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1.
Gene ; 788: 145583, 2021 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-33753150

RESUMO

Macrobrachium nipponense has the characteristics of fast ovarian development cycle, which leads to the coexistence of multiple generations, the reduction of commodity specifications and the low economic benefit. Therefore, the study on the mechanism of ovarian development is of great significance to the development of industry. Cyclin A (CycA)is a key gene regulating ovarian development in vertebrates, but little information was available for its function in crustaceans. In this study, the full-length cDNA of Mn-CycA was obtained from the ovary. The full-length cDNA (2033 bp) with an open reading frame of 1368 bp, encoded a 456-amino acid protein. qRT-PCR revealed tissue-specific expression pattern of Mn-CycA, with abundant expression in the ovary. Results in different developmental stages of ovary indicated that Mn-CycA expression is positively correlated with ovarian maturation. qRT-PCR In different developmental stages, the expression of Mn-CycA mRNA gradually increased during the embryonic stage and decreased significantly on the first day of the hatching stage. At the 25th day of the metamorphosis stage, the expression level of Mn-CycAmRNA in female shrimp was 3.5 times higher than that in male shrimp, which may be related to the proliferation of oogonia and the formation of oocytes. In situ hybridization (ISH) of ovary showed Mn-CycA was examined in all stages and was mainly located in oogonia and oocytes. Compared with the control group, the obvious change of gonad somatic index (GSI) proved that injection of Mn-CycA dsRNA could delay the ovarian development cycle, which provided strong evidence for the involvement of Mn-CycA in ovarian maturation and oogenesis, and expanded a new perspective for studying the fast ovarian development cycle in M. nipponense.


Assuntos
Ciclina A/genética , Ciclina A/metabolismo , Perfilação da Expressão Gênica/métodos , Palaemonidae/crescimento & desenvolvimento , Animais , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/metabolismo , Sequência de Bases , Clonagem Molecular , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Masculino , Oócitos/crescimento & desenvolvimento , Oócitos/metabolismo , Oogônios/crescimento & desenvolvimento , Oogônios/metabolismo , Fases de Leitura Aberta , Especificidade de Órgãos , Palaemonidae/genética , Palaemonidae/metabolismo , Filogenia
2.
Ecotoxicol Environ Saf ; 208: 111767, 2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-33396085

RESUMO

Ammonia nitrogen and nitrite are two common forms of environmental toxicants for aquatic organisms including crustaceans. The PI3K-AKT pathway is an important intracellular signaling pathway related to cellular stress response, but involvement of this pathway in the immunotoxicological response of decapod crustaceans to aquatic toxicants such as ammonia nitrogen and nitrite still remains enigmatic. In this study, based on transcriptome mining and molecular cloning techniques, three key genes (named as MrPI3K, MrAKT and MrFoxO) in the PI3K-AKT signaling pathway were identified from the giant river prawn Macrobrachium rosenbergii. Sequence homology and phylogenetic analysis revealed that all the three genes harbored signature sequences of corresponding protein families, and shared high levels of similarities with their respective homologs from other species. MrPI3K, MrAKT and MrFoxO all displayed ubiquitous tissue distribution profiles, but their expression levels varied to a great extend among different tissues and between sexes. Following exposure to nitrite (20 mg/L nitrite-N) or ammonia (25 mg/L total ammonia-N) stresses for 24 h and 48 h, the three genes all responded by altering their expression levels at different time points, but they didn't show uniform expression patterns following these stresses, indicating the diversified roles of these genes in different tissues and the complexity of this signaling pathway. Remarkably, MrPI3K and MrAKT were induced only in the hemocytes and intestine, respectively, indicating their specific roles in these organs. Our study demonstrated the potential utility of these genes as biomarkers of acute ammonia or nitrite toxicity in prawns, and also provided evidence that the PI3K-AKT pathway is involved in the immunotoxicological responses to nitrite and ammonia stress in M. rosenbergii.


Assuntos
Amônia/toxicidade , Nitritos/toxicidade , Palaemonidae/fisiologia , Proteínas Proto-Oncogênicas c-akt/genética , Poluentes Químicos da Água/toxicidade , Animais , Decápodes/metabolismo , Hemócitos/metabolismo , Nitrogênio/metabolismo , Palaemonidae/metabolismo , Penaeidae/metabolismo , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/metabolismo , Filogenia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais , Transcriptoma
3.
Ecotoxicol Environ Saf ; 208: 111591, 2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-33396114

RESUMO

The Palaemonid shrimp Palaemon macrodactylus is widely distributed in coastal areas and estuaries which are easily contaminated by various pollutants. However, the responses of this species to environmental toxicants are not well described. In the present study, adult individuals of P. macrodactylus were exposed to gradient concentrations of Cadmium (Cd) to evaluate its acute toxic effects, including bioaccumulation, induced oxidative stress and changed energy metabolism in this species. The medium lethal concentration (LC50) of Cd at 24 h, 48 h, 72 h, and 96 h were 2.60, 0.88, 0.49 and 0.37 mg/L, respectively. Cd bioaccumulations in tissues of shrimp increased in a concentration-dependent manner, and higher concentration (50% 96 h-LC50, 0.185 mg/L) of Cd exposure led to a maximum increase of Cd concentration by 14.8, 145.5 and 15.8 folds in gill, hepatopancreas and abdominal muscle. Cd exposure caused a significant inhibition on the activity of catalase (CAT), and total superoxide dismutase (T-SOD), decrease in the total antioxidant capacity (T-AOC), and an increase of malonadehyde (MDA) content, which indicated a damage to the antioxidant system of shrimp. Meanwhile, Cd exposure also led to a significant up-regulation in the expression level of metallothionein gene (MT), and down-regulations at the mRNA level of heat shock protein 70 (HSP70) and CAT. Moreover, Cd exposure significantly inhibited the oxygen consumption rate (22%), and increased the ammonia excretion rate (43%), hence lead to a significant decrease of the O:N ratio (45%) in shrimp. The results indicated that Cd exposure could induce obvious oxidative stress, energy metabolic dysfunction and bioaccumulation of Cd in P. macrodactylus. The data obtained from the present study would provide useful information for further understanding on the toxicological mechanism of Cd to crustaceans in coastal areas and estuaries.


Assuntos
Cádmio/toxicidade , Palaemonidae/fisiologia , Poluentes Químicos da Água/toxicidade , Animais , Antioxidantes/metabolismo , Bioacumulação , Catalase/metabolismo , Estuários , Brânquias/metabolismo , Proteínas de Choque Térmico HSP70/metabolismo , Hepatopâncreas/metabolismo , Metalotioneína/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Palaemonidae/metabolismo , Superóxido Dismutase/metabolismo
4.
PLoS One ; 15(12): e0243778, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33362263

RESUMO

The giant freshwater prawn, Macrobrachium rosenbergii (M. rosenbergii) as an important freshwater aquaculture species with high commercial value, exhibited unsynchronized growth. However, the potentially metabolic mechanism remains unclear. In this study, we used liquid chromatography tandem mass spectrometry (LC-MS/MS) to investigate the hepatopancreatic metabolic profiles of twenty giant freshwater prawns between the fast-growing group and slow-growing group. In the metabolomics assay, we isolated 8,293 peaks in positive and negative iron mode. Subsequently, 44 significantly differential metabolites were identified. Functional pathway analysis revealed that these metabolites were significantly enriched in three key metabolic pathways. Further integrated analysis indicated that glycerophospholipid metabolism and aminoacyl-tRNA biosynthesis have significant impact on growth performance in M.rosenbergii. Our findings presented here demonstrated the critical metabolites and metabolic pathways involved in growth performance, moreover provided strong evidence for elucidating the potentially metabolic mechanism of the unsynchronized growth in M. rosenbergii.


Assuntos
Ductos Biliares , Metabolômica , Palaemonidae/crescimento & desenvolvimento , Palaemonidae/metabolismo , Pâncreas , Animais
5.
PLoS One ; 15(8): e0237037, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32810189

RESUMO

The genus Palaemon comprises worldwide marine and freshwater shrimps and prawns, and some of them are ecologically or commercially important species. Palaemon is not currently a monophyletic group, so phylogenetics and systematics are constantly changing. Species crypticism has been pointed out in several Palaemon species, being the clearest evidence in the European rockpool shrimp P. elegans. Here we sequenced and described seven European Palaemon mitochondrial genomes. The mitochondrial protein-coding genes were used, along with those of three other Palaemon species, to perform mitogenome phylogenetic analyses to clarify the evolutionary relationships within the genus, and particularly to shed light on the cryptic species found within P. elegans. The Messinian Salinity Crisis (5.3-5.9 Ma, late Miocene) was proposed to be the origin of this cryptic species and it was used as aged constraint for calibration analysis. We provide the largest and the first time-calibrated mitogenome phylogeny of the genus Palaemon and mitogenome substitution rate was estimated (1.59% per million years) in Decapoda for the first time. Our results highlighted the need for future systematics changes in Palaemon and crypticism in P. elegans was confirmed. Mitochondrial genome and cox1 (1.41%) substitution rate estimates matched those published elsewhere, arguing that the Messinian Salinity Crisis was a plausible event driving the split between P. elegans and its cryptic species. Molecular dating suggested that Pleistocene glaciations were likely involved in the differentiation between the Atlantic and Mediterranean populations of P. elegans. On the contrary, the divergence between the Atlantic and Mediterranean populations of the common littoral shrimp P. serratus was greater and dated to be much older (4.5-12.3 Ma, Plio-Miocene), so we considered that they could represent two separated species. Therefore, species crypticism in the genus Palaemon seems to be a common phenomenon.


Assuntos
Genoma Mitocondrial/genética , Palaemonidae/genética , Animais , Sequência de Bases/genética , Evolução Biológica , DNA Mitocondrial/genética , Decápodes/genética , Palaemonidae/metabolismo , Filogenia
6.
Mol Immunol ; 126: 1-7, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32712503

RESUMO

Rel/nuclear factor (NF)-κB family of transcription factors paly vital roles in innate immunity response to bacterial and viral infection. Here, we cloned and identified a dorsal homologue (named as MnDorsal) from Macrobrachium nipponense. The full-length cDNA of MnDorsal is 2573 bp with a 1986 bp open reading frame that encodes 661 amino acids. Predicted MnDorsal protein contained a RHD (Rel homology domain), an IPT (Iglike, plexins, and transcriptions factors) domain, and two low complexity regions. Phylogenetic analysis showed that MnDorsal has a closer genetic distance with dorsal homologues from invertebrates. MnDorsal was widely expressed in a variety of tissues, including hemocytes, heart, hepatopancreas, gills, stomach, and intestine. Expression patterns analysis showed that the transcriptional level of MnDorsal in the gills was evidently up-regulated after Staphylococcus aureus, Vibrio parahaemolyticus, white spot syndrome virus, or polyinosinic-polycytidylic acid challenge, suggesting that MnDorsal participates in the immune defenses against pathogens and stimulant challenges. Additionally, the dsRNA-mediated RNA interference analysis showed that knockdown of MnDorsal can significantly inhibit the expression of anti-lipopolysaccharide factor (ALF) and crustin. Further studies revealed that the up-regulated expression of ALFs (MnALF2, MnALF3, and MnALF4) and crustins (MnCrustin3 and MnCrustin4) caused by S. aureus infection were obviously decreased after silencing MnDorsal. These findings suggest that MnDorsal positively regulate the expression of antibacterial peptides (AMPs) during S. aureus infection. Our study will promote to better understand the role of Toll-Dorsal-AMPs pathway in innate immunity response to gram-positive bacterial infection in crustacean.


Assuntos
Peptídeos Catiônicos Antimicrobianos/genética , Proteínas de Artrópodes/metabolismo , Imunidade Inata/genética , Palaemonidae/imunologia , Fatores de Transcrição/metabolismo , Animais , Peptídeos Catiônicos Antimicrobianos/imunologia , Aquicultura , Proteínas de Artrópodes/genética , Regulação da Expressão Gênica/imunologia , Técnicas de Silenciamento de Genes , Palaemonidae/genética , Palaemonidae/metabolismo , Palaemonidae/microbiologia , Poli I-C/imunologia , Staphylococcus aureus/imunologia , Fatores de Transcrição/genética , Vibrio parahaemolyticus/imunologia , Vírus da Síndrome da Mancha Branca 1/imunologia
7.
Gene ; 754: 144891, 2020 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-32535048

RESUMO

Characterized by ankyrin repeat motifs, the feminization-1 (fem-1) gene plays an essential role in sex determination/differentiation in Caenorhabditis elegans. However, there are only a few reports on fem-1 in crustaceans. In this study, a fem-1 gene (Mrfem-1) was first isolated from the giant freshwater prawn Macrobrachium rosenbergii. The full-length cDNA of Mrfem-1 was 2607 bp long, containing an open reading frame encoding 615 amino acids, and presenting eight ankyrin repeats. The full-length cDNA has been submitted to GenBank with the accession no. MT160093. According to the RT-PCR results, Mrfem-1 was exclusively expressed in the ovary. The expression level of Mrfem-1 had increased with ovarian maturation and reached the highest peak at vitellogenic stage. In situ hybridization results showed that positive signals were concentrated in the cytoplasm of previtellogenic stage, and scattered in the cytoplasm and follicular cells at vitellogenic stage, suggesting that Mrfem-1 might be associated with ovarian maturation. Moreover, two effective siRNAs targeting Mrfem-1 were found and their effectiveness verified in vitro. These results on Mrfem-1 will help us to better understand the fem family and provide a new resource for subsequent investigations of siRNA-mediated regulation on ovarian development in M. rosenbergii.


Assuntos
Proteínas de Ciclo Celular/metabolismo , Regulação da Expressão Gênica , Ovário/crescimento & desenvolvimento , Ovário/metabolismo , Palaemonidae/metabolismo , RNA Interferente Pequeno/genética , Animais , Sequência de Bases , Proteínas de Ciclo Celular/antagonistas & inibidores , Proteínas de Ciclo Celular/genética , Feminino , Especificidade de Órgãos , Palaemonidae/genética , Filogenia
8.
Gen Comp Endocrinol ; 293: 113478, 2020 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-32243957

RESUMO

This study identified an insulin-like peptide (ILP) in Macrobrachium rosenbergii termed Mr-ILP and further investigated its function through glucose injection and RNAi. With the analysis of five other glucose metabolism related genes, this study shed light on the molecular mechanism of carbohydrate metabolism in crustaceans. Mr-ILP shared the typical skeleton with six conserved cysteine and mainly expressed in neuroendocrine system. In M. rosenbergii, the elevated hemolymph glucose concentration after glucose injection returned to basal levels in short time, implying an efficient regulatory system in carbohydrate metabolism. Hyperglycemic related genes answered the elevated hemolymph glucose concentration quickly with significant decreased expression level, while Mr-ILP showed delayed response. Instead, glycolysis increased after glucose injection, which indicated glycolysis might play an important role in lowering the abnormally high glucose level. In vivo silencing of Mr-ILP, by injecting the prawns with double-stranded RNA (dsRNA) for 21 days reduced its expression by approximately 75%. Accordingly, glycogen synthase decreased and the trehalose and glycogen level in the hepatopancreas were significantly reduced, indicating the function of Mr-ILP in oligosaccharide and polysaccharide accumulation. When Mr-ILP was silenced, the expression of hyperglycemic related genes were enhanced, but the hemolymph glucose level was not elevated significantly, which might attribute to the increased glycolysis to keep a balanced glucose level in hemolymph.


Assuntos
Metabolismo dos Carboidratos , Insulina/metabolismo , Palaemonidae/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Metabolismo dos Carboidratos/genética , DNA Complementar/genética , Feminino , Regulação da Expressão Gênica , Glucose/administração & dosagem , Hemolinfa/metabolismo , Insulina/química , Insulina/genética , Masculino , Palaemonidae/genética , Filogenia
9.
PLoS One ; 15(2): e0229171, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32084182

RESUMO

Glutathione peroxidase (GPx) has been the focus of increased research because of its important role as an antioxidant and in reactive oxygen species (ROS) induced damage repair. Studies on GPxs have relevance with Macrobrachium nipponense because it has poor tolerance to hypoxia in Macrobrachium nipponense. The two subunits named as MnGPx-3 and MnGPx-4 according to the glutathione peroxidase nomenclature system. Both full-length cDNAs were cloned from the hepatopancreas. In this study, we analyzed the expression of two GPxs in Macrobrachium nipponense in response to changes in environmental oxygen. Expression levels of MnGPx-3 and MnGPx-4 indicated that both have strong responses to hypoxia. In situ hybridization showed that MnGPx-3 and MnGPx-4 were located in secretory and storage cells in hepatopancreas. These results suggest that GPx gene is expressed and released by secretory cells and released response to hypoxia. In the gill tissue, however, GPxs are located in blood cells, suggesting that they perform different functions in different tissues or organs. The results of in situ hybridization were consistent with those of quantitative Real-time PCR. This study provides a basis for understanding the oxidative stress response in M. nipponense under hypoxia.


Assuntos
Regulação Enzimológica da Expressão Gênica , Glutationa Peroxidase/genética , Oxigênio/metabolismo , Palaemonidae/genética , Palaemonidae/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , Glutationa Peroxidase/química , Hibridização In Situ , Palaemonidae/enzimologia , Filogenia , RNA Mensageiro/genética , Distribuição Tecidual
10.
Fish Shellfish Immunol ; 98: 414-419, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31962148

RESUMO

Anti-lipopolysaccharide factors (ALFs), as main host-defense molecules of crustaceans, form a unique family of antimicrobial peptides (AMPs). MnALF4 is one isoform of ALFs isolated from the freshwater prawn Macrobrachium nipponense. In the present study, MnALF4 gene was successfully expressed in the yeast Pichia pastoris and the recombinant MnALF4 protein exhibited efficient and broad-spectrum antimicrobial activities against both Gram-positive bacteria and Gram-negative bacteria in vitro. When prawns were injected with rMnALF4 before bacterial challenge with E. coli, the recombinant protein effectively promote the elimination of bacteria by the host. It manifested that rMnALF4 could effectively kill the invading bacteria in vivo. Treatment with rMnALF4 led to remarkable changes in bacterial morphology, such as spheroidization, oversized bacteria, and cell lysis. In addition, rMnALF4 showed weak hemolysis activity to the rabbit red blood cells. Our work suggests that MnALF4 plays an important role in Macrobrachium immunity and is worthy of further investigation as a potential antibacterial agent with high efficacy against bacterial infection and low toxicity to host cells.


Assuntos
Proteínas de Artrópodes/metabolismo , Lipopolissacarídeos/imunologia , Palaemonidae/metabolismo , Pichia/metabolismo , Proteínas Recombinantes/metabolismo , Animais , Anti-Infecciosos/metabolismo , Anti-Infecciosos/farmacologia , Proteínas de Artrópodes/química , Proteínas de Artrópodes/genética , Bactérias/efeitos dos fármacos , Eritrócitos/efeitos dos fármacos , Regulação da Expressão Gênica , Hemólise/efeitos dos fármacos , Pichia/genética , Coelhos , Proteínas Recombinantes/genética
11.
Bull Environ Contam Toxicol ; 104(2): 222-227, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31893301

RESUMO

This paper assesses the role of intestinal helminth parasite of Macrobrachium vollenhovenii on host metal accumulation in Lekki lagoon, Lagos, Nigeria. Lead, zinc, copper, chromium and iron concentrations were determined in water, sediment, intestinal parasite, intestine of infected and non-infected prawns. Intestinal nematode parasite Cucullanus sp. was identified. Pb and Cr were not detected in the water and sediment. The sediment had higher concentrations of the metals than the water medium, and exceeded the WHO limits for Zn, Cu and Fe. Cucullanus sp. accumulated Zn 19 times, Cr several thousand times more and Fe 11 times more than the host intestine (wet wt). This parasite could have accumulated these metals from its host, reducing the concentration of these metals in the tissues of the infected prawns compared to non-infected ones. The infected and non-infected prawns accumulated Zn and Fe several times more than the water medium. The parasite influenced metal accumulation in the host tissue and pose ecological risk.


Assuntos
Metais Pesados/metabolismo , Nematoides/fisiologia , Palaemonidae/parasitologia , Poluentes Químicos da Água/metabolismo , Animais , Monitoramento Ambiental , Sedimentos Geológicos/química , Mucosa Intestinal/metabolismo , Intestinos/parasitologia , Metais Pesados/análise , Nigéria , Palaemonidae/metabolismo , Água/química , Poluentes Químicos da Água/análise
12.
Arch Environ Contam Toxicol ; 78(2): 245-253, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31858197

RESUMO

Open-air landfill's may be are considered as a potential source of human environmental exposure to chemical substances such as, polycyclic aromatic hydrocarbons and toxic metals. Due to possible availability of mercury in the environment caused by open landfill emissions, this study evaluates the spatiality and seasonality of macroinvertebrates, in particular shrimps (Macrobrachium amazonicum), exposure to mercury (Hg). Information regarding Hg accumulation in this crustacean may be important for the development of public policies aiming conservation and preservation of ecosystems surrounding landfills in Amazon, and around the world. Sampling occurred quarterly in the following months: November/2015; February/2016; May/2016 and; August/2016. In each of these months, three points were selected: P1, P2 and P3. The samples were processed via acid digestion and the quantification of metal was performed by inductively coupled plasma mass spectrometry. The average concentration of total mercury (T-Hg) was 24.565 ± 6.610 µg kg-1 wet weight, with minimum and maximum limits of 12.742 ± 11.367 (P3) and 35.509 ± 14.761 µg kg-1 wet weight (P1) in November/2015 and August/2016, respectively. The concentration of total mercury (T-Hg) in shrimps was different between points (p = 0.004) and months (p = 0.000). The T-Hg concentrations were significantly higher in May and August 2016, which corresponds to the dry season. The presence of landfills promotes large accumulation of T-Hg in the aquatic biota and represents a risk to human health. However, seasonal changes in T-Hg levels were observed. In the wettest period, bioconcentration factor levels decrease in aquatic organisms.


Assuntos
Monitoramento Ambiental/métodos , Mercúrio/análise , Palaemonidae/efeitos dos fármacos , Poluentes Químicos da Água/análise , Animais , Organismos Aquáticos/efeitos dos fármacos , Organismos Aquáticos/metabolismo , Bioacumulação , Brasil , Ecossistema , Biomarcadores Ambientais , Mercúrio/farmacocinética , Mercúrio/toxicidade , Músculos/efeitos dos fármacos , Músculos/metabolismo , Palaemonidae/metabolismo , Rios , Estações do Ano , Instalações de Eliminação de Resíduos , Poluentes Químicos da Água/farmacocinética , Poluentes Químicos da Água/toxicidade
13.
Artigo em Inglês | MEDLINE | ID: mdl-31846692

RESUMO

The oriental river prawn, Macrobrachium nipponense, is a commercial freshwater prawn species in China. It is highly sensitive to hypoxia, and this has posed a challenge to its intensive culturing. To date, the effects of hypoxia on reproduction in female prawns are not entirely clear, as are the underlying mechanisms of the effects of hypoxia. In this work, comparative transcriptome and gene expression analyses of the eyestalk were performed in M. nipponense females under hypoxia and reoxygenation conditions. Sequencing and de novo assembly of the combined reads yielded 43,583 unigenes with an average length of 1726 bp. A total of 711 genes were found to be differentially expressed in the eyestalk under the hypoxia and reoxygenation conditions. With the help of functional and pathway enrichment analysis of the differentially expressed genes, a novel set of transcripts that were associated with several important functions, such as hormone biosynthesis and progesterone-mediated oocyte maturation, were identified. Additionally, ten neuropeptides were identified based on the differentially expressed transcripts, and they were validated by quantitative real-time polymerase chain reaction (qRT-PCR) and reverse transcription PCR (RT-PCR) analyses. Three neuropeptide genes were expressed in the neural tissue and ovary of the prawns; this indicates that they were involved in reproductive processes. In particular, RNA interference (RNAi) short neuropeptide F dramatically promoted ovary maturation, as indicated by the gonad somatic index. While the present findings do indicate that hypoxia affects reproductive function in M. nipponense females, in-depth functional analyses of the candidate neuropeptides should be conducted in the future to understand their role in hypoxia adaptation and the associated mechanisms that affect the reproductive capacity of this species.


Assuntos
Palaemonidae/genética , Palaemonidae/metabolismo , Animais , Proteínas de Artrópodes/metabolismo , Olho/metabolismo , Feminino , Hipóxia/metabolismo , Neuropeptídeos/metabolismo , Oxigênio/administração & dosagem , Oxigênio/metabolismo , Palaemonidae/efeitos dos fármacos , Transcriptoma
14.
Curr Microbiol ; 77(1): 40-48, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31659402

RESUMO

Some patients with candidiasis seek alternatives drug to treat vaginal yeast infection like herbal preparations and probiotics. However, the effectiveness of such treatments has not received much study. In this research, the unique chitinotrophic Bacillus was isolated on shrimp shell from salt lakes and identified as Bacillus altitudinis by 16SRNA sequencing. This strain produced a novel chitin-oligosaccharide material and thermostable chitinase (5.1 units/ml) during 4 days incubation on shrimp shell medium; nevertheless, its growth on nutrient agar was negative. The zymogram showed less than 50 kD protein responsible for chitinase activities. The LC/MS detection of concentrate fermented products showed the production of oligosaccharide during chitin fermentation. As results of shrimp shell degradation, 65.6 mg/l protein, 73.4 mg/l N-acetyl glucose amine, and oligosaccharide were produced. Synergism activities of chitooligosaccharide and chitinase from this strain against fungi and pathogen candida (staining with methylene blue showed that almost 50% of 106 cells were died during 6 h) are promising for new anti-fungal drug with no side effect.


Assuntos
Antifúngicos/farmacologia , Bacillus/metabolismo , Quitinases/farmacologia , Oligossacarídeos/farmacologia , Exoesqueleto/metabolismo , Animais , Antifúngicos/metabolismo , Candida albicans/efeitos dos fármacos , Candida glabrata/efeitos dos fármacos , Cromatografia Líquida , Feminino , Fermentação , Humanos , Espectrometria de Massas , Palaemonidae/metabolismo
15.
Bull Environ Contam Toxicol ; 103(5): 729-733, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31531704

RESUMO

Accumulation of nonylphenol (NP) in hepatopancreas, gonad, eyestalk, and muscle of freshwater prawn Macrobrachium rosenbergii following 72 h exposure to 100 µg/L NP, and depuration of NP in these tissues at 0.5-192 h post exposure were examined. We also examined the expressions of vitellogenin (Vg) and vitellogenin receptor (VgR) of prawn following 0-20 days exposure to 0, 1, 10, and 100 µg/L NP. NP accumulation in hepatopancreas and gonad with high concentration, and low concentration in muscle, but depurated faster in eyestalk and muscle. The expressions of vitellogenin (Vg) and vitellogenin receptor (VgR) increased directly with dose and time. In conclusion, NP accumulated significantly in gonad together with high Vg and VgR expressions, and depurated slow in hepatopancreas and gonad when prawns were removed back to control water. The induction of Vg and VgR under NP exposure might be a stress response in M. rosenbergii.


Assuntos
Proteínas do Ovo/genética , Água Doce/química , Palaemonidae/efeitos dos fármacos , Fenóis/toxicidade , Receptores de Superfície Celular/genética , Vitelogeninas/genética , Poluentes Químicos da Água/toxicidade , Animais , Bioacumulação/efeitos dos fármacos , Relação Dose-Resposta a Droga , Expressão Gênica/efeitos dos fármacos , Gônadas/efeitos dos fármacos , Gônadas/metabolismo , Hepatopâncreas/efeitos dos fármacos , Hepatopâncreas/metabolismo , Taxa de Depuração Metabólica , Palaemonidae/metabolismo , Fenóis/metabolismo , Poluentes Químicos da Água/metabolismo
16.
Bull Environ Contam Toxicol ; 103(5): 689-696, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31506728

RESUMO

To test the toxic effects of tributyltin (TBT), Macrobrachium rosenbergii were exposed to three concentrations of TBT viz. 10 ng/L, 100 ng/L and 1000 ng/L for 90 days. The bioaccumulation of TBT level varied in hepatopancreas based upon dose dependent manner. Histopathological results revealed the reduction in basement membrane thickness, disruption of the hepatopancreatic tubules and abnormal lumen in hepatopancreas of TBT treated prawns. The ultrastructure of the control prawn showed normal architecture of cellular organelles with prominent nuclei in hepatocytes. On the other hand, many vacuoles, irregular arrangements of microvilli, swollen mitochondria, distorted rough endoplasmic reticulum cisternaes and abnormal nucleus were seen in the TBT treated group. Further, the biochemical and vitellogenin content were altered remarkably due to TBT exposure. It directly indicated that TBT had conspicuously inhibited the vitellogenesis. Therefore, it was inferred that the administration of TBT has considerably affected the hepatopancreatic functions in M. rosenbergii.


Assuntos
Bioacumulação , Água Doce/química , Hepatopâncreas/efeitos dos fármacos , Palaemonidae/efeitos dos fármacos , Compostos de Trialquitina/toxicidade , Poluentes Químicos da Água/toxicidade , Animais , Relação Dose-Resposta a Droga , Hepatopâncreas/metabolismo , Palaemonidae/metabolismo , Palaemonidae/ultraestrutura , Compostos de Trialquitina/metabolismo , Vitelogeninas/metabolismo , Poluentes Químicos da Água/metabolismo
17.
Fish Shellfish Immunol ; 93: 223-231, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31319208

RESUMO

The oriental river prawn, Macrobrachium nipponense, is an economically and nutritionally important species of the Palaemonidae family of decapod crustaceans. Ammonia is a major aquatic environmental pollutant that negatively affects the health of prawns and their associated commercial productivity. Here, we used high-throughput sequencing techniques for detecting the effects of ammonia stress (22.1 mg/L ammonia-N for 48 h) on gene expression in the hepatopancreas of M. nipponense. We generated 176,228,782 high-quality reads after eliminating adapter sequences and filtering out low-quality reads, which were assembled into 63453 unigenes. Comparative analysis of the expression profiles of the ammonia-treated and control groups identified 887 differentially expressed genes (P < 0.05), including 481 upregulated genes and 406 downregulated genes. Analyses of the GO and KEGG databases revealed significant differences between the two groups in 32 pathways. Immune-related pathways under ammonia stress included Complement and coagulation cascades, Platelet activation, B cell receptor signaling pathway, Antigen processing and presentation, Chemokine signaling pathway, NOD-like receptor signaling pathway, RIG-I-like receptor signaling pathway, T cell receptor signaling pathway and Toll-like receptor signaling pathway. Remarkably, ammonia stress altered the expression patterns of key immune genes (lectin3, syntenin, alpha-2-macroglobulin, cathepsin L, PIM3, serine protease inhibitor, suppressor of cytokine signaling-2 like protein), indicating that ammonia-stress induce immune response. These data provide new insights into the immune response of M. nipponense and pave a new way for fighting ammonia stress. The genes and pathways identified here represent valuable genetic resources for development of molecular markers and genetic breeding studies.


Assuntos
Amônia/efeitos adversos , Proteínas de Artrópodes/genética , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Palaemonidae/efeitos dos fármacos , Transcriptoma/efeitos dos fármacos , Poluentes Químicos da Água/efeitos adversos , Animais , Proteínas de Artrópodes/metabolismo , Perfilação da Expressão Gênica , Hepatopâncreas/efeitos dos fármacos , Hepatopâncreas/metabolismo , Palaemonidae/genética , Palaemonidae/metabolismo , Transcriptoma/genética
18.
Artigo em Inglês | MEDLINE | ID: mdl-30936021

RESUMO

We examined the effects of exogenous dopamine on gill (Na+, K+)-ATPase activity in vitro in microsomal preparations from juvenile or adult freshwater shrimp, Macrobrachium amazonicum. Dopamine had no effect on enzyme activity in juveniles but stimulated activity in adult shrimp gills by ≈35%. Stimulation of the gill (Na+, K+)-ATPase in adult shrimps by 100 mmol L-1 dopamine was characterized kinetically by varying ATP, MgATP, and Na+ and K+ concentrations, together with inhibition by ouabain. Dopamine stimulated ATP hydrolysis by ≈40% obeying Michaelis-Menten kinetics, reaching VM = 190.5 ±â€¯15.7 nmol Pi min-1 mg-1 protein, KM remaining unaltered. Stimulation by Na+ (≈50%) and K+ (≈25%) revealed distinct kinetic profiles: although KM values were similar, Na+ stimulation followed cooperative kinetics, contrasting with the Michaelian kinetics seen for K+. Stimulation by MgATP increased activity by ≈30% with little change in KM. Similar saturation profiles were seen for ouabain inhibition with very similar calculated KI values. Our findings suggest that dopamine may be involved in hemolymph sodium homeostasis by directly binding to the gill (Na+, K+)-ATPase at a site different from ouabain, thus stimulating enzyme activity in an ontogenetic stage-specific manner. However, dopamine binding does not affect enzyme affinity for cations and ouabain. This is the first report of the direct action of dopamine in stimulating the crustacean gill (Na+, K+)-ATPase.


Assuntos
Dopamina/farmacologia , Brânquias/efeitos dos fármacos , ATPase Trocadora de Sódio-Potássio/metabolismo , Trifosfato de Adenosina/metabolismo , Animais , Água Doce , Brânquias/metabolismo , Palaemonidae/efeitos dos fármacos , Palaemonidae/metabolismo , Potássio/metabolismo , Sódio/metabolismo , ATPase Trocadora de Sódio-Potássio/química
19.
Int J Mol Sci ; 20(7)2019 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-30965605

RESUMO

The doublesex and mab-3 related transcription factor (DMRT) gene family involvement in sex development is widely conserved from invertebrates to humans. In this study, we identified a DM (Doublesex/Mab-3)-domain gene in Macrobrachium nipponense, which we named MniDMRT11E because it has many similarities to and phylogenetically close relationships with the arthropod DMRT11E. Amino acid alignments and structural prediction uncovered conservation and putative active sites of the DM domain. Real-time PCR analysis showed that the MniDMRT11E was highly expressed in the ovary and testis in both males and females. Cellular localization analysis showed that DMRT11E was mainly located in the oocytes of the ovary and the spermatocyte of the testis. During embryogenesis, the expression level of MniDMRT11E was higher at the cleavage stage than at other stages. During the different stages of ovarian development, MniDMRT11E expression gradually increased from OI to OIII and decreased to the lowest level at the end of OIV. The results indicated that MniDMRT11E probably played important roles in embryonic development and sex maturity in M. nipponense. MniDMRT11E dsRNA injection also significantly reduced vitellogenin (VG) expression and significantly increased insulin-like androgenic gland factor (IAG) expression, indicating a close relationship in gonad development.


Assuntos
Proteínas de Artrópodes/metabolismo , Palaemonidae/embriologia , Palaemonidae/metabolismo , Animais , Proteínas de Artrópodes/genética , Feminino , Regulação da Expressão Gênica no Desenvolvimento/genética , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Masculino , Ovário/embriologia , Ovário/metabolismo , Palaemonidae/genética , Testículo/embriologia , Testículo/metabolismo
20.
Fish Shellfish Immunol ; 90: 363-375, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30974219

RESUMO

Superoxide dismutases (SODs) are important antioxidant enzymes that occur in virtually all oxygen-respiring organisms, and copper/zinc SOD (Cu/ZnSOD) is one of the most important SODs. In the present study, Macrobrachium rosenbergii Cu/Zn-SOD was expressed in a yeast eukaryotic system. The open reading frame (ORF) of MrCu/ZnSOD was cloned into the plasmid vector pHAC181, and the recombinant plasmid was integrated into the downstream region of the GAL1 promoter in Saccharomyces cerevisiae strain GAL1-ScRCH1 via homologous recombination. The resulting recombinant MrCu/ZnSOD consisted of a 3 × HA-tag at its C-terminal. Via western blot, the molecular weight of the recombinant MrCu/ZnSOD was estimated at about 30 kDa. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values of this recombinant MrCu/ZnSOD ranged from 0.556 to 0.840 µM, and from 0.967 to 2.015 µM, respectively. The recombinant MrCu/ZnSOD protein was able to agglutinate four Gram-negative bacterial strains, as well as two of three Gram-positive strains (except Staphylococcus aureus). This demonstrated that the recombinant protein possessed some antimicrobial activity against certain Gram-positive and Gram-negative bacteria. M. rosenbergii were fed with the recombinant yeast strain MrCu/ZnSOD for 4 weeks and then challenged with the most common crustacean pathogen, Vibrio parahaemolyticus. This group of prawns presented lower mortality, higher enzymatic activity, and higher expression of the mRNA of immune-related genes than that in the control groups. Taken together, these results suggest that MrCu/ZnSOD is an antioxidant enzyme and antimicrobial peptide involved in the crustacean innate immune system and offers protection to the host against pathogenic bacteria.


Assuntos
Palaemonidae/genética , Palaemonidae/imunologia , Superóxido Dismutase-1/genética , Superóxido Dismutase-1/imunologia , Vibrio parahaemolyticus/imunologia , Aglutinação , Animais , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/imunologia , Proteínas de Artrópodes/metabolismo , Bactérias Gram-Negativas/imunologia , Bactérias Gram-Positivas/imunologia , Microrganismos Geneticamente Modificados/metabolismo , Palaemonidae/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Superóxido Dismutase-1/metabolismo
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