Effects of ciprofloxacin on Eichhornia crassipes phytoremediation performance and physiology under hydroponic conditions.

Antibiotics can be absorbed by aquatic plants, but they seriously affect the health of aquatic plants and threaten the steady state of aquatic ecosystem. The phytoremediation performance and physiology of floating macrophyte (Eichhornia crassipes) under antibiotic ciprofloxacin (CIP) hydroponic conditions were investigated. It was found that CIP absorption of E. crassipes was up to 84.38% and the root was the main absorption tissue. Hydrolysis and microbial degradation were the second removal pathway of CIP followed the plant absorption. After 7 days of CIP exposure, the photosynthesis efficiency of E. crassipes remained stable, and the presence of CIP did not inhibit the growth of the plant. On the 14th day, the superoxide dismutase and catalase activities were increased in response to the CIP stress. However, the tender leaves of E. crassipes turned white and shrivel, attributed to a decrease in chlorophyll content and chlorophyll fluorescence parameters after 21 days of CIP exposure. These findings will have significant implications for E. crassipes to absorb CIP on a limited time-scale and provide a phytoremediation technology for antibiotics in water.

Molecular characterization of carboxypeptidase B-like (CPB) in Scylla paramamosain and its role in white spot syndrome virus and Vibrio alginolyticus infection.

Carboxypeptidase plays an important physiological role in the tissues and organs of animals. In this study, we cloned an entire 2316 bp carboxypeptidase B-like (CPB) sequence with a 1302 bp open reading frame encoding a 434 amino acid peptide from Scylla paramamosain. The CPB gene was expressed highly in hepatopancreas and decreased in crab hemocytes after challenges with white spot syndrome virus (WSSV) or Vibrio alginolyticus. After CPB gene knockdown using double-stranded RNA (CPB-dsRNA), the expression of JAK, STAT, C-type lectin, crustin antimicrobial peptide, Toll-like receptors, prophenoloxidase, and myosin II essential light chain-like protein were down-regulated in hemocytes at 24 h post dsRNA treatment. CPB knockdown decreases total hemocyte count in crabs indicated that CPB may negatively regulate crab hemocyte proliferation in crabs. CPB showed an inhibitory effect on hemocyte apoptosis in crabs infected with WSSV or V. alginolyticus. The phagocytosis rate of WSSV by hemocytes was increased after CPB-dsRNA treatment. After WSSV challenge, the mortality and WSSV copy number were both decreased but the rate of hemocyte apoptosis was increased in CPB-dsRNA-treated crabs. The results indicate that the antiviral activity of the crabs was enhanced when CPB was knocked down, indicating WSSV may take advantage of CPB to benefit its replication. In contrast, the absence of CPB in crabs increased mortality following the V. alginolyticus challenge. The phagocytosis rate of V. alginolyticus by hemocytes was increased after CPB-dsRNA treatment. It was revealed that CPB may play a positive role in the immune response to V. alginolyticus through increasing the phagocytosis rate of V. alginolyticus. This research further adds to our understanding of the CPB and identifies its potential role in the innate immunity of crabs.

Hesperetin protects crayfish Procambarus clarkii against white spot syndrome virus infection.

Hesperetin is a natural flavanone compound, which mainly exists in lemons and oranges, and has potential antiviral and anticancer activities. In this study, hesperetin was used in a crayfish pathogen challenge to discover its effects on the innate immune system of invertebrates. The crayfish Procambarus clarkii was used as an experimental model and challenged with white spot syndrome virus (WSSV). Pathogen challenge experiments showed that hesperetin treatment significantly reduced the mortality caused by WSSV infection, while the VP28 copies of WSSV were also reduced. Quantitative reverse transcriptase polymerase chain reaction revealed that hesperetin increased the expression of several innate immune-related genes, including NF-kappaB and C-type lectin. Further analysis showed that hesperetin treatment plays a positive effects on three immune parameters like total hemocyte count, phenoloxidase and superoxide dismutase activity. Nevertheless, whether or not infected with WSSV, hesperetin treatment would significantly increase the hemocyte apoptosis rates in crayfish. These results indicated that hesperetin could regulate the innate immunity of crayfish, and delaying and reducing the mortality after WSSV challenge. Therefore, the present study provided novel insights into the potential therapeutic or preventive functions associated with hesperetin to regulate crayfish immunity and protect crayfish against WSSV infection, provide certain theoretical basis for production practice.

Molecular characterization of minichromosome maintenance protein (MCM7) in Scylla paramamosain and its role in white spot syndrome virus and Vibrio alginolyticus infection.

The minichromosome maintenance protein (MCM7) is a member of the MCM protein family which participates in the MCM complex by playing a role in the cell replication cycle and chromosome initiation in eukaryotes. The 2270 bp cDNA sequence of MCM7, including a 2127-bp open reading frame (ORF) encoding a 709-aa protein, was cloned from Scylla paramamosain using RT-PCR and RACE. Data showed that MCM7 was highly expressed in the digestive organ and hepatopancreas of S. paramamosain. Furthermore, MCM7 expression was down-regulated by infection with white spot syndrome virus (WSSV) or Vibrio alginolyticus. When MCM7 was knocked down, immune genes such as Janus kinase (JAK) and crustin antimicrobial peptide (CAP) were down-regulated, and C-type-lectin (CTL) was up-regulated in hemocytes. The mortality of WSSV-infected or V. alginolyticus-infected crabs was enhanced following MCM7 knockdown. It was demonstrated that MCM7 is very important in the progression of WSSV and V. alginolyticus infection. We also investigated the effect of MCM7 on apoptosis rate and phagocytic rate in S. paramamosain. MCM7 knockdown caused higher levels of apoptosis in the hemocytes of the control, WSSV, and V. alginolyticus groups. MCM7 knockdown influenced the activity of phenoloxidase (PO) and superoxide dismutase (SOD), and total hemocyte count (THC) after infection with WSSV or V. alginolyticus, which indicated that MCM7 plays a regulatory role in innate immunity of crabs. Thus, we conclude that MCM7 may participate in the anti-WSSV and V. alginolyticus immune response in crabs by regulating apoptosis and the activity of PO and SOD.

Comparative transcriptomic analysis of crab hemocytes in response to white spot syndrome virus or Vibrio alginolyticus infection.

To assess the immune response of hemocytes to white spot syndrome virus (WSSV) or Vibrio alginolyticus infection in the mud crab Scylla paramamosain, a transcriptome analysis was performed. We report the analysis of 45131 transcripts from S. paramamosain hemocytes by de novo assembly. A comparison with GenBank protein and nucleotide sequences identified 33699 genes as previously known. The length distribution of the genes was 8147 genes ≥200 bp, 4714 genes ≥300 bp, and 3517 genes ≥2000 bp. A total of 21579 simple sequence repeats (SSRs) were found in the transcriptomic dataset, including 9% monomers, 53.34% dimers and 32.55% trimers. A total of 13172 and 5087 differentially expressed transcripts were found in the V. alginolyticus-infected group and WSSV-infected group, respectively. Of these, 5920 transcripts were up-regulated and 7252 were down-regulated in the V. alginolyticus-infected crabs and 2302 transcripts were up-regulated and 2785 were down-regulated in the WSSV-infected crabs. Additionally, 3096 transcripts were differentially expressed simultaneously in the V. alginolyticus-infected crabs and the WSSV-infected crabs. Several known immune-related genes such as heat shock protein, Janus kinase, STAT, relish, caspase, Ca2+-transporting ATPase and lysosomal alpha-mannosidase were found among the differentially expressed transcripts. Transcription and its regulation were significant biological processes, and ATP binding and zinc ion binding were significant molecular functions. This is the first report of comparative transcriptomic analysis of crab hemocytes in response to WSSV or V. alginolyticus infection. These findings will contribute to our understanding of the immune response to WSSV and V. alginolyticus infection in crustaceans.

Molecular characterization of shrimp harbinger transposase derived 1 (HARBI1)-like and its role in white spot syndrome virus and Vibrio alginolyticus infection.

The role of the nuclease, HARBI1-like protein (mjHARBI1-like) in the innate immunity of Marsupenaeus japonicus was explored in this study. The 1361 bp cDNA sequence of mjHARBI1-like was cloned from M. japonicus using RACE. RT-qPCR analysis results showed that the gills and hepatopancreas of M. japonicus were the main tissues where mjHARBI1-like is expressed. In addition, it was also found that white spot syndrome virus (WSSV) or Vibrio alginolyticus challenge could stimulate mjHARBI1-like expression. After mjHARBI1-likewas inhibited, expression of immune genes such as toll, p53, myosin, and proPO were significantly downregulated (P < 0.01). However, in shrimp hemocytes, hemocyanin and tumor necrosis factor-α (TNF-α) were up-regulated significantly (P < 0.01). This study demonstrated that mjHARBI1-like plays a key role in the progression of WSSV and V. alginolyticus infection. Specifically, the cumulative mortality of WSSV-infected and V. alginolyticus-infected shrimp was significantly advanced by double-strand RNA interference (dsRNAi) of mjHARBI1-like. Apoptosis studies indicated that mjHARBI1-dsRNA treatment caused a reduction in hemocyte apoptosis in bacterial and viral groups. In addition, phagocytosis experiments illustrated that mjHARBI1-dsRNA treatment led to a lower phagocytosis rate in hemocytes of V. alginolyticus-challenged shrimp. It was also found that knockdown of mjHARBI1-like inhibited shrimp phenoloxidase (PO) activity, superoxide dismutase (SOD) activity, and total hemocyte count (THC) after WSSV or V. alginolyticus infection. These data indicate a regulative role of mjHARBI1-likein the immunity of shrimp in response to pathogen infection. Resultantly, it was concluded that mjHARBI1-like might have a positive effect on the anti-WSSV immune response of shrimp by regulating apoptosis, THC, PO activity, and SOD activity. Additionally, mjHARBI1-like might promote anti-V. alginolyticus infection by participating in regulating phagocytosis, apoptosis, SOD activity, PO activity, and THC.