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1.
Mol Immunol ; 124: 172-179, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32585511

RESUMO

The family of TRIM proteins with E3 ubiquitin ligase activity plays important roles in virus infection in vertebrates and invertebrates. In this study, a novel Trim gene shows high similarity to Trim23 (designated as MnTrim23) was identified from Macrobrachium nipponense. The MnTrim23 protein contains three conserved domains (one RING finger domain, two B-box, and one Coiled-coil region) at its N-terminal and one ARF domain at its C-terminal. The ARF domain characterizes the members of the Trim23 family. MnTrim23 belongs to C-IX family. Phylogenetic analysis shows that MnTrim23 has a closer genetic distance with other Trim23 proteins from invertebrates than that from vertebrates. MnTrim23 has higher expression level in the intestine and hepatopancreas than in the other immune tissues. The expression levels of MnTrim23 in the gills, stomach, and intestines are significantly up-regulated after white spot syndrome virus (WSSV) infection. Moreover, knockdown of MnTrim23 inhibits WSSV replication and VP28 expression, suggesting that MnTrim23 plays a positive role in WSSV infection. Further studies revealed that MnTrim23 negatively regulates the Relish transcription factor-mediated expression of antimicrobial peptides (AMPs). Synthetic AMPs inhibit VP28 expression and WSSV replication. These findings indicate that Trim23 promotes WSSV replication by inhibiting the expression of AMPs that are positively regulated by the host NF-κB signal pathway.


Assuntos
Peptídeos Catiônicos Antimicrobianos/biossíntese , Interações entre Hospedeiro e Microrganismos/fisiologia , Penaeidae/virologia , Proteínas com Motivo Tripartido/metabolismo , Vírus da Síndrome da Mancha Branca 1/fisiologia , Animais , Peptídeos Catiônicos Antimicrobianos/imunologia , Regulação da Expressão Gênica/fisiologia , Penaeidae/imunologia , Penaeidae/metabolismo , Proteínas com Motivo Tripartido/imunologia , Replicação Viral/fisiologia
2.
Food Chem ; 324: 126888, 2020 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-32344352

RESUMO

This study aimed to investigate if myo-inositol (MI) supplementation could alleviate adverse effects caused by aflatoxin B1 (AFB1) with respect to growth performance, AFB1 residues, immune response and antioxidant status of Litopenaeus vannamei. 800 shrimp (initial weight: 1.1 g) were divided into five groups: MI0 (basal diet); MI0 + LA, MI0 + HA, MI200 + LA and MI200 + HA fed with AFB1-contaminated diets (LA, low concentration AFB1; HA, high concentration AFB1; MI200, adding 200 mg MI kg-1 diet). The results showed that HA significantly decreased growth performance, systemic inositol content and lipid content. AFB1 residues were detected in the hepatopancreas of shrimp, but not the muscle. Histological lesions were observed in MI0 + LA and MI0 + HA groups. HA supplementation raised malondialdehyde and protein carbonyl content and reduced some antioxidant enzyme activities and immune-related genes expression, which was slightly ameliorated by MI supplementation. Our results suggest that myo-inositol may slightly mitigate negative impacts caused by AFB1 in L. vannamei.


Assuntos
Aflatoxina B1/análise , Antioxidantes/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Inositol/farmacologia , Penaeidae/crescimento & desenvolvimento , Aflatoxina B1/administração & dosagem , Alanina Transaminase/metabolismo , Animais , Aspartato Aminotransferases/metabolismo , Dieta , Suplementos Nutricionais , Hepatopâncreas/enzimologia , Hepatopâncreas/metabolismo , Malondialdeído/metabolismo , Penaeidae/imunologia , Penaeidae/metabolismo , Carbonilação Proteica/efeitos dos fármacos
3.
Mol Immunol ; 120: 113-121, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32113131

RESUMO

Litopenaeus vannamei, as an important marine aquaculture species, has attracted more and more attentions in past several years. More recently people got its genome fine mapping, which unveiled a gene treasure. In this study, we have identified a novel trypsin-like protein which came from previous WSSV-infected shrimp plasma iTRAQ data. This protein is a 39 kDa protein with 363 amino acids. It contains a conserved trypsin-domain and could be strongly induced with WSSV infection. Interestingly, knockdown of this protein made shrimps vulnerable to WSSV infection. Further exploration unveiled that this fragility was probably due to the fact that knockdown of this protein could cause shrimp hemocytes apoptosis, which indicated that this protein played key roles in preventing shrimp hemocytes from apoptosis. To further explore how LvTLAP protected shrimp hemocytes from apoptosis, GST pull down assay was applied to screen LvTLAP interacting protein in shrimp plasma. L. vannamei growth and transformation-dependent-like protein (LvGTD-like protein) was identified as a LvTLAP interacting protein, which played proapoptotic roles in cells. Thus, a possible explanation for LvTLAP anti-apoptosis activity was that this protein could block LvGTD-like protein proapoptotic activity to protect shrimp hemocytes from death. In general, our study has uncovered a novel WSSV responsive shrimp plasma protein, which played key roles in shrimp hemocytes anti-apoptosis and shrimp against WSSV infection.


Assuntos
Proteínas Sanguíneas/imunologia , Penaeidae/imunologia , Penaeidae/virologia , Vírus da Síndrome da Mancha Branca 1/patogenicidade , Sequência de Aminoácidos , Animais , Apoptose/genética , Apoptose/imunologia , Apoptose/fisiologia , Sequência de Bases , Proteínas Sanguíneas/química , Proteínas Sanguíneas/genética , DNA/genética , Técnicas de Silenciamento de Genes , Hemócitos/metabolismo , Hemócitos/patologia , Hemócitos/virologia , Penaeidae/genética
4.
Fish Shellfish Immunol ; 96: 319-329, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31805414

RESUMO

Viral immediate early (IE) genes encode regulatory proteins that are critical for viral replication. WSV056 is an IE protein of white spot syndrome virus (WSSV), an important pathogen of farmed shrimp. It targets the host Rb protein(s) and, according to a previous study, may enhance the replication of the viral genome. However, the ectopic expression of WSV056 in transgenic Drosophila melanogaster exerted an inhibitory effect on the replication of Drosophila C virus (DCV). Transcriptome study using Affymetrix GeneChip suggested that the enrichment of serine proteases (SPs) likely accounts for DCV inhibition in WSV056-overexpressing Drosophila. Injection of recombinant WSV056 to the WSSV natural host Litopenaeus vannamei enhanced the expression of the SP family member prophenoloxidase-activating enzyme 2 (LvPPAE2) and conferred shrimp with more resistance to WSSV infection. LvPPAE2 knockdown contributed to decreased expression of antimicrobial peptides LvAlf1 and LvLyz1, reduced hemolymph phenoloxidase activity, and increased virus load, suggesting that LvPPAE2 is involved in the host defense against WSSV infection. Taken together, these results suggest that wsv056 plays a role in restricting viral replication by inducing the SP-mediated immune responses in the host.


Assuntos
Imunidade Inata/genética , Penaeidae/genética , Penaeidae/imunologia , Serina Endopeptidases/genética , Serina Endopeptidases/imunologia , Vírus da Síndrome da Mancha Branca 1/fisiologia , Animais , Animais Geneticamente Modificados/genética , Animais Geneticamente Modificados/imunologia , Peptídeos Catiônicos Antimicrobianos/genética , Peptídeos Catiônicos Antimicrobianos/metabolismo , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/imunologia , Drosophila melanogaster/genética , Drosophila melanogaster/imunologia , Análise Serial de Proteínas
5.
Fish Shellfish Immunol ; 96: 53-61, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31801694

RESUMO

Target of rapamycin (TOR) is an atypical of Ser/Thr protein kinase that plays an important role in many aspects such as cell growth, reproduction, differentiation, cell cycle regulation, autophagy and apoptosis. However, little information is known about the enzyme in crustaceans. Here, a novel TOR was identified from shrimp Penaeus vannamei (PvTOR) and its biological functions were investigated in response low temperature stress. The PvTOR gene encoded a polypeptide of 2464 amino acids with an estimated molecular mass of 279.4 kD and a predicted isoelectronic point (pI) of 7.30. Phylogenetic analysis revealed that PvTOR shared high similarity with other known species. PvTOR mRNA was detected in all the tested tissues and highest transcription in muscle and hepatopancreas. PvTOR transcriptional level was up-regulated significantly at 1.5 h and 3 h, and down-regulated at 12 h and 24 h after low temperature stress. TEM and autophagy indicator system GFP-PvLC3 suggested that low temperature induced autophagy generation. ROS, Ca2+ concentration and apoptosis rate were increased significantly in TOR-knockdown shrimp after low temperature stress. The autophagy associated gene ATG8II/I, PvBeclin-1, PvATG14, apoptosis gene PvPARP, Pvcasp-3, PvBAX and Pvp53 transcripts, and casp-3/8 activity in hemocyte were increased significantly in TOR-knockdown group shrimp at 3 h after low temperature stress. Additionally, THC counts of TOR-knockdown group were significantly higher than the dsGFP group. In summary, these results suggested that PvTOR plays an important role in the adaptation mechanisms of shrimp at low temperature by regulating autophagy and apoptosis.


Assuntos
Proteínas de Artrópodes/genética , Temperatura Baixa/efeitos adversos , Penaeidae/genética , Penaeidae/imunologia , Transdução de Sinais/genética , Serina-Treonina Quinases TOR/genética , Animais , Apoptose/genética , Proteínas de Artrópodes/metabolismo , Autofagia/genética , Filogenia , Análise de Sequência de DNA , Estresse Fisiológico , Serina-Treonina Quinases TOR/metabolismo
6.
Fish Shellfish Immunol ; 96: 245-253, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31830564

RESUMO

RNA polymerase (RNAP) II (DNA-directed) (POLR2) genes are essential for cell viability under environmental stress and for the transfer of biological information from DNA to RNA. However, the function and characteristics of POLR2 genes in crustaceans are still unknown. In the present study, a POLR2H cDNA was isolated from Pacific white shrimp (Litopenaeus vannamei) and designated as Lv-POLR2H. The full-length Lv-POLR2H cDNA is 772 bp in length and contains a 32-bp 5'- untranslated region (UTR), a 284-bp 3'- UTR with a poly (A) sequence, and an open reading frame (ORF) of 456 bp encoding an Lv-POLR2H protein of 151 amino acids with a deduced molecular weight of 17.21 kDa. The Lv-POLR2H protein only contains one functional domain, harbors no transmembrane domains and mainly locates in the nucleus. The expression of the Lv-POLR2H mRNA was ubiquitously detected in all selected tissues, with the highest level in the gills. In situ hybridization (ISH) analysis showed that Lv-POLR2H was mainly located in the secondary gill filaments, the transcript levels of Lv-POLR2H in the gills were found to be significantly affected after challenge by pH, low salinity and high concentrations of NO2- and NH4+, indicating that Lv-POLR2H in gill tissues might play roles under various physical stresses. Specifically, under high-pH stress, knockdown of Lv-POLR2H via siRNA significantly decreased the survival rate of the shrimp, indicating its key roles in the response to high-pH stress. Our study may provide the first evidence of the role of POLR2H in shrimp responding to high-pH stress and provides new insight into molecular regulation in response to high pH in crustaceans.


Assuntos
Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Penaeidae/genética , Penaeidae/imunologia , Peptídeos/genética , Peptídeos/imunologia , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/imunologia , Sequência de Bases , Perfilação da Expressão Gênica , Brânquias/metabolismo , Concentração de Íons de Hidrogênio , Peptídeos/química , Filogenia , Estresse Fisiológico
7.
J Immunoassay Immunochem ; 41(1): 45-59, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31630605

RESUMO

The present study investigated the effects of short-chain fructooligosaccharides (FOS) as dietary supplements on stress, immune response, gut microbiota, growth, and survivability of three different treatment groups of Pacific white shrimp, Litopaneous vennamei, cultured in a recirculating system. The experiment was conducted over a 35-day trial period. Shrimps were fed diets, 15 times a day using automated feeder, supplemented with GOS at 0%, 0.15%, and 0.30% by weight. Denaturing gradient gel electrophoresis analysis revealed that the enteric microbial community of shrimp fed the basal diet differed markedly (<80.0% similarity coefficient) from those fed FOS-supplemented diets. However, shrimp survival, weight gain, and immune responses among the treatment groups were good but not significantly different (P > 0.05), probably due to the limited length of the feeding trial.


Assuntos
Aquicultura , Microbioma Gastrointestinal/efeitos dos fármacos , Oligossacarídeos/farmacologia , Penaeidae/efeitos dos fármacos , Ração Animal , Animais , Suplementos Nutricionais , Microbioma Gastrointestinal/imunologia , Oligossacarídeos/administração & dosagem , Penaeidae/crescimento & desenvolvimento , Penaeidae/imunologia
8.
Ecotoxicol Environ Saf ; 188: 109893, 2020 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-31735370

RESUMO

Cellular and humoral responses were evaluated in Litopenaeus vannamei juveniles when exposed to malathion, endosulfan, and their mixture. Each experiment was performed in the hemolymph collected at each exposure time (5 and 96 h) in duplicate; total hemocyte count, coagulation time, hemocyanin concentration, phenoloxidase (PO) and superoxide dismutase (SOD) activities were quantified. Survival was not affected by pesticides applied individually and mixed. Clotting time did not show significant differences concerning increase of concentration percentage of the pesticides tested. In organisms exposed to the pesticide mixture, hemocyanin decreased at 5 h of exposure as the concentration increased. Only in the malathion experiment did exposed shrimp to 10 and 50% of the LC50-96 h show significantly (p < 0.05) higher hemocyte contents. For malathion, significantly (p < 0.05) lower PO values in shrimp exposed to higher concentrations (10 and 50% of the LC50-96 h) were determined. While for the mixture treatment, high SOD value was determined at high exposure time and concentration. Malathion was the pesticide that showed an effect on some variables even at sublethal concentrations. The Continuous Concentration Criteria of the United States Environmental Protection Agency did not represent effects on the variables when they were compared with the averages of the control group.


Assuntos
Endossulfano/toxicidade , Imunidade Celular/efeitos dos fármacos , Imunidade Humoral/efeitos dos fármacos , Malation/toxicidade , Penaeidae/efeitos dos fármacos , Animais , Sinergismo Farmacológico , Hemocianinas/sangue , Hemócitos/efeitos dos fármacos , Hemócitos/imunologia , Hemolinfa/efeitos dos fármacos , Hemolinfa/imunologia , Monofenol Mono-Oxigenase/metabolismo , Penaeidae/imunologia
9.
Fish Shellfish Immunol ; 97: 294-299, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31863905

RESUMO

Crustins are cysteine-rich antimicrobial peptides (AMPs) widely distributed across crustaceans. From the four described crustin Types (I to IV), crustins from the subtype IIa are the most abundant and diverse members found in penaeid shrimp. Despite the critical role of Type IIa crustins in shrimp antimicrobial defenses, there is still limited information about their synthesis and antimicrobial properties. Here, we report the subcellular localization and the antibacterial spectrum of crusFpau, a Type IIa crustin from the pink shrimp Farfantepenaeus paulensis. The recombinantly expressed crusFpau showed antimicrobial activity against both Gram-positive and Gram-negative bacteria at low concentrations. Results from immunofluorescence using anti-rcrusFpau antiserum revealed that crusFpau is synthetized and stored by both granular and semigranular hemocytes, but not by hyaline cells. Interestingly, not all granular and semigranular hemocytes stained for crusFpau, revealing that this crustin is produced by specific granule-containing hemocyte subpopulations. Finally, we showed that the granule-stored peptides are not constitutively secreted into the plasma of healthy animals.


Assuntos
Antibacterianos/farmacologia , Peptídeos Catiônicos Antimicrobianos/biossíntese , Proteínas de Artrópodes/biossíntese , Hemócitos/metabolismo , Penaeidae/imunologia , Animais , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Penaeidae/metabolismo , Penaeidae/microbiologia
10.
Fish Shellfish Immunol ; 95: 498-505, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31698068

RESUMO

The study was to develop Vibrio harveyi biofilm-based novel microbial product and its oral delivery for high health Penaeus vannamei farming. Yield of bacterial biofilm was optimized on chitin substrate (size: <360, 360-850 and 850-1250 µm; concentration: 0.3, 0.6 and 0.9%) in tryptone soy broth (0.15%). The biofilm was characterized by crystal violet assay, SEM and LSCM imaging; protein profiling by SDS-PAGE and LC-ESI-MS/MS. The immune stimulatory effect of the biofilm in yard experiments was evaluated by relative quantification of immune genes using real-time PCR effect on overall improvement on health status under field trials. The highest biofilm yield (6.13 ±â€¯0.2 × 107 cfu/ml) was obtained at 0.6% of <360 µm chitin substrate. The biofilm formation was stabilized by 96 h of incubation at 30 °C. Protein profiling confirmed expression of six additional proteins (SDS-PAGE) and 11 proteins were differentially expressed (LC-ESI-MS/MS) in biofilm cells over free cells of V. harveyi. Oral administration of the biofilm for 48 h confirmed to enhance expression of antimicrobial peptides, penaeidin, crustin and lysozyme in P. vannamei. Further Oral administration of biofilm for two weeks to P. vannamei (1.8 ±â€¯0.13 g) improved the growth (2.66 ±â€¯0.06 g) and survival (84.44 ±â€¯1.82%) compared to control (2.15 ±â€¯0.03 g; 70.94 ±â€¯0.66%) Nursery trials showed a significant reduction in occurrence of anatomical deformities like antenna cut (12.67 ±â€¯0.66%), rostrum cut (4.66 ±â€¯0.87%), and tail rot (3.33 ±â€¯0.88%), compared to animals fed with normal diet which was 24.33 ±â€¯2.72; 14 ±â€¯1.52 and 10.66 ±â€¯1.45% respectively. In vitro and in vivo studies suggest inactivated biofilm cells of V. harveyi on chitin substrate express additional antigenic proteins and when administered orally through feed at regular intervals stimulates immune response and improve growth, survival and health status of shrimp.


Assuntos
Adjuvantes Imunológicos/administração & dosagem , Aquicultura , Biofilmes/crescimento & desenvolvimento , Penaeidae/imunologia , Penaeidae/microbiologia , Vibrio/imunologia , Administração Oral , Animais , Quitina/metabolismo , Alimentos Marinhos
11.
Fish Shellfish Immunol ; 95: 449-455, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31678535

RESUMO

Yellow head disease (YHD) is an infectious disease of Penaeus monodon which is caused by the yellow head virus (YHV). YHV infection invariably leads to 100% shrimp mortality within 3-5 days. Currently, an effective method to prevent or cure shrimp from YHV infection has not been elucidated. Therefore, the molecular mechanism underlying YHV infection should be examined. In this study, early endosome antigen 1 (EEA1) protein that was involved in the tethering step of the vesicle and early endosome fusion was investigated during YHV infection. The open reading frame of P. monodon EEA1 (PmEEA1) was cloned and sequenced (3000 bp). It encoded a putative protein of 999 amino acids and contained the zinc finger C2H2 domain signature at the N-terminus and the FYVE domain at the C-terminus. Suppression of PmEEA1 by specific dsRNA in shrimp showed inhibition of YHV replication after 48 h post YHV injection (hpi). On the other hand, shrimp received only NaCl without any dsRNA showed high YHV levels at approximately one hundred thousand times at 24 hpi and 48 hpi. Moreover, silencing of PmEEA1 by specific dsRNA followed by YHV challenge demonstrated a delay in shrimp mortality from 60 hpi to 168 hpi when compared to the control. These results indicated that YHV required PmEEA1 for trafficking within the infected cells, strongly suggesting that PmEEA1 may be a potential target to control and prevent YHV infection in P. monodon.


Assuntos
Interações Hospedeiro-Patógeno , Penaeidae/virologia , Roniviridae/patogenicidade , Proteínas de Transporte Vesicular/imunologia , Viroses/veterinária , Animais , Penaeidae/imunologia , Proteínas de Transporte Vesicular/genética , Viroses/imunologia , Replicação Viral
12.
Fish Shellfish Immunol ; 95: 670-678, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31689553

RESUMO

Palytoxins (PLTXs) are a group of complex and poisonous marine natural products that are toxic to marine life and even human beings. In the present study, the oxidative stress and immune response in the hepatopancreas and gills of Litopenaeus vannamei were assessed for 72 h after injection with PLTX extracts. Chemical and physiological parameters, e.g., the respiratory burst (O2-), activities of antioxidant enzymes, oxidative damage to lipids, carbonylation of proteins, and immune gene mRNA expression levels, were analysed. The results showed that the PLTX extract was not fatal to the shrimp but could reduce their mobility. The O2- levels in the gills gradually increased after exposure to PLTX extracts and were significantly higher than those in the control from 6 to 72 h. The malondialdehyde content, lipid peroxidation, protein carbonyl levels, and total antioxidant capacity in the gills all peaked at 12 h. At the same time, the gills were loosely connected, there was a clear disintegration of the epithelial tissue, and the stratum corneum disappeared after 12 h. In addition, compared to those in the control group, the PLTX extract treatment increased the O2- content, malondialdehyde content, lipid peroxidation, and protein carbonyl levels from 12 to 72 h, 24-48 h, 12-24 h, and 12-72 h after injection in the hepatopancreas of the shrimp, respectively. Both the Crustin and Toll gene expression levels significantly increased in the hepatopancreas compared to those in the control 6-72 h after injection of the toxin. In parallel, the expression levels of the manganese superoxide dismutase gene gradually decreased from 6 to 48 h and returned to normal levels after 72 h. Interestingly, the total antioxidant capacity also significantly increased compared to that in the control from 6 to 72 h. Our results indicate that although PLTX extracts cause lipid peroxidation and carbonylation of proteins in hepatopancreatic cells, leading to their damage, they did not cause a decrease in the total antioxidant capacity of the hepatopancreas.


Assuntos
Acrilamidas/administração & dosagem , Venenos de Cnidários/administração & dosagem , Dinoflagelados/química , Estresse Oxidativo , Penaeidae/efeitos dos fármacos , Penaeidae/imunologia , Acrilamidas/química , Animais , Venenos de Cnidários/química , Brânquias/efeitos dos fármacos , Hepatopâncreas/efeitos dos fármacos , Movimento , Oxirredução
13.
Food Funct ; 10(11): 7042-7051, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31580362

RESUMO

Tropomyosin is the most potent allergen of shrimp that can cause severe food allergy. However, to date, an effective approach to eliminate this allergenicity is still lacking. Glycation is a promising approach that can reduce the allergenicity of shrimp tropomyosin by destroying the epitopes; however, advanced glycation end products (AGEs) are also generated during glycation, which can function as neoallergens to strengthen the allergenicity; therefore, it is hard to tell how the glycation of an allergen with different saccharides affects the allergenicity via epitope loss and neoallergen generation. This study was aimed at the elucidation of how the glycation of tropomyosin (TM) with saccharides of different molecular sizes (glucose, maltose, and maltotriose) affected the allergenicity through epitope loss and the generation of neoallergns that belonged to advanced glycation end products (AGEs). Saccharides of higher molecular sizes (maltotriose) could lead to higher glycated TM than saccharides of smaller molecular sizes (glucose and maltose). Compared with TM, the TM glycated by glucose (TM-G) and maltotriose (TM-MTS) had lower allergenicity and contributed to weaker mouse allergy symptoms; on the other hand, the TM glycated by maltose (TM-M) had no significant impact on the allergenicity due to the generation of AGE-related neoallergens, which might offset the glycation-induced epitope loss. The glycation of TM by maltotriose led to lower generation of AGE neoallergens (e.g. CML) than that in the cases of glucose and maltose; therefore, maltotriose could be applied to desensitize TM-induced food allergy through glycation, and this could be a potential immunotherapy for shrimp allergy patients.


Assuntos
Produtos Finais de Glicação Avançada/imunologia , Penaeidae/imunologia , Hipersensibilidade a Frutos do Mar/imunologia , Tropomiosina/imunologia , Sequência de Aminoácidos , Animais , Epitopos/química , Epitopos/genética , Epitopos/imunologia , Feminino , Glucose/química , Produtos Finais de Glicação Avançada/química , Produtos Finais de Glicação Avançada/genética , Glicosilação , Humanos , Imunoglobulina E/imunologia , Maltose/química , Camundongos , Camundongos Endogâmicos BALB C , Penaeidae/química , Penaeidae/genética , Trissacarídeos/química , Tropomiosina/química , Tropomiosina/genética
14.
Fish Shellfish Immunol ; 94: 907-915, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31604147

RESUMO

Previous studies have demonstrated that white spot syndrome virus (WSSV) could induce hemocytes apoptosis in shrimps, however the inter-relationship between apoptotic process and the WSSV infection status is still currently underexplored. In the present work, the apoptosis and the viral proliferation in hemocytes of Litopenaeus vannamei were simultaneously investigated post WSSV infection by two-color immunofluorescence flow cytometry and real-time quantitative PCR. The apoptotic hemocytes of WSSV-infected shrimp was significantly increased at 12 h post infection (hpi), whereas underwent a slight decline at 24 hpi subsequently. Since 24 hpi, the apoptotic rate of hemocytes in the WSSV-infected shrimp exhibited a rapid and significant increase, and reached the peak level at 48 hpi with the ratio of 18.1 ±â€¯2.0%. Meanwhile, the percentage of WSSV-infected hemocytes and WSSV copies in hemocytes significantly increased at 24 hpi and maintained at a high level afterwards. With the rapid increase of hemocytes apoptosis, hemocyte density in hemolymph decreased dramatically to less than 20% of the mean value of control. Co-localization assay showed that the apoptotic WSSV-infected hemocytes occupied the dominant proportion of total apoptotic hemocytes, which reached the peak at 48 hpi with 12.6 ±â€¯1.5%. The expression profiles of seven pro-apoptotic genes and two apoptosis-inhibiting genes showed significant differential responses at different stages of WSSV infection, reflecting the interplay between the virus and the host immune response. Our results demonstrated that the apoptotic response of shrimp hemocytes could be significantly influenced by the WSSV infection process, which might provide an insight into deeper relationships between viral infection and apoptosis.


Assuntos
Apoptose , Proteínas de Artrópodes/genética , Hemócitos/imunologia , Penaeidae/imunologia , Vírus da Síndrome da Mancha Branca 1/fisiologia , Animais , Proteínas de Artrópodes/imunologia
15.
Fish Shellfish Immunol ; 95: 195-202, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31604149

RESUMO

Surface display can expose foreign antigenic protein on the surface of the vaccine vector, which is promising choice to elicit better immune responses. In this study, we apply this strategy to develop an immunoactivator by using a live attenuated Vibrio harveyi as an antigenic protein carrier with surface displayed VP28, a major envelope protein of white spot syndrome virus (WSSV), for two major pathogens of Litopenaeus vannamei. As a result, the immunoactivator showed self-limited growth and attenuation of virulence in shrimp via different inoculation routes either with single-repetitive dose or high dose. Moreover, either intramuscular injection or oral administration of the immunoactivator did not affect growth of shrimp body weight or cause pathologic changes. Additionally, the rapid immunoprotection was induced by the immunoactivator after administration for one week with highly relative percent survival (RPS) more than 90% against both V. harveyi and WSSV. Until 4 weeks post administration, the immunoactivator still possessed efficient immune effect with no less than 60% RPS for both pathogens. Totally, the attenuated V. harveyi surface displaying VP28 could be a potential immunoactivator for WSSV and vibriosis control in L. vannamei.


Assuntos
Penaeidae/imunologia , Vibrioses/veterinária , Vibrio/imunologia , Proteínas do Envelope Viral/imunologia , Viroses/veterinária , Vírus da Síndrome da Mancha Branca 1/imunologia , Animais , Vacinas Bacterianas/imunologia , Vibrio/genética , Vibrioses/prevenção & controle , Proteínas do Envelope Viral/genética , Vacinas Virais/imunologia , Viroses/prevenção & controle , Vírus da Síndrome da Mancha Branca 1/genética
16.
Fish Shellfish Immunol ; 95: 16-24, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31585243

RESUMO

The intestine is important for nutrition, metabolism and immunity. Succinate (SA) plays a vital role in the physiological homeostasis of animal intestines. However, the effects of dietary SA on the intestinal immunity and metabolism in shrimp are not clear. In this study, we investigated the immune and metabolic responses in the intestine of Litopenaeus vannamei that were fed diets consisting of different levels of SA: 0 g/kg (Con) and 10 g/kg (SA) for 56 days. The results from a RNA-seq analysis identified 6005 differentially expressed genes (DEGs), including 2728 upregulated genes and 3277 downregulated genes, which were grouped into 312 pathways. The DEGs were most enriched in pathways related to protein synthesis and amino acid metabolism, including "ribosome", "aminoacyl-tRNA biosynthesis", "pyrimidine metabolism", and "arginine and proline metabolism"; additionally, carbohydrate and lipid metabolism pathways were also activated. A large number of immune-related genes were associated with mucus barrier modification, antimicrobial activity, pathogen attachment and recognition, antioxidant activity, and apoptosis. The expression patterns of several candidate genes involved in the immune response and nutrition metabolism were detected by qPCR. This study provides insight into the transcriptomic modulating mechanisms associated with intestinal immunity and the metabolism of L. vannamei in response to the intake of dietary SA.


Assuntos
Imunidade Inata , Intestinos/imunologia , Penaeidae/imunologia , Penaeidae/metabolismo , Ácido Succínico/administração & dosagem , Transcriptoma , Ração Animal , Animais , RNA-Seq
17.
Carbohydr Polym ; 226: 115254, 2019 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-31582076

RESUMO

Three trials were conducted to comprehensively evaluate the application of chitooligosaccharides (COSs) different dosages, molecular weights (MW) and degrees of deacetylation (DD) as a feed additive for Pacific white shrimp (Litopenaeus vannamei). In trial 1, COSs (3 KDa and 85% DD) at four different dosages (0.05%, 0.1%, 0.15% and 0.2%) were added to feed to investigate the appropriate dosages. The survival ratio (SR) of the shrimps was not significantly different (P > 0.05) between the control and treatment groups. The shrimps fed with 0.1% COSs supplementation exhibited the highest wet body weight (FBW), specific growth ratio (SGR), and weight gain (WG) and the lowest feed conversion ratio (FCR). In trial 2, COSs with different MW (85% DD and 0.1% dosage) were tested. Except for the group treated with the 12 KDa COSs, all shrimps fed with COSs had remarkably higher (P < 0.05) FBW, WG, and SGR and lower FCR (P < 0.05) than the control group, and shrimps fed with the 1 KDa COSs showed most positive effects. In trial 3, COSs with different DD (MW of 1 KDa and 0.1% dosage) were further studied. The different DD were DD5, DD25, DD50, DD75, and DD95. Shrimps fed COSs-supplemented diets of DD75 or DD95 exhibited higher (P < 0.05) FBW, WG and SGR and lower FCR than the other groups. The DD95 group had the highest FBW, WG and SGR and the lowest FCR, but there were no significant differences (P > 0.05) between the DD75 and DD95 groups. Moreover, for COSs supplementation, especially for the DD75 and DD95 groups, the antioxidant parameters were significantly different from those of the other groups. Furthermore, groups fed diets with COSs supplementation had higher (P < 0.05) trypsin activity than those fed control diets. In addition, immune and antioxidant gene expression and the morphology of the hepatopancreas were affected by the DD of COSs. Therefore, the additive dosages, molecular weights and degrees of deacetylation of COSs significantly affected the growth performance of the shrimps; therefore, it is particularly important to determine the optimum parameters of COSs.


Assuntos
Quitina/análogos & derivados , Hepatopâncreas/metabolismo , Penaeidae , Ração Animal , Animais , Quitina/química , Quitina/metabolismo , Peso Molecular , Penaeidae/crescimento & desenvolvimento , Penaeidae/imunologia , Penaeidae/metabolismo
19.
Ecotoxicol Environ Saf ; 184: 109626, 2019 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-31536848

RESUMO

Of late, Pacific white shrimp Penaeus vannamei culture has intensified globally and is a major contributor to the cultured shrimp produced worldwide. Intensification of its culture has led to elevated ammonia concentration during grow-out. Ammonia toxicity is a function of water pH, temperature, salinity and beyond the optimum range, creates stress to cultured aquatic species which can reduce growth, increase susceptibility to diseases and eventually mortality. The present study was aimed at quantifying the toxic effect of total ammonia nitrogen (TAN) (1, 3, 6 & 9 mg/l) and pH levels (6, 8 & 10) individually and in combination on median survival (50% lethal time) of shrimp (8 g) after exposure for 14 days followed by post-stress challenge with white spot syndrome virus (WSSV) for 9 days. Mortality risk factor and the toxicity effect on the immune variables were evaluated. Individual stressors showed a risk factor of 1-13 times, whereas combined treatments considerably increased the risk of dying compared to control. Low survival (15%) was observed in pH6TAN9 and pH10TAN3 treatments and was substantiated by prominent histological obliteration in gills of shrimp. The cumulative mortality in post-stress WSSV challenged trials was 1-5 times and 1-35 times in individual and combination treatments, respectively compared to control. The study revealed that variations in ammonia and pH beyond the optimal range significantly influence the non-specific immune mechanisms in P.vannamei and increases the susceptibility to WSSV especially in combination treatments.


Assuntos
Amônia/toxicidade , Penaeidae/efeitos dos fármacos , Penaeidae/imunologia , Estresse Salino , Poluentes Químicos da Água/toxicidade , Vírus da Síndrome da Mancha Branca 1/fisiologia , Animais , Concentração de Íons de Hidrogênio , Compostos de Nitrogênio/toxicidade , Penaeidae/virologia , Estresse Salino/imunologia , Análise de Sobrevida
20.
Fish Shellfish Immunol ; 94: 497-509, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31541775

RESUMO

As a crucial neuroendocrine-immune factor, dopamine (DA) could regulate the immune system of Litopenaeus vannamei. To understand the immune mechanisms and regulatory pathways of DA in L. vannamei, the transcriptome analysis of hemocytes of L. vannamei with injection of DA (10-6 mol/shrimp) at 3 and 12 h were performed in this study. Moreover, quantitative real-time PCR (qPCR) method was applied to validate the accuracy of transcriptome sequencing and analyze the expression pattern of candidate differentially expressed genes (DEGs) at different time points (0, 3, 6, 12, and 24 h) after DA injection. The results showed that a total of 51382 unigenes with a N50 length of 2341 bp were generated. And 1397 and 457 DEGs were obtained by comparative transcriptome at 3 and 12h respectively. Moreover, the results of functional annotation and enriched pathway showed that the DEGs were involved in phagosome (ko04145), lysosome (ko04142), Endocytosis (ko04144), and NOD-like receptor signaling pathway (ko04621). Besides, the Pearson's correlation coefficient (R) between transcriptome sequencing and qPCR was 0.845, which confirmed the reliability of the transcriptome sequencing results and the accuracy of assembly. Furthermore, the expression pattern of 15 candidate DEGs, containing 9 up-regulated and 6 down-regulated DEGs at 3 h, indicated the regulation of DA in physiological functions especially in the immune system. Therefore, these results revealed that DA induced the expressions of membrane receptors or proteins, activated intracellular signaling pathways, regulated cellular and humoral immune systems, controlled antioxidation and apoptosis, and was involved in the regulation of neuroendocrine system. These findings are helpful to promote the understanding on the effects of biogenic amines on physiological functions and regulatory networks of crustacean, and offer a substantial material and foundation for researching the immune response of crustacean.


Assuntos
Dopaminérgicos/metabolismo , Dopamina/metabolismo , Hemócitos/imunologia , Imunidade Inata/genética , Penaeidae/imunologia , Transcriptoma/efeitos dos fármacos , Animais , Dopamina/administração & dosagem , Dopaminérgicos/administração & dosagem , Perfilação da Expressão Gênica , Imunidade Inata/efeitos dos fármacos , Penaeidae/genética
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