Changes in the anxiety-like and fearful behavior of shrimp following daily threatening experiences.

Behavioral variation in animals is often influenced by experience. Previous studies have found that daily threatening experiences can enhance fear- and anxiety-like behaviors in some vertebrates. However, it is unclear whether the change in fear/anxiety behavior occurs in invertebrates. The present study investigated whether fear/anxiety behavior could be affected by a net-chasing treatment in two shrimp species (Neocaridina denticulata ssp. and Palaemon pacificus). The net-chasing treatment was repeated for 8 days to simulate daily predator experiences, and behavioral tests (open-field, shelter-seeking, and escape-response tests) were conducted on the day following the last day of treatment. Net-chased N. denticulata ssp. displayed a tendency to remain near a wall compared with the control in the open-field test, whereas net-chased P. pacificus shrimps demonstrated greater escape behavior compared with the control in the escape-response test. These results suggest that fear/anxiety behavior for both shrimp species can be affected by the net-chasing treatment, although the pattern of behavioral change differed between the two species. The findings suggest that daily threatening experiences change the behavior of shrimp and cause them to select a regular avoidance strategy when they encounter risks and unknown situations.

Transcriptome analysis of Neocaridina denticulate sinensis challenged by Vibrio parahemolyticus.

As a common aquatic pathogen, Vibrio parahaemolyticus can cause a variety of diseases of shrimp, especially acute hepatopancreatic necrosis disease (AHPND), which leads to great losses to the aquaculture industry around the world. However, the molecular mechanism of V. parahaemolyticus infection is still unclear. Neocaridina denticulate sinensis is a kind of small ornamental shrimp that is popular in aquarium trade, and due to its tenacious vitality, rapid growth, high reproductive capacity, it is very suitable to be developed as an animal model for basic research on decapod crustaceans. Thus, in this paper, transcriptomes of N. denticulate sinensis hepatopancreas with or without V. parahaemolyticus injection were explored. The results showed that a total of 23,624 genes with the N50 of 2705 bp were obtained. Comparative transcriptomic analysis revealed 21,464 differentially expressed genes between the V. parahaemolyticus infected and non-infected group, of which, 11,127 genes were up-regulated and 10,337 genes were down-regulated. Functional enrichment analysis suggested that many DEGs enriched in immune related pathways, including MAPK signaling pathway, Phosphatidylinositol signaling system, Chemokine signaling pathway, Phagosome and Jak-STAT signaling pathway and so on. Eight genes were selected randomly for qRT-PCR to verify the transcriptome sequencing results and the results showed the expression of these genes were consistent with the transcriptome results. Our work provides a unique and important dataset that contributes to the understanding of the molecular mechanisms of the immune response to V. parahaemolyticus infection and may further provide the basis for the prevention and resolution of bacterial diseases.

Transcriptomic analysis of Neocaridina denticulate sinensis hepatopancreas indicates immune changes after copper exposure.

Neocaridina denticulate sinensis is a promising crustacean model species due to its merits in raising and breeding. However, its molecular responses to copper remains largely unknown. In the present research, RNA-seq was used to mine the alteration in transcriptome of N. denticulate sinensis hepatopancreas under copper exposure. A total of 16,423 DEGs was identified between control and Cu2+ treatment groups. GO enrichment analysis of all DEGs suggested down-regulated genes exceeded up-regulated genes in all the significantly enriched terms, except for RNA polymerase III complex (GO:0005666). KEGG analysis showed Cu exposure only induced two significantly enriched pathways, including Phagosome (ko04145) and Pathogenic Escherichia coli infection (ko05130). Besides, pattern recognition receptors as Toll, lectin B, CTL1 and SRB, AMPs as crustin type I, lysozyme, and NOS were down-regulated after Cu2+ exposure, while hemocyanin, MT, HSP70 and HSP90 were significantly up-regulated, implying these molecules may play vital role in Cu2+ detoxification of N. denticulate sinensis. Our results here provide research direction of heavy metal detoxification of N. denticulate sinensis, simultaneously enriched its genomic information.

De novo assembly transcriptome analysis reveals the genes associated with body color formation in the freshwater ornamental shrimps Neocaridina denticulate sinensis.

The body color of Neocaridina denticulate sinensis is a compelling phenotypic trait, in which a cascade of carotenoid metabolic processes plays an important role. The study was conducted to compare the transcriptome of cephalothoraxes among three pigmentation phenotypes (red, blue, and chocolate) of N. denticulate sinensis. The purpose of this study was to explore the candidate genes associated with different colors of N. denticulate sinensis. Nine cDNA libraries in three groups were constructed from the cephalothoraxes of shrimps. After assembly, 75022 unigenes were obtained in total with an average length of 1026 bp and N50 length of 1876 bp. There were 45977, 25284, 23605, 21913 unigenes annotated in the Nr, Swissprot, KOG, and KEGG databases, respectively. Differential expression analysis revealed that there were 829, 554, and 3194 differentially expressed genes (DEGs) in RD vs BL, RD vs CH, and BL vs CH, respectively. These DEGs may play roles in the absorption, transport, and metabolism of carotenoids. We also emphasized that electron transfer across the inner mitochondrial membrane (IMM) was a key process in pigment metabolism. In addition, a total of 6328 simple sequence repeats (SSRs) were also detected in N. denticulate sinensis. The results laid a solid foundation for further research on the molecular mechanism of integument pigmentation in the crustacean and contributed to developing more attractive aquatic animals.

Searching and identifying pigmentation genes from Neocaridina denticulate sinensis via comparison of transcriptome in different color strains.

Aquaria species are characterized by their amazing colors and patterns. Research on the breeding molecular genetics of ornamental shrimps is surprisingly limited. We conducted a transcriptome analysis to investigate the expression of encoding genes in the integument of the strains Neocaridina denticulate sinensis. After assembled and filtered, 19,992 unigenes were annotated by aligning with public functional databases (NR, Swiss-Prot, KEGG, COG). 14,915 unigenes with significantly different expressions were found by comparing three strains integument transcriptomes. Ribosomal protein genes, ABC transporter families, calmodulin, carotenoid proteins and crustacyanin may play roles in the cytological process of pigment migration and chromatophore maintenance. Numerous color genes associated with multiple pathways including melanin, ommochrome and pteridines pathways were identified. The expression patterns of 25 candidate genes were analysis by qPCR in red, yellow, transparent and glass strains. The qPCR results in red, yellow and transparent were consistent with the level of RPKM values in the transcriptomes. The above results will advance our knowledge of integument color varieties in N. denticulate sinensis and help the genetic selection of crustaceans with consumer-favored colors. Furthermore, it also provides some candidate pigmentation genes to investigate the correlation between coloration and sympatric speciation in crustaceans.

Transcriptome analysis of Neocaridina denticulate sinensis under copper exposure.

Neocaridina denticulate sinensis is a small freshwater economic shrimp, as well as excellent laboratory model for their short life cycle and easy availability. However, the response of N. denticulate sinensis to pervasive copper pollution in aquatic environments has not been deeply investigated yet. Herein, we preformed Illumina sequencing technology to mine the alterations of cephalothorax transcriptome under 2.5 µmol/L of Cu2+ after 48 h. 122,512 unigenes were assembled and 219 unigenes were identified as significantly differentially expressed genes (DEGs) between control and Cu2+ treatment groups. Functional enrichment analysis revealed that DEGs were mostly associated with immune responses and molting, such as endocytosis, Fc gamma R-mediated phagocytosis and chitin metabolic process. Seven genes were chosen for qPCR verification, and the results showed that the transcriptome sequencing data were consistent with the qPCR results. This is the first report of transcriptome information about N. denticulate sinensis. These results provided a direction for the future research of resistance to Cu2+ in this shrimp, and simultaneously enriched gene information of N. denticulate sinensis.

Ecosystem metabolism regulates seasonal bioaccumulation of metals in atyid shrimp (Neocaridina denticulata) in a tropical brackish wetland.

Natural dissolved organic matter (DOM) forms the base of aquatic food webs and is a key environmental factor that affects the bioavailability of metals for aquatic organisms. Aquatic communities are naturally exposed simultaneously to environments containing a mixture of metals and varying DOM levels and compositions. However, the exact effect of DOM on metal bioaccumulation is difficult to predict due to temporal and spatial variations in sources, production, and consumption of DOM, and to interactions between DOM and metals. Ecosystem metabolism describes the process of organic carbon production and consumption and, therefore, the trophic status of ecosystems. However, whether and how ecosystem metabolism determines the seasonality of metal bioaccumulation remains unclear. The present study used in-situ water quality sondes and discrete field samplings to establish the relationship between the seasonality of ecosystem metabolism; related environmental and limnological regulators; the metal speciation and concentration in bulk water and sediments; and their metal bioaccumulation. The target population consisted of atyid shrimp (Neocaridina denticulata) in a brackish constructed wetland in tropical Taiwan was sampled between August 2014 and November 2015. Metal bioaccumulation displayed distinct seasonal patterns that peaked in summer (Cu, Cd, Cr, Zn, Mn, and Se) or winter (Pb and Ni). The in situ production (gross primary production) and heterotrophic consumption (ecosystem respiration) of organic matter significantly decreased with increasing waterborne DOM levels in this heterotrophic wetland. Both dissolved free metals bioavailable for respiratory surfaces (As, Zn, Cu, and Cr) and insoluble metals available for dietary intake (Mn and Ni) decreased with increasing DOM, as well as with decreasing gross primary production and ecosystem respiration. Seasonal variations of metal bioaccumulation also paralleled the transition in wetland trophic status, which reflected the effect of potential qualitative changes in the wetland DOM pool. Bioaccumulation of most metals displayed strong correlations with gross primary production, ecosystem respiration, and wetland trophic status. Our findings demonstrated that ecosystem metabolism can play a key mediating role in the seasonality of metal bioaccumulation in atyid shrimp, as it links the variation and interaction between DOM level/source, the speciation/bioavailability, and the uptake efficiency for metals by aquatic organisms. This study contributes to the temporal-specific risk assessment of aquatic metal exposure in regional environmental settings. It also reveals ecosystem-specific spectra in the context of changes in climate and environment.

Acute toxicity of mixture of acetaminophen and ibuprofen to Green Neon Shrimp, Neocaridina denticulate.

In recent years, numerous studies have indicated that various long-term use drugs, such as antibiotics or analgesics, not only cannot be completely decomposed via sewage treatment but also exhibit biological toxicity if they enter the environment; thus, the release of these drugs into the environment can damage ecological systems. This study sought to investigate the acute toxicity of two commonly utilized analgesics, ibuprofen (IBU) and acetaminophen (APAP), to aquatic organisms after these drugs have entered the water. To address this objective, the acute toxicity (median lethal concentration, LC50, for a 96-h exposure) of IBU alone, APAP alone, and mixtures containing different ratios of IBU and APAP in green neon shrimp (Neocaridina denticulata) were measured. The results of four tests revealed that the 96-h LC50 values for IBU and APAP alone were 6.07 mg/L and 6.60 mg/L, respectively. The 96-h LC50 for a 1:1 mixture of IBU and APAP was 6.23 mg/L, and the toxicity of this mixture did not significantly differ from the toxicity of either drug alone (p<0.05). The experimental results for mixtures containing unequal ratios of IBU and APAP indicated that mixtures with high APAP concentrations and low IBU concentrations exhibited markedly greater toxicity in N. denticulata (LC50=4.78 mg/L) than APAP or IBU alone. However, mixtures with high IBU concentrations and low APAP concentrations exhibited lower toxicity in N. denticulata (LC50=6.78 mg/L) than IBU or APAP alone. This study demonstrated that different mixtures of IBU and APAP were associated with different toxic effects in green neon shrimp.