Gloria Yepiz-Plascencia

Shrimp thioredoxin is a potent antioxidant protein

Thioredoxin (TRX) is a main component of the redox homeostasis machinery in the cell and it is required for ribonucleotide reductase function among others. In invertebrates, the redox balance is compromised during disease and changes in the physiological state and it is one of the components of the innate immune response. In this work, the shrimp (Litopenaeus vannamei) LvTRX cDNA was sequenced, cloned and over-expressed in bacteria to further characterize the function of the recombinant protein. LvTRX was able to reduce insulin disulfides and it was a better antioxidant compared to reduced glutathione and ascorbic acid, by means of the Trolox Equivalent Antioxidant Capacity (TEAC) assay. Interestingly, LvTRX contains aside of the canonical active site CXXC disulfide motif, one Cys (C73) residue in the interface of a putative dimer previously reported for human TRX. Using qRT-PCR, we found that shrimp LvTRX is mainly expressed in gills and pleopods; the variation of LvTRX mRNA upon hypoxia and re-oxygenation is not statistically significant. LvTRX stands as an important antioxidant that must be considered in future physiological and immune challenges studies.

Expression and activity of glutathione S-transferases and catalase in the shrimp Litopenaeus vannamei inoculated with a toxic Microcystis aeruginosa strain

Microcystin (MC) produced during cyanobacteria blooms is notably toxic to human and wildlife. Conjugation with reduced glutathione (GSH) by glutathione S-transferase (GST) and the antioxidant enzymes defenses (e.g. catalase, CAT) are important biochemical defense mechanisms against MCs toxicity. We investigated the enzymatic activity of CAT and GST and the gene expression levels of CAT and eight GST isoforms in the hepatopancreas of the globally farmed shrimp Litopenaeus vannamei 48-h after injection with a sub-lethal dose of 100 μg kg⁻¹ of a toxic Microcystis aeruginosa extract. MCs caused up-regulation for GSTΩ, μ and a MAPEG isoform, by 12-, 2.8- and 1.8-fold, respectively, and increases in the total GST enzyme activity and CAT enzyme activity. The study points to the importance of further characterization for the L. vannamei GST isoforms and GST/CAT post-translational regulation processes to better understand the key mechanisms involved in the shrimps defense against MC exposure.

The complete mitochondrial genomes of the yellowleg shrimp Farfantepenaeus californiensis and the blue shrimp Litopenaeus stylirostris (Crustacea: Decapoda).

Mitochondria play key roles in many cellular processes. Description of penaeid shrimp genes, including mitochondrial genomes are fairly recent and some are focusing on commercially important shrimp as the Pacific shrimp Litopenaeus vannamei that is being used for aquaculture not only in America, but also in Asia. Much less is known about other Pacific shrimp such as the yellowleg shrimp Farfantepenaeus californiensis and the blue shrimp Litopenaeus stylirostris. We report the complete mitogenomes from these last two Pacific shrimp species. Long DNA fragments were obtained by PCR and then used to get internal fragments for sequencing. The complete F. californiensis and L. stylirostris mtDNAs are 15,975 and 15,988 bp long, containing the 37 common sequences and a control region of 990 and 999 bp, respectively. The gene order is identical to that of the tiger shrimp Penaeus monodon. Secondary structures for the 22 tRNAs are proposed and phylogenetic relationships for selected complete crustacean mitogenomes are included. Phylogenomic relationships among five shrimp show strong statistical support for the monophyly of the genus across the analysis. Litopenaeus species define a clade, with close relationship to Farfantepenaeus, and both clade with the sister group of Penaeus and Fenneropenaeus.

A novel functional glucose transporter in the white shrimp Litopenaeus vannamei -LvGLUT2- is up-regulated during hypoxia in hepatopancreas.

In hypoxia conditions, the white shrimp Litopenaeus vannamei shifts its energetic metabolism from aerobic to anaerobic, requiring more glucose uptake into the cells by GLUT proteins. We here report a novel glucose transporter in shrimp. The Lvglut2 cDNA is 2473 bp-long containing an ORF of 1458 bp encoding 486 amino acid residues. The deduced protein has the features of a facilitative sugar transporter. The Lvglut2 gene product tagged with GFP was expressed in the cell membrane of Xenopus oocytes. In the same expression system, untagged LvGLUT2 resulted to be a bidirectional glucose transporter that functions moving glucose down its concentration gradient in and out of the cell. Lvglut2 mRNA is expressed in hepatopancreas while in muscle and gills it was not detected. Hypoxia up-regulates the expression of Lvglut2 transcripts in hepatopancreas. These results provide a better understanding of facilitative glucose transporters and gene regulation during hypoxia in crustaceans.

White Spot Syndrome Virus Orf514 Encodes a Bona Fide DNA Polymerase

White spot syndrome virus (WSSV) is the causative agent of white spot syndrome, one of the most devastating diseases in shrimp aquaculture. The genome of WSSV includes a gene that encodes a putative family B DNA polymerase (ORF514), which is 16% identical in amino acid sequence to the Herpes virus 1 DNA polymerase. The aim of this work was to demonstrate the activity of the WSSV ORF514-encoded protein as a DNA polymerase and hence a putative antiviral target. A 3.5 kbp fragment encoding the conserved polymerase and exonuclease domains of ORF514 was overexpressed in bacteria. The recombinant protein showed polymerase activity but with very low level of processivity. Molecular modeling of the catalytic protein core encoded in ORF514 revealed a canonical polymerase fold. Amino acid sequence alignments of ORF514 indicate the presence of a putative PIP box, suggesting that the encoded putative DNA polymerase may use a host processivity factor for optimal activity. We postulate that WSSV ORF514 encodes a bona fide DNA polymerase that requires accessory proteins for activity and maybe target for drugs or compounds that inhibit viral DNA replication.

Status of the Yaqui Catfish (Ictalurus pricei) in the United States and Northwestern Mexico

Abstract To appraise conservation status of the Yaqui catfish Ictalurus pricei, we reviewed literature and unpublished records on a captive stock, examined voucher specimens at museums, re-sampled historical localities in the Yaqui, Mayo, and Fuerte river basins, and we surveyed rivers further south. A total of 72 specimens of native Ictalurus was collected in the Yaqui, Fuerte, Sinaloa, Culiacán, and San Lorenzo river basins. No native Ictalurus was collected in the Mayo Basin. Distribution of the Yaqui catfish appears restricted to the Yaqui, Mayo and Fuerte river basins, all of which now harbor nonnative blue (I. furcatus) and channel (I. punctatus) catfishes. The nonnative black bullhead (Ameiurus melas) is now known from the Yaqui Basin and the flathead catfish (Pylodictis olivaris) has been recorded anecdotally in the Yaqui Basin. Threats to the Yaqui catfish have increased in recent years and hybridization with the channel catfish now appears widespread. We conclude that the Yaqui catfish should be considered endangered throughout its range and that status of native populations of Ictalurus in the United States and Mexico should be reviewed and management intensified.