Identification and characterization of troponin genes in Locusta migratoria.

Troponin complex comprises three subunits, namely troponin C (TpnC), troponin I (TpnI) and troponin T (TpnT), and regulates the contraction of striated muscle. We found that the locust Locusta migratoria genome has one TpnT gene (LmTpnT), one TpnI gene (LmTpnI) and three TpnC genes (LmTpnC1, LmTpnC2 and LmTpnC3). Through alternative splicing, LmTpnT and LmTpnI potentially encode two and eight isoforms, respectively. The flight muscle and the jump muscle of L. migratoria express an identical LmTpnT isoform, but different LmTpnC isoforms and LmTpnI isoforms. LmTpnC2 and LmTpnC3 both contain highly conserved residues essential for calcium binding in the EF-hand II and IV, thus belonging two-site isoform. LmTpnC1 contains non-conserved substitutions in the EF-hand II and all highly conserved residues for calcium binding in the EF-hand IV. Mutagenesis and tyrosine fluorescence spectroscopic analysis show that both the EF-hand II and IV of LmTpnC1 can serve as calcium-binding site. Therefore, all three LmTpnC isoforms belong to two-site isoform. This is in contrast to the situation in the insect with asynchronous flight muscle, which expresses both one-site isoform and two-site isoform of TpnC. Those results suggest that the origination of insect asynchronous flight muscle is associated with the emergence of one-site isoform of TpnC.

Beneficial effects of Ectothiorhodospira shaposhnikovii WF on larval cultivation of Litopenaeus vannamei.

To develop high quality probiotics for shrimp larviculture, the effects of a photosynthetic purple sulphur bacterium WF identified as Ectothiorhodospira shaposhnikovii on survival and development of Litopenaeus vannamei larvae were evaluated in vivo. The larvae exhibited a better survival rate after administration of strain WF compared to the probiotic Rhodopseudomonas palustris. To investigate the effect of dose and dosing frequency, strain WF was added to larvae, stages nauplius 6 to zoea 3, at three different doses and dosing frequencies. Larval treatment with strain WF twice at 10(6) cfu/ml exhibited significantly higher survival compared to the other doses and dosing frequencies as well as the control. The effect on water quality was assessed by applying strain WF to larvae, stages nauplius 6 to postlarvae 1, under conditions of zero water exchange and one-third water exchange. The larvae exhibited higher survival and faster growth when treated under conditions of zero water exchange. No significant difference was detected in the levels of three water quality parameters and in vibrio counts between these two conditions. Therefore, E. shaposhnikovii WF acts both as a bioremediation agent and nutrient source and can benefit shrimp larvae if given at an appropriate dose and dosing frequency. Strain WF, a moderate halophile, shows great promise as a water additive in improving water quality and providing nutrition for shrimp larviculture.

Stability of a biological tissue fixed with a naturally occurring crosslinking agent (genipin).

The study was undertaken to investigate the stability of a biological tissue fixed with a naturally occurring crosslinking agent (genipin) at distinct elapsed storage durations. The glutaraldehyde-fixed counterpart was used as a control. Porcine pericardia procured from a slaughterhouse were used as raw materials. After fixation, the fixed tissues were sterilized in a graded series of ethanol solutions and thoroughly rinsed in phosphate buffered saline for 1 day, and then stored in a jar containing sterilized water. The samples were taken out and tested for their stability during the durations of 1day through 6 months after storage. The stability of each study group was tested by measuring its tensile strength, free-amino-group content, and denaturation temperature. Additionally, the cytotoxicity of each test sample and its corresponding storage solution were investigated in vitro using 3T3 fibroblasts. The results were examined using a microscope and 3-(4,5-dimethylthiazol-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. It was found that the stability of the genipin-fixed tissue during storage was superior to its glutaraldehyde-fixed counterpart. The differences in stability between the genipin- and glutaraldehyde-fixed tissues during storage may be caused by their differences in crosslinking structure. There was no apparent cytotoxicity for both the genipin-fixed tissue and its corresponding storage solution throughout the entire course of the study, whereas significant cytotoxicity was observed for both the glutaraldehyde-fixed tissue and its storage solution. However, the cytotoxicity of the glutaraldehyde-fixed tissue decreased with increasing elapsed storage duration, whereas that of its corresponding storage solution increased. This suggested that the toxic residues remaining in the glutaraldehyde-fixed tissue leached out slowly into its corresponding storage solution during the course of storage.