Project description:Comparison of pre and post metamorphis phases of cane toad Keywords: Developmental Stages Samples were harvested at 9, 18, 28 (pre-metamorph), 30 and 53 (post-metamorph) days of age and RNA extracted using Trizol Reagent (Invitrogen). Total RNA (100 mg) was reverse transcribed (Qiagen) and labelled cDNA probes were generated using the fluorophores, Cy3 and Cy5 (Amersham Pharmacia Biotech). Prehybridization of slides, application of the probe to the microarray slides, hybridisation, and subsequent washing steps were performed according to manufacturer's instructions (Corning Microarray Technology). Slides were scanned and analysed with a GenePix 4000B laser scanner and GenePix 3.0 and 4.0 software (Axon Instruments).
Project description:A ranavirus associated with mass mortalities in wild water frogs (Pelophylax spp.) and other amphibians in the Netherlands since 2010 was isolated, and its complete genome sequence was determined. The virus has a genome of 107,772 bp and shows 96.5% sequence identity with the common midwife toad virus from Spain.
Project description:Differential expression was determined in Calu-3 cells between mock infected and infected with H1N1 influenza virus A/Netherlands/602/2009 at nine time points post-infection. As a comparison, cells were also infected with A/CA/04/2009 H1N1 influenza virus at 4 time points post-infection.
Project description:Boreal toads (Anaxyrus boreas boreas) of the Southern Rocky Mountain population are declining due to the introduction of the chytrid fungus Batrachochytrium dendrobatidis (Bd). Boreal toads in Colorado are generally susceptible to Bd infection, but some Bd-tolerant populations persist in parts of the Southern Rocky Mountain and broader Eastern boreal toad population. We conducted a Bd challenge with lab-reared sibling toads from Bd-susceptible Colorado and purportedly Bd-tolerant Utah populations and report on transcriptomic responses to Bd during late infection in skin and liver tissue. Fewer immune genes were expressed in response to Bd in Colorado toads, but with greater upregulation compared to Utah toads, indicating a dysregulated immune response. Signatures of Bd-tolerance in Utah toads included more moderate upregulation in immune gene expression and a significantly enriched suite of gene functions related to innate and adaptive immune responses. Our transcriptomic results support the notion that Utah toads are tolerant to Bd, rather than resistant, carrying Bd loads similar to Colorado yet having a unique transcriptomic profile and presenting minimal clinical signs of chytridiomycosis. We conclude that closely related populations have divergent transcriptomic responses to Bd with a dysregulated immune response in Bd-susceptible toads.
Project description:Toad venom is a traditional Chinese medicine with high medicinal value. The existing quality evaluation standards of it has obvious limitation for lack of protein study. Thus, it’s necessary to screen suitable biomarkers and establish appropriate quality evaluation method for the characteristics of toad venom’s proteins, guaranteeing the safety and efficacy in clinical applications. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis, high-performance liquid chromatography and cytotoxicity assay were used to analyze protein components difference between toad venom of different areas. Functional proteins were screened as potential biomarkers by tandem mass tags proteomic and bioinformatic analysis. The results showed that the composition of different toad venom was similar. The protein content of Lianyungang’s toad venom was higher and the alcohol-soluble substance was lower, which suggested that they had no correlation. Simultaneously, the protein component was verified to have strong cytotoxicity. Proteomics analysis showed that 13 antimicrobial proteins, 4 anti-inflammatory and analgesic proteins, and 20 antitumor proteins were screened in the extracellular differentially expressed proteins. A candidate list of functional proteins that could be used as potential biomarkers was presented. Moreover, the antimicrobial activity of Lysozyme C-1 and the anti-inflammatory and analgesic activity of Neuropeptide B were identified as biomarkers for toad venom protein in future. The biomarkers can form the basis for the quality studies of toad venom’s proteins, helping construct and improve the safe, scientific and comprehensive quality evaluation method.