imunologia , Doenças dos Peixes/microbiologia , Água Doce , Expressão Gênica , Filogenia , Alinhamento de Sequência , Pele/imunologia , Baço/imunologia
11.
Fish Shellfish Immunol ; 93: 1093-1099, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31310849

RESUMO

Chlorpyrifos (CPF) has become a mainly pollution in water environment. Micro-RNAs (miRNAs) play an important part in the development of apoptosis and autophagy. However, the potential mechanism of CPF induced kidney toxicity and the roles of miRNAs are still unclear. To explore the underlying mechanism, the kidney of common carp exposed to different concentrations of CPF for 40 days was used as a research object. We found that CPF could damage the ultrastructure and function of kidney; and also caused antioxidant system disorder. CPF inhibited the mRNA level of miR-19a which improved AMP-activated protein kinase (AMPK). Furthermore, the detection of apoptosis and autophagy relative genes showed that the expressions of TSC complex subunit 2 (TSC2), light chain 3 (LC3), Dynein, tumor protein 53 (p53), Bcl-2 associated X protein (Bax), caspase-3 and caspase-9 were enhanced and the expressions of nechanistic target of rapamycin (mTOR), Ras homolog mTORC1 binding (Rheb) and B-cell lymphoma (Bcl-2) were reduced in dose-dependent way. Taken together, we conclude that CPF causes oxidative stress and miR-19a-AMPK axis disorder, thereby promotes apoptosis and autophagy in common carp kidney. Our study will provide theoretical basis for toxicology research and environmental protection of CPF.


Assuntos
Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Carpas/fisiologia , Clorpirifos/efeitos adversos , Proteínas de Peixes/genética , Regulação da Expressão Gênica/imunologia , Poluentes Químicos da Água/efeitos adversos , Proteínas Quinases Ativadas por AMP/genética , Proteínas Quinases Ativadas por AMP/metabolismo , Animais , Carpas/genética , Carpas/imunologia , Relação Dose-Resposta a Droga , Proteínas de Peixes/metabolismo , Inseticidas/efeitos adversos , Rim/efeitos dos fármacos , Rim/enzimologia , MicroRNAs/genética , MicroRNAs/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Distribuição Aleatória
12.
Fish Shellfish Immunol ; 92: 706-711, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31276789

RESUMO

Recently, studies have shown that IκB kinase ß (IKKß), a critical kinase in the nucleus factor kappa-B (NF-κB) pathway, participates in inflammatory responses associated with unfolded protein response (UPR) and plays an important role in ER stress-induced cell death. The unfolded protein response (UPR), which is a regulatory system to restore cellular homeostasis in the endoplasmic reticulum (ER), such as oxidative stress, bacterial infection, and virus invasion. The UPR pathways have been reported to be involved in immune responses in mammals, including the classical NF-κB pathway. However, the molecular mechanism of their crosstalk remains to be elucidated. Previously, we demonstrated that IKKß also has some conserved functions between fish and human, as grass carp (Ctenopharyngodon idella) IKKß (CiIKKß) can activate NF-κB pathway. In this study, we found that CiIKKß level in nucleus was elevated under ER stress and CiIKKß can interact with grass carp X-box-binding protein 1 (CiXBP1S), a key transcription factor in UPR. Consistently, fluorescent histochemical analysis of grass carp kidney (CIK) cells indicated that CiIKKß and CiXBP1S colocalized under ER stress. Furthermore, overexpression of CiIKKß in CIK cells enhanced ER stress tolerance by regulating UPR signaling and resulted in the significant increase of cell viability.


Assuntos
Carpas/genética , Estresse do Retículo Endoplasmático , Proteínas de Peixes/genética , Regulação da Expressão Gênica/imunologia , Quinase I-kappa B/genética , Proteína 1 de Ligação a X-Box/genética , Animais , Carpas/imunologia , Núcleo Celular/genética , Sobrevivência Celular , Proteínas de Peixes/imunologia , Quinase I-kappa B/imunologia , Resposta a Proteínas não Dobradas , Proteína 1 de Ligação a X-Box/imunologia
13.
Fish Shellfish Immunol ; 92: 612-620, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31265909

RESUMO

Present study evaluated the effects of curcumin, the principal curcuminoid of turmeric, on Cyprinus carpio growth, skin mucosal immune parameters, immune-related gene expression, and susceptibility to pathogen challenge. Diets containing four various concentrations of curcumin (g Kg-1): 0 g [basal diet], 5 g [T5], 10 g [T10], and 15 g [T15] were fed to the carp (average weight: 16.37 g) for 8 weeks. Growth parameters were analysed at 4 and 8 weeks post-feeding. Skin mucosal immune responses and expression were examined in 8 weeks post-feeding. Growth performance was significantly higher in T10 and T15, with final weight gain of 102.26 ±â€¯2.31 g and specific growth rate of 3.24 ±â€¯0.37, respectively. The lowest feed conversion ratio (2.35 ±â€¯0.16) was recorded in T15 than in the control (P < 0.05). Among the skin mucosal immune parameters examined, lysozyme (36.8 ±â€¯4.03 U mL-1), total immunoglobulin (6.74 ±â€¯0.5 mg mL-1), protein level (18.7 ±â€¯1.62 mg mL-1), alkaline phosphatase (96.37 ±â€¯6.3 IU L-1), and protease activity (9.47 ±â€¯0.82%) were significantly higher in T15, while the peroxidase activity was higher in T10 (10.24 ±â€¯0.9 U mg-1 protein). Further, lysozyme, superoxide dismutase (SOD) and catalase (CAT) activities were measured in serum and found to be higher in T10 or T15 than in the control (P < 0.05). However, malondialdehyde level decreased significantly in T10 and T15. Furthermore, antioxidant genes (SOD, CAT, nuclear factor erythroid 2-related factor 2) and anti-inflammatory cytokine Interleukin-10 were upregulated in the head kidney, intestine, and hepatopancreas of fish in T10 and T15. Conversely, expression of pro-inflammatory cytokines (IL-1ß, tumour necrosis factor-alpha), signalling molecule NF-κBp65 were down-regulated in the tested tissues of T10 and T15. Expression of Toll-like receptor 22 (TLR22) was down regulated in head-kidney and intestine of T15. Fish from T15 exhibited significantly higher relative post-challenge survival (69.70%) against Aeromonas hydrophila challenge. Results of the present study suggest that dietary supplements of curcumin at 15 g Kg-1can significantly improve the growth performance, skin mucosal and serum antioxidant parameters, and strengthen the immunity of C. carpio. Therefore, curcumin represents a promising food additive for carps in aquaculture.


Assuntos
Antioxidantes/metabolismo , Carpas/imunologia , Curcuma/química , Resistência à Doença , Doenças dos Peixes/imunologia , Expressão Gênica/efeitos dos fármacos , Imunidade nas Mucosas/efeitos dos fármacos , Aeromonas hydrophila/fisiologia , Ração Animal/análise , Animais , Carpas/crescimento & desenvolvimento , Citocinas/genética , Citocinas/metabolismo , Dieta/veterinária , Suplementos Nutricionais/análise , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/veterinária , Compostos Fitoquímicos/farmacologia , Distribuição Aleatória , Pele/imunologia
14.
Fish Shellfish Immunol ; 93: 73-81, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31302283

RESUMO

In the present study, we reported 18 LAB strains isolated from the intestinal contents of Cyprinus carpio, and their probiotic properties both in vitro and in vivo. The results showed that 9 of them had higher in vitro immunomodulatory properties, effectively survived under acidic (pH 2.5) and bile salt (ranging from 0.1% to 0.5%) conditions, and inhibited the growth of 4 pathogens. Among them, Lactococcus lactis Q-8, Lactococcus lactis Q-9, and Lactococcus lactis Z-2 showed the strongest adhesion abilities and inhibition of pathogen adhesion to mucin. When the fish consumed diets containing these 3 strains (5 × 108 CFU/g) for 8 weeks, the weight gain (WG) and specific growth rate (SGR) had significantly (P < 0.05) increased, especially with L. lactis Q-8, which had a WG of 231.45%, and SGR of 2.22%. Survival rate in each LAB supplementation group was also significantly higher than that in control group during the feeding period (P < 0.05). For the cytokines expression levels in serum, different expression patterns were also observed. Before the infection with Aeromonas hydrophila, L. lactis supplementation significant up-regulated protein levels of pro-inflammatory cytokines (TNF-α, IL-1ß, IL-6, IL-12) compared with negative (CK1) group, while these cytokines were significantly lower than those in positive (CK2) group after infection. However, whether infected or not, the expression of anti-inflammatory cytokines (IL-10, TGF-ß) were significantly increased in L. lactis Q-8, L. lactis Q-9, and L. lactis Z-2 treatment groups. In conclusion, these 3 L. lactis strains screened from common carp were effective in improving growth, innate immunity and disease resistance. Based on the physiological characteristics in our study, they might be used as potential probiotics in aquaculture.


Assuntos
Carpas/imunologia , Resistência à Doença/efeitos dos fármacos , Doenças dos Peixes/imunologia , Imunidade Inata/efeitos dos fármacos , Lactococcus lactis/química , Probióticos/farmacologia , Aeromonas hydrophila/fisiologia , Ração Animal/análise , Animais , Carpas/crescimento & desenvolvimento , Dieta/veterinária , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/veterinária
15.
Fish Shellfish Immunol ; 92: 315-321, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31202965

RESUMO

Mammalian Interleukin (IL)-23 is a heterodimeric cytokine with an IL-23-specific P19 subunit and a P40 subunit shared with IL-12, and plays a key role in the regulation of cell differentiation as well as inflammation. We previously demonstrated the existence of three soluble fish Interleukin (Il)-23 isoforms consist of a single P19 and one of three P40 isoforms (P40a/b/c) in grass carp. In the present study, three recombinant grass carp Il-23 (rgcIl-23) isoforms were prepared by linking gcP19 and gcP40a/b/c in a prokaryotic expression system, and then their functional properties were verified in grass carp head kidney leukocytes (HKLs). All three rgcIl-23 isoforms showed the bioactivities to divergently upregulate the mRNA expression of Th17 signature cytokines (il17a/f1, il21, il22 and il26) as well as Il-23 receptor (il23r) in HKLs. Moreover, they also promoted gcIl-17a/f1 secretion in a dose-dependent manner, strengthening their roles in Th17-like response. Furthermore, induction of il17a/f1 and il23r transcription by rgcIl-23 was blocked by a STAT3 inhibitor in grass carp HKLs, suggesting the involvement of STAT3 signaling in these inductions. Taken together, we for the first time identified the bioactivities of fish Il-23 isoforms and particularly revealed the existence of Il-23/Il-17a/f1 axis in fish, thereby advancing our understanding of Th17-like responses in fish immunity.


Assuntos
Carpas/genética , Carpas/imunologia , Proteínas de Peixes/genética , Interleucina-23/genética , Células Th17/imunologia , Animais , Citocinas/genética , Citocinas/imunologia , Proteínas de Peixes/metabolismo , Regulação da Expressão Gênica , Rim Cefálico/imunologia , Interleucina-23/metabolismo , Leucócitos/imunologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Transdução de Sinais/imunologia
16.
Fish Shellfish Immunol ; 92: 288-299, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31195114

RESUMO

Omnivorous fish species such as the common carp (Cyprinus carpio) are able to biosynthesise long chain polyunsaturated fatty acids (LC-PUFAs) from plant oil PUFA precursors, but the influence of the amount and quality of the LC-PUFAs biosynthesised from these oils on the immunocompetence status of the fish has received little attention. This study aims to evaluate whether the conversion of PUFA by carp induces a sufficient biosynthesis of LC-PUFA to maintain a good immunocompetence status in this species. Six iso-nitrogenous (crude protein = 39.1%) and iso-lipidic (crude lipids = 10%) diets containing three different lipid sources (cod liver oil (CLO) as fish oil; linseed oil (LO) and sunflower oil (SFO) as plant oils) were formulated with or without ß-glucan supplementation at 0.25 g/kg diet. Juvenile carp (16.3 ±â€¯0.6 g initial body weight) were fed a daily ration of 4% body weight for 9 weeks and then infected at day 64 with the bacteria Aeromonas hydrophyla. No significant differences in survival rate, final body weight, specific growth rate and feed conversion rate were observed between diets. After bacterial infection, mortality rate did not differ between fish fed CLO and plant oil-based diets, indicating that the latter oils did not affect the overall immunocompetence status of common carp. Plant oil-based diets did not alter lysozyme activity in healthy and infected fish. No negative effects of plant oils on complement activity (ACH50) were observed in healthy fish, even if both plant oil-based diets induced a decrease in stimulated fish two days after infection. Furthermore, the levels of various immune genes (nk, lys, il-8, pla, pge, alox) were not affected by plant oil-based diets. The expression of pla and pge genes were higher in SFO-fed fish than in CLO ones, indicating that this plant oil rich in linoleic acid (LA) better stimulated the eicosanoid metabolism process than fish oil. In response to ß-glucan supplementation, some innate immune functions seemed differentially affected by plant oil-based diets. LO and SFO induced substantial LC-PUFA production, even if fish fed CLO displayed the highest EPA and DHA levels in tissues. SFO rich in LA induced the highest ARA levels in fish muscle while LO rich in α-linolenic acid (ALA) sustained higher EPA production than SFO. A significantly higher fads-6a expression level was observed in SFO fish than in LO ones, but this was not observed for elovl5 expression. In conclusion, the results show that common carp fed plant oil-based diets are able to produce substantial amounts of LC-PUFA for sustaining growth rate, immune status and disease resistance similar to fish fed a fish oil-based diet. The differences in the production capacity of LC-PUFAs by the two plant oil-based diets were associated to a differential activation of some immune pathways, explaining how the use of these oils did not affect the overall immunocompetence of fish challenged with bacterial infection. Moreover, plant oil-based diets did not induce substantial negative effects on the immunomodulatory action of ß-glucans, confirming that these oils are suitable for sustaining a good immunocompetence status in common carp.


Assuntos
Carpas/imunologia , Imunocompetência/efeitos dos fármacos , Óleos Vegetais/metabolismo , beta-Glucanas/metabolismo , Ração Animal/análise , Animais , Carpas/crescimento & desenvolvimento , Dieta/veterinária , Suplementos Nutricionais/análise , Ácidos Graxos Insaturados/metabolismo , Óleos de Peixe/administração & dosagem , Proteínas de Peixes/imunologia , Proteínas de Peixes/metabolismo , Óleos Vegetais/administração & dosagem , Distribuição Aleatória , beta-Glucanas/administração & dosagem
17.
Environ Pollut ; 246: 963-971, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31159146

RESUMO

Bisphenol S (BPS) has been widely used as a bisphenol alternative in recent few years. However, with mounting evidence suggesting that the presence of BPS in the environment also poses risks to ecosystems and human health, we decided to use the juvenile common carp (Cyprinus carpio) and its primary macrophages as in vivo and in vitro models to examine if BPS is a safe substitute of BPA. The present study evaluated the immune responses of chronic BPS exposure and their mechanisms of action associated with peroxisome proliferator-activated receptor (PPAR) signaling pathway. Potential oxidative stress and pro-inflammatory effects of BPS exposure were identified in fish liver after 60-day exposure, based on the increased reactive oxygen species (ROS) production, antioxidant capacity, NO production, lipid peroxidation, and induction of inflammatory cytokine expression, as well as acute phase protein levels of C-reactive protein, immunoglobulin M, lysozyme, and complement component 3. Moreover, pparγ, PPAR pathway-associated genes retinoid x receptor α (rxrα) and nuclear factor-κb (nfκb) presented a rough concentration-dependent alteration after BPS exposure. An acute BPS exposure to the isolated primary macrophages from juvenile common carp was performed to help elucidate gene expression patterns of pparγ, rxrα, and nfκb in a typical immune cell model, the results were consistent with what we found in vivo experiments for long-term BPS exposure. Furthermore, with coexposure to BPS and a PPARγ antagonist, the restriction of PPAR signaling pathway significantly inhibited the induction of ROS and the mRNA level of interleukin-1ß, confirming the involvement of PPAR pathway in BPS-induced chronic inflammatory stress in liver.


Assuntos
Poluentes Ambientais/toxicidade , Inflamação/metabolismo , Fígado/efeitos dos fármacos , PPAR gama/metabolismo , Fenóis/toxicidade , Sulfonas/toxicidade , Animais , Carpas/imunologia , Carpas/metabolismo , Inflamação/genética , Fígado/imunologia , Fígado/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Macrófagos/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/imunologia , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética
18.
Fish Shellfish Immunol ; 92: 331-340, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31176765

RESUMO

Red blood cells (RBCs) are widely accepted as their primary function in respiration. Recent studies in mammals have revealed a vital role in immune responses of RBCs; however, little is known about immune function of teleost erythrocytes. Here we demonstrated that RBCs from grass carp (Ctenopharyngodon idella) were capable of binding and aggregating the bacteria with apparent morphological alterations. The phagocytosis by teleost RBCs (erythrophagocytosis) was visualized by confocal, scanning and transmission electron microscopy. Hb-FeII of hemoglobin (Hb) could quickly be auto-oxidated to Hb-FeIII (methemoglobin/metHb) in the presence of oxygen (O2), and release superoxide radical (O2-.) which could be spontaneously dismutated into H2O2 that could further oxidize Hb-FeIII to transient HbFeIV-OH (ferryl-Hb). Furthermore, bacterial extracellular proteases and pathogen-associated molecular patterns (PAMPs) binding to Hb could synergistically activate pseudoperoxidase, subsequently facilitated the generation of reactive oxygen species (ROS) which were toxic to the bacteria. Our results indicated that erythrocyte pertains anti-bacterial activity using unique ROS generation pathway via oxidation of hemoglobin and associated with its phagocytosis.


Assuntos
Antibacterianos/imunologia , Carpas/imunologia , Eritrócitos/imunologia , Fagocitose/imunologia , Espécies Reativas de Oxigênio/imunologia , Aeromonas hydrophila/efeitos dos fármacos , Animais , Escherichia coli/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos
19.
Fish Shellfish Immunol ; 92: 552-569, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31252043

RESUMO

This study investigated the effects of bile acid (BA) supplementation on growth performance, intestinal immune function and the mRNA expression of the related signalling molecules in on-growing grass carp (Ctenopharyngodon idella). A total of 540 healthy grass carp (mean weight 179.85 ±â€¯1.34 g) were fed a normal protein and lipid (NPNL) diet containing 29% crude protein (CP) and 5% ether extract (EE), and five low-protein and high-lipid (LPHL) diets (26% CP, 6% EE) with graded levels of BA (0-320 mg/kg diet) for 50 days. The fish were then challenged with Aeromonas hydrophila for 14 days. The results indicated that compared with the NPNL diet, the LPHL diet (unsupplemented BA) suppressed the growth performance, intestinal development and enteritis resistance capability and impaired the partial intestinal immune function of on-growing grass carp. Whereas in the LPHL diet, optimal BA supplementation significantly improved fish growth performance (percent weight gain, specific growth rate, feed intake and feed efficiency) and intestinal growth and function (intestine weight, intestine length and intestosomatic index), increased beneficial bacteria Lactobacillus and Bifidobacterium amounts, decreased harmful bacteria Aeromonas and Escherichia coli amounts, elevated lysozyme and acid phosphatase activities, increased complement (C3 and C4) and immunoglobulin M contents, and upregulated ß-defensin-1, hepcidin, liver expressed antimicrobial peptide 2A (LEAP-2A), LEAP-2B, Mucin2, interleukin 10 (IL-10), IL-11, transforming growth factor (TGF)-ß1, TGF-ß2, IL-4/13A (not IL-4/13B), TOR, S6K1 and inhibitor of κBα (IκBα) mRNA levels. In addition, optimal BA supplementation in the LPHL diet downregulated tumour necrosis factor α (TNF-α), interferon γ2 (IFN-γ2), IL-1ß, IL-6, IL-8, IL-15, IL-17D, IL-12p35, IL-12p40 (rather than proximal intestine (PI) or mid intestine (MI), nuclear factor kappa B p65 (NF-κB p65) (except NF-κB p52), c-Rel, IκB kinase ß (IKKß), IKKγ (except IKKα), eIF4E-binding proteins (4E-BP)1 and 4E-BP2 mRNA levels in all three intestinal segments of on-growing grass carp (P < 0.05). These findings suggest that BA supplementation in the LPHL diet improves growth and intestinal immune function of fish. Furthermore, 240 mg/kg BA supplementation in the LPHL diet was superior to the NPNL diet in improving growth and enhancing intestinal immune function of fish. Finally, based on percent weight gain, feed intake, protecting fish against enteritis, lysozyme activity in MI and acid phosphatase activity in distal intestine (DI), the optimal BA supplementation for on-growing grass carp were estimated to be 168.98, 170.23, 166.67, 176.50 and 191.97 mg/kg diet, respectively.


Assuntos
Ácidos e Sais Biliares/metabolismo , Carpas/imunologia , Metabolismo dos Lipídeos , NF-kappa B/genética , Serina-Treonina Quinases TOR/genética , Ração Animal/análise , Animais , Ácidos e Sais Biliares/administração & dosagem , Carpas/genética , Carpas/crescimento & desenvolvimento , Carpas/metabolismo , Dieta/veterinária , Suplementos Nutricionais/análise , Expressão Gênica/imunologia , Intestinos/imunologia , NF-kappa B/imunologia , RNA Mensageiro/genética , Transdução de Sinais , Serina-Treonina Quinases TOR/imunologia
20.
Fish Shellfish Immunol ; 92: 590-599, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31252044

RESUMO

HIFs (Hypoxia inducible factors) are the main regulators of the expression change of oxygen-dependent genes, in addition, they also play important roles in immune regulation. HIFs participate in infectious diseases and inflammatory responses, providing us a new therapeutic target for the treatment of diseases. In this study, 16 HIFs were identified in common carp genome database. Comparative genomics analysis showed large expansion of HIF gene family and approved the four round whole genome duplication (WGD) event in common carp. To further understand the function of HIFs, the domain architectures were predicted. All HIF proteins had the conserved HLH-PAS domain, which were essential for them to form dimer and bind to the downstream targets. The differences in domain of HIFα and HIFß might result in their different functions. Phylogenetic analysis revealed that all HIFs were divided into two subfamilies and the HIFs in common carp were clustered with their teleost counterparts indicating they are highly conservative during evolution. In addition, the tissue distribution was examined by RT-PCR showed that most of HIF genes had a wide range of tissue distribution but exhibited tissue-specific expression patterns. The expression divergences were observed between the copy genes, for example, HIF1A-1, HIF2A-1, ARNT-2 had wide tissue distribution while their copies had limited tissue distribution, proving the function divergence of copies post the WGD event. In order to find an effective activation of HIFs and apply to treatment of aquatic diseases, we investigate the dietary supplementation effects of different strains of Lactococcus lactis on the expression of HIFα subfamily members in kidney of common carp infected with A. hydrophila. In addition, all of the HIF genes have a high expression in the early stages of infection, and decreased in the treatment time point of 48 h in common carp. This phenomenon confirms that as a switch, the main function of HIFs is to regulate the production of immune response factors in early infection. So activation of the switch may be an effective method for infectious disease treatment. As expected, the treatment groups improved the expression of HIFs compared with the control group, and the effects of the three strains are different. The strain1 of L. lactis had a stronger induction on HIF genes than strain2 and strain3, and it might be applied as a potential activation of HIF genes for disease treatment. So, adding befitting L. lactis maybe a well method to activate the HIF genes to protect them from mycobacterial infection.


Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Carpas/genética , Carpas/imunologia , Doenças dos Peixes/imunologia , Expressão Gênica , Lactococcus lactis/química , Probióticos/metabolismo , Aeromonas hydrophila/fisiologia , Ração Animal/análise , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Dieta/veterinária , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/veterinária , Família Multigênica , Probióticos/administração & dosagem
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