Lai, Sze Tsai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 94-104). / Abstracts in English and Chinese. / Thesis Committees --- p.i / Statement --- p.ii / Abstract --- p.iii / 摘要 --- p.v / Acknowledgments --- p.vi / List of Abbreviations --- p.vii / Table of Contents --- p.ix / List of Tables --- p.xiii / List of Figures --- p.xiv / Chapter 1 --- Literature review --- p.1 / Chapter 1.1 --- Introduction to Cordyceps --- p.1 / Chapter 1.2 --- Fungal proteomics --- p.2 / Chapter 1.2.1 --- Extraction method --- p.2 / Chapter 1.2.2 --- Proteomic study of Cordyceps --- p.3 / Chapter 1.3 --- Ingredients of Cordyceps and their related biological activities --- p.5 / Chapter 1.3.1 --- Polysaccharides --- p.5 / Chapter 1.3.2 --- Nucleosides --- p.6 / Chapter 1.3.2.1 --- Cordycepin --- p.6 / Chapter 1.3.2.2 --- Adenosine --- p.8 / Chapter 1.4 --- Cordyceps and their related biological activities --- p.9 / Chapter 1.4.1 --- Cordyceps militaris --- p.9 / Chapter 1.4.2 --- Cordyceps sinensis --- p.10 / Chapter 1.5 --- Proteomic analysis of proteome change --- p.12 / Chapter 1.5.1 --- Proteomic tools used to study the change in protein expression --- p.12 / Chapter 1.5.2 --- Two-dimensional gel electrophoresis --- p.13 / Chapter 1.5.3 --- Mass spectrometry --- p.13 / Chapter 1.6 --- Objective --- p.16 / Chapter 2 --- Methodology --- p.17 / Chapter 2.1 --- Cultivation of Cordyceps militaris --- p.17 / Chapter 2.2 --- Proteomic study on Cordyceps militaris --- p.17 / Chapter 2.2.1 --- Extraction of proteins from Cordyceps militaris --- p.17 / Chapter 2.2.2 --- Protein quantification --- p.18 / Chapter 2.2.3 --- 2D Gel electrophoresis --- p.19 / Chapter 2.2.4 --- Image analysis --- p.20 / Chapter 2.2.5 --- In gel digestion and MALDI-ToF MS --- p.20 / Chapter 2.3 --- Preparation of Cordyceps extracts for anti-proliferation assay on cell lines --- p.21 / Chapter 2.3.1 --- Types of the extracts of Cordyceps --- p.21 / Chapter 2.3.2 --- Preparation of the extracts of Cordyceps --- p.21 / Chapter 2.4 --- Anti-proliferation assay on cell lines for extract screening --- p.22 / Chapter 2.4.1 --- Cell lines and culturing condition --- p.22 / Chapter 2.4.2 --- Viable cell count using trypan blue exclusion method --- p.22 / Chapter 2.4.3 --- Anti-proliferation assay on SV7 tert using MTT assay --- p.23 / Chapter 2.4.4 --- Determination of the IC5o values --- p.24 / Chapter 2.4.5 --- Statistical Analysis --- p.24 / Chapter 2.5 --- Anti-proliferation assay on other cell lines using the two screened extracts --- p.24 / Chapter 2.5.1 --- Cell lines and culturing condition --- p.24 / Chapter 2.5.2 --- "Anti-proliferation assay on on HepG2, H292, Neuro2a，WIL2-NS cells using MTT assay" --- p.25 / Chapter 2.6 --- Proteomic studies for SV7tert and Hs68 after the treatment of extracts --- p.25 / Chapter 2.6.1 --- Protein sample preparation of SV7tert and Hs68 --- p.25 / Chapter 2.6.2 --- Protein quantification --- p.26 / Chapter 2.6.3 --- 2D Gel electrophoresis --- p.26 / Chapter 2.6.4 --- Image analysis --- p.26 / Chapter 2.7 --- Western Immunoblotting --- p.26 / Chapter 2.7.1 --- Protein sample preparation of SV7tert and Hs68 --- p.26 / Chapter 2.7.2 --- SDS-PAGE --- p.27 / Chapter 2.7.3 --- Protein Blotting --- p.27 / Chapter 2.7.4 --- Membrane Blocking and Antibody Incubations --- p.28 / Chapter 2.7.5 --- Detection of Proteins --- p.28 / Chapter 3 --- Results --- p.29 / Chapter 3.1 --- Proteins identification in Cordyceps militaris --- p.29 / Chapter 3.1.1 --- 2D gel electrophoresis analysis and resolution --- p.29 / Chapter 3.1.2 --- Identification and categorization of proteins of mycelia and fruiting body of Cordyceps militaris --- p.30 / Chapter 3.2 --- Investigation of anti-proliferating activity of extracts using MTT assays on SV7tert and Hs68 cell lines --- p.44 / Chapter 3.2.1 --- Mycelia extract from Cordyceps militaris --- p.44 / Chapter 3.2.2 --- Fruiting body extract from Cordyceps militaris --- p.44 / Chapter 3.2.3 --- Mycelia extract from Cordyceps sinensis --- p.47 / Chapter 3.2.4 --- Fruiting body extract from Cordyceps sinensis --- p.47 / Chapter 3.2.5 --- Screening of extracts --- p.50 / Chapter 3.3 --- "Investigation of anti-proliferating activity of extracts using MTT assays on HepG2，H292, Neuro2a and WIL2-NS cell lines" --- p.51 / Chapter 3.3.1 --- Mycelia extract from Cordyceps militaris --- p.51 / Chapter 3.3.2 --- Fruiting body extract from Cordyceps militaris --- p.51 / Chapter 3.4 --- Changes in total protein expression profiles in SV7tert and Hs68 cell lines --- p.56 / Chapter 3.4.1 --- Corresponding extract treatment of cell lines --- p.56 / Chapter 3.4.2 --- 2D gel electrophoresis analysis of protein from cells (SV7tert or Hs68) --- p.56 / Chapter 3.4.2.1 --- SV7tert study --- p.57 / Chapter 3.4.2.2 --- Hs68 study --- p.57 / Chapter 3.4.3 --- Protein identification --- p.65 / Chapter 3.4.3.1 --- Changes in protein expressions in SV7tert after mycelia extract treatment --- p.65 / Chapter 3.4.3.2 --- Changes in protein expressions in Hs68 after mycelia extract treatment --- p.65 / Chapter 3.4.3.3 --- Changes in protein expressions in SV7tert after fruiting body extract treatment --- p.66 / Chapter 3.4.3.4 --- Changes in protein expressions in Hs68 after fruiting body extract treatment --- p.66 / Chapter 3.5 --- Western immunoblotting --- p.71 / Chapter 3.5.1 --- Corresponding extract treatment of cell lines --- p.71 / Chapter 3.5.2 --- Normalization of protein loading using anti-actin antibody --- p.73 / Chapter 3.5.3 --- Detection of caspase 3 by use of antibody --- p.74 / Chapter 3.5.4 --- Detection of cleaved caspase 3 by use of antibody --- p.74 / Chapter 4 --- Discussion --- p.77 / Chapter 4.1 --- Identification of proteins in Cordyceps militaris --- p.77 / Chapter 4.2 --- Difficulties in identifying the proteins in Cordyceps militaris --- p.80 / Chapter 4.3 --- Investigation of anti-proliferating activity of extracts --- p.80 / Chapter 4.4 --- Changes in cell total protein expression profiles in SV7tert and Hs68 cell lines --- p.81 / Chapter 4.4.1 --- Protein alterations in SV7tert treated with mycelia extract --- p.82 / Chapter 4.4.1.1 --- Heat shock protein 27 (Hsp27) --- p.82 / Chapter 4.4.1.2 --- Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) --- p.83 / Chapter 4.4.2 --- Protein alterations in Hs68 with mycelia extract treatment --- p.84 / Chapter 4.4.2.1 --- Chain B of triosephosphate isomerase - Triosephophate isomerase 1 --- p.84 / Chapter 4.4.2.2 --- Glutathione transferase --- p.85 / Chapter 4.4.3 --- Protein alterations in SV7tert with fruiting body extract treatment --- p.86 / Chapter 4.4.3.1 --- Calreticulin precusor --- p.86 / Chapter 4.4.3.2 --- Nucleophosmin 1 isoform 2 (B23) --- p.87 / Chapter 4.4.3.3 --- Heat shock 70kDa protein 8 isoform 1 - Heat shock 70kDa protein (Hsp70) --- p.88 / Chapter 4.4.3.4 --- Voltage-dependent anion channel 2 (VDAC2) --- p.89 / Chapter 4.4.3.5 --- "Tumor protein, translationally controlled (TCTP)" --- p.90 / Chapter 4.4.3.6 --- RAN binding protein 1 (RANBP1) --- p.91 / Chapter 4.4.4 --- Protein alteration in Hs68 with mycelia extract treatment --- p.92 / Chapter 4.5 --- Conclusion --- p.93 / References --- p.94

No description available.

Graduation date: 2006

According to traditional Chinese medicine , Cordyceps sinensis (CS) can prevent subjects from renal failure. The aim of this study was to investigate the protective effect of CS preconditioning on ischemic renal acute failure in rats and to assess its mechanism. The animal model of ischemic acute renal failure was performed by left nephrectomy and clamping right renal vessel for 45 mins in S-D rats. Cordyceps group had been pretreated with two-day 600 mg/kg CS before I/R injury. Rats were sacrificed at 1, 3, 6, 16, 48 and 120 h after reperfusion for evaluation of renal function and histopathological PASD staining. The immunohistochemistry and Western blotting of SDF-1£\, CXCR4 and Ki67 were also performed. £E-galactosidase activity was detected with the senescence staining. The results showed that the level of creatinine in Cordyceps group were significant lower after 48 hours I/R injury (p =0.04). PSAD staining in Cordyceps group revealed less tubular necrosis, tubular dilatation, and cast formation at 6 and 16 hour than in control group. Immunohistochemistry of SDF-1£\ in Cordyceps group demonstrated staining in the distal tubules and collecting ducts at 1, 3, 6, and 16 h. The CXCR4 signal of control group had gradually intensified from 1 to 6 hr after I/R . In Cordyceps group, the CXCR4 expression had been stabilized until 16 h after I/R. The £]-galactosidase activity was higher in control group at 1, 3 and 6 hours. However, the senescence was presented at 1 and 3 hours in Cordyceps group. The nuclear staining of repair enzyme Ki67 in Cordyceps group showed higher density than in control group. Pathologic morphology indicated CS may protect subjects from ischemic acute renal failure. CS also induced SDF-1£\ expression in early stage of I/R injury, and maintained the stable CXCR4 expression. CS can not only reduce the activity of senescence-related £]-galactosidase, but also regulate the expression of repair enzyme Ki67, indicating that CS may alleviate the ischemic-induced senescence and enhance renal repair.

Fung, Chun Kit. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 121-135). / Abstracts in English and Chinese. / Declaration --- p.i / Acknowledgement --- p.ii / Abbreviations --- p.iii / Abstract in English --- p.v / Abstract in Chinese --- p.vii / Table of Contents --- p.ix / List of Figures --- p.xii / Chapter Chapter 1 - --- Introduction / Chapter 1.1 --- Genus Cordyceps --- p.1 / Chapter 1.2 --- Cordyceps militaris --- p.4 / Chapter 1.3 --- Biological Functions and Chemical Constituents of Cordyceps militaris --- p.9 / Chapter 1.4 --- "Human Bronchial Epithelial Cell Line, 16HBE14o-" --- p.13 / Chapter 1.5 --- Ion Transport in Human Bronchial Epithelial Cells --- p.16 / Chapter 1.6 --- Objectives of the Experiments --- p.20 / Chapter Chapter 2 - --- Materials and Methods / Chapter 2.1 --- Solutions and Chemicals --- p.21 / Chapter 2.2 --- Preparation of Hot Water Cordyceps militaris Extract --- p.22 / Chapter 2.3 --- Culture of Cells --- p.23 / Chapter 2.4 --- Short-Circuit Current (lsc) Measurement --- p.24 / Chapter 2.5 --- Short-Circuit Current (lsc) Measurement in Nystatin-Permeabilized Monolayer --- p.29 / Chapter 2.6 --- Measurements of [Ca2+]i --- p.31 / Chapter 2.7 --- Measurement of PKA Activity --- p.36 / Chapter 2.8 --- Statistical Analysis --- p.37 / Chapter Chapter 3 - --- Results / Chapter 3.1 --- Regulation of Ion Transport in 16HBE14o- Cells by CM Water Extract --- p.38 / Chapter 3.1.1 --- Dose-Dependent Relationship of CM Water Extract --- p.39 / Chapter 3.1.2 --- "Involvement of CI"" Transport in CM-induced lsc Response" --- p.42 / Chapter 3.1.3 --- Involvement of K+ channels in CM-induced lsc Response --- p.47 / Chapter 3.1.4 --- Involvement of Adenylate Cyclase/cAMP/Protein Kinase A Pathway in CM-induced lsc Response --- p.52 / Chapter 3.1.5 --- Involvement of Ca2+-Dependent Pathway in CM-induced lsc Response --- p.57 / Chapter 3.1.6 --- "Effect of CM Extract on Apical CI"" Current and Basolateral K+ Current in Nystatin-Permeabilized Epithelia" --- p.61 / Chapter 3.1.7 --- Effect of CM Extract on PKA Activity --- p.67 / Chapter 3.2 --- Regulation of Ion Transport in 16HBE14o- Cells by Cordycepin --- p.70 / Chapter 3.2.1 --- Dose-Dependent Relationship of Cordycepin --- p.70 / Chapter 3.2.2 --- "Involvement of CI"" Transport in Cordycepin-induced lsc Response" --- p.73 / Chapter 3.2.3 --- Involvement of K+ channels in Cordycepin-induced lsc Response --- p.79 / Chapter 3.2.4 --- Involvement of Adenylate Cyclase/cAMP/Protein Kinase A Pathway in Cordycepin-induced lsc Response --- p.84 / Chapter 3.2.5 --- Involvement of Ca2+-Dependent Pathway in Cordycepin-induced lsc Response --- p.89 / Chapter 3.2.6 --- Effect of Cordycepin on Intracellular Ca2+ Concentrations --- p.93 / Chapter 3.2.7 --- "Effect of Cordycepin on Apical CI"" Current and Basolateral K+ Current in Nystatin-Permeabilized Epithelia" --- p.98 / Chapter 3.2.8 --- Effect of Cordycepin on PKA Activity --- p.104 / Chapter Chapter 4 - --- Discussion / Chapter 4.1 --- Regulation of Ion Transport in 16HBE14o- Cells by CM Extract --- p.107 / Chapter 4.2 --- Intracellular Signaling Mechanisms behind CM-induced lsc Responses --- p.110 / Chapter 4.3 --- Regulation of Ion Transport in 16HBE14o- Cells by Cordycepin --- p.111 / Chapter 4.4 --- Intracellular Signaling Mechanisms behind Cordycepin-induced lsc Responses --- p.114 / Chapter Chapter 5 - --- Conclusion / Chapter 5.1 --- Summary --- p.117 / Chapter 5.2 --- Future Directions --- p.120 / Chapter Chapter 6 - --- References --- p.121 / Chapter Chapter 7 - --- Publications --- p.136

Chan, Wing Hin. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 126-143). / Abstracts in English and Chinese. / Acknowledgement --- p.ii / Abstract --- p.iii / 摘要 --- p.vi / Table of Content --- p.viii / List of Figures --- p.xiv / List of Tables --- p.xvi / Abbreviations --- p.xviii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Food and herb authentication --- p.1 / Chapter 1.1.1 --- Background and definition --- p.1 / Chapter 1.1.2 --- Importance of species identification in food and herb authentication --- p.2 / Chapter 1.1.2.1 --- Primary health care --- p.2 / Chapter 1.1.2.2 --- Food and herb safety --- p.3 / Chapter 1.1.2.3 --- Conservation --- p.4 / Chapter 1.1.3 --- Methods for species identification in food and herb authentication --- p.4 / Chapter 1.1.3.1 --- Morphological identification --- p.5 / Chapter 1.1.3.2 --- Chemical analysis --- p.6 / Chapter 1.1.3.3 --- Molecular analysis --- p.9 / Chapter 1.1.4 --- Legislation --- p.11 / Chapter 1.1.4.1 --- Labeling ´ب --- p.11 / Chapter 1.1.4.2 --- Chinese medicine : --- p.12 / Chapter 1.1.4.3 --- Conservation --- p.12 / Chapter 1.2 --- Dongchongxiacao --- p.13 / Chapter 1.2.1 --- Background information of Dongchongxiacao --- p.13 / Chapter 1.2.2 --- Classification of fungal part of Dongchongxiacao --- p.14 / Chapter 1.2.3 --- Dongchongxiacao as a Traditional Chinese Medicine. --- p.15 / Chapter 1.2.4 --- The Dongchongxiacao market --- p.16 / Chapter 1.2.5 --- Adulteration and contamination of Dongchongxiacao --- p.18 / Chapter 1.2.6 --- Authentication of Dongchongxiacao --- p.19 / Chapter 1.2.6.1 --- Morphological identification --- p.19 / Chapter 1.2.6.2 --- Chemical analysis --- p.20 / Chapter 1.2.6.3 --- Molecular analysis --- p.22 / Chapter 1.2.6.3.1 --- "FINS analysis with genomic ITS, nrLSU, EF-lα and rpbl regions for fungal analyses" --- p.22 / Chapter 1.2.6.3.2 --- FINS analysis with mitochondrial CytB and COI regions for caterpillar analyses --- p.24 / Chapter 1.3 --- Abalone --- p.26 / Chapter 1.3.1 --- Background information of abalone --- p.26 / Chapter 1.3.2 --- Abalone as food --- p.27 / Chapter 1.3.3 --- The abalone market --- p.28 / Chapter 1.3.4 --- Adulteration of abalone --- p.31 / Chapter 1.3.5 --- Authentication of abalone --- p.32 / Chapter 1.3.5.1 --- Morphological identification --- p.32 / Chapter 1.3.5.2 --- Chemical analysis --- p.32 / Chapter 1.3.5.3 --- Molecular analysis --- p.33 / Chapter 1.3.5.3.1 --- FINS analysis with mitochondrial COI and 16S rDNA --- p.33 / Chapter 1.3.5.3.2 --- Haliotis-specific detection --- p.34 / Chapter 1.4 --- Aim and Objectives --- p.35 / Chapter Chapter 2 --- Materials and Methods --- p.36 / Chapter 2.1 --- Materials used in this sutdy --- p.36 / Chapter 2.1.1 --- Dongchongxiacao and Cordyceps samples --- p.36 / Chapter 2.1.2 --- Downloaded sequences from NCBI database included in Dongchongxiacao study. --- p.45 / Chapter 2.1.3 --- Abalone and gastropod samples --- p.48 / Chapter 2.1.4 --- Downloaded sequences from NCBI database included in abalone study --- p.54 / Chapter 2.2 --- Reagents and equipments : --- p.56 / Chapter 2.2.1 --- Chemical test on the presence of potassium alum in Dongchongxiacao --- p.56 / Chapter 2.2.2 --- Sample preparation and DNA extraction --- p.57 / Chapter 2.2.3 --- Polymerase Chain Reaction --- p.57 / Chapter 2.2.4 --- Agarose gel electrophoresis and Gene Clean --- p.57 / Chapter 2.2.5 --- Cloning --- p.58 / Chapter 2.2.6 --- Cycle sequencing --- p.58 / Chapter 2.3 --- Experimental procedures --- p.58 / Chapter 2.3.1 --- Morphological observation of Dongchongxiacao and abalone --- p.59 / Chapter 2.3.2 --- Chemical test of potassium in Dongchongxiacao --- p.59 / Chapter 2.3.3 --- Sample preparation and DNA extraction --- p.60 / Chapter 2.3.4 --- Polymerase Chain Reaction --- p.61 / Chapter 2.3.5 --- Agarose gel electrophoresis and Gene Clean --- p.64 / Chapter 2.3.6 --- Cloning --- p.65 / Chapter 2.3.7 --- Cycle sequencing --- p.67 / Chapter 2.3.8 --- Sequence analyses --- p.67 / Chapter 2.3.9 --- Haliotis-specific primer design and PCR test --- p.68 / Chapter Chapter 3 --- Results --- p.71 / Chapter 3.1 --- Dongchongxiacao --- p.71 / Chapter 3.1.1 --- Morphological observations --- p.71 / Chapter 3.1.2 --- Chemical test of potassium alum --- p.77 / Chapter 3.1.3 --- Sequence analyses --- p.79 / Chapter 3.1.4 --- The dendrograms --- p.81 / Chapter 3.2 --- Abalone --- p.91 / Chapter 3.2.1 --- Morphological observations --- p.91 / Chapter 3.2.2 --- Sequence analyses --- p.92 / Chapter 3.2.3 --- The dendrograms --- p.94 / Chapter 3.2.4 --- Haliotis-specific PCR --- p.96 / Chapter Chapter 4 --- Discussion --- p.98 / Chapter 4.1 --- Dongchongxiacao --- p.98 / Chapter 4.1.1 --- Species identification of Dongchongxiacao and related Cordyceps species --- p.98 / Chapter 4.1.1.1 --- Ophiocordyceps sinensis --- p.98 / Chapter 4.1.1.2 --- Cordyceps gunnii --- p.100 / Chapter 4.1.1.3 --- Metacordyceps taii --- p.102 / Chapter 4.1.1.4 --- Cordyceps militaris --- p.103 / Chapter 4.1.2 --- Adulteration of Dongchongxiacao and labeling --- p.104 / Chapter 4.1.3 --- Hosts of Dongchongxiacao fungi and relationship between them --- p.107 / Chapter 4.2 --- Abalone --- p.109 / Chapter 4.2.1 --- Species identification of abalones and other gastropod species by FINS analysis --- p.109 / Chapter 4.2.1.1 --- Haliotis species --- p.109 / Chapter 4.2.1.1.1 --- Haliotis diversicolor --- p.110 / Chapter 4.2.1.1.2 --- Haliotis discus --- p.110 / Chapter 4.2.1.1.3 --- Haliotis asinina --- p.111 / Chapter 4.2.1.1.4 --- Haliotis rufescens --- p.111 / Chapter 4.2.1.1.5 --- Haliotis midae --- p.111 / Chapter 4.2.1.1.6 --- Haliotis madaka --- p.112 / Chapter 4.2.1.1.7 --- Haliotis rubra --- p.113 / Chapter 4.2.1.1.8 --- Haliotis iris --- p.113 / Chapter 4.2.1.1.9 --- Haliotis corrugata --- p.114 / Chapter 4.2.1.2 --- Concholepas concholepas --- p.114 / Chapter 4.2.1.3 --- Hemifusus species --- p.115 / Chapter 4.2.1.4 --- """Dried abalone slice"" samples (D1 to D3) and canned top-shell (E5)" --- p.115 / Chapter 4.2.2 --- Haliotis-speciflc PCR --- p.115 / Chapter 4.2.3 --- Adulteration of abalone and labeling --- p.116 / Chapter 4.3 --- Significance and limitation of molecular approaches in authentication of food and herbs --- p.117 / Chapter 4.3.1 --- FINS analysis --- p.117 / Chapter 4.3.1.1 --- High interspecific variability but low intraspecific variations --- p.118 / Chapter 4.3.1.2 --- Amplification with universal primers --- p.118 / Chapter 4.3.1.3 --- Insufficient DNA sequence available in database --- p.119 / Chapter 4.3.1.4 --- Contamination by foreign DNA and amplification of undesirable DNA in sample mixture --- p.120 / Chapter 4.3.1.5 --- Amplification of degraded DNA --- p.121 / Chapter 4.3.1.6 --- Suggested regions for authentication of Dongchongxiacao and abalone based on FINS analysis results --- p.121 / Chapter 4.3.2 --- PCR with specific primers for targeted amplicons --- p.122 / Chapter 4.3.3 --- Other limitations of molecular approaches in authentication of food and herbs --- p.123 / Chapter 4.4 --- Further investigation --- p.124 / Chapter 4.5 --- Conclusion --- p.124 / References : --- p.126 / Chapter Appendix 1 --- Sequence alignment of 16S rDNA gene sequences of abalone for Haliotis-specific primer design --- p.144 / Chapter Appendix 2 --- Accession numbers of sequences of Dongchongxiacao and Cordyceps samples in this study --- p.149 / Chapter Appendix 3 --- Search results of CytB sequences of caterpillar host of Cordyceps samples based on BLAST search results from GenBank --- p.150 / Chapter Appendix 4 --- Search results of COI sequences of caterpillar host of Cordyceps samples based on BLAST search results from GenBank --- p.151 / Chapter Appendix 5 --- Search results of COI sequences of caterpillar host of Cordyceps samples based on BLAST search results from GenBank --- p.152 / Chapter Appendix 6 --- Sequence alignment of ITS sequences of Cordyceps samples and related sequences --- p.153 / Chapter Appendix 7 --- Sequence alignment of nrLSU sequences of Cordyceps samples and related sequences --- p.161 / Chapter Appendix 8 --- Sequence alignment of EF-lα sequences of Cordyceps samples and related sequences --- p.168 / Chapter Appendix 9 --- Sequence alignment of rpbl sequences of Cordyceps samples and related sequences --- p.173 / Chapter Appendix 10 --- "Sequence alignment of combined dataset of three regions (nrLSU, EF-lα and rpbl) of Cordyceps samples and related sequences" --- p.179 / Chapter Appendix 11 --- Sequences alignment of CytB sequences of caterpillar host of Cordyceps samples and related sequences --- p.188 / Chapter Appendix 12 --- Sequence alignment of COI sequences of caterpillar host of Cordyceps samples and related sequences --- p.191 / Chapter Appendix 13 --- Sequence alignment of COI sequences of Cordyceps samples D12-2 and D14 and related sequences --- p.195 / Chapter Appendix 14 --- Sequence distance matrix of ITS sequences of Cordyceps samples and related samples based on K2P algorithm --- p.196 / Chapter Appendix 15 --- Sequence distance matrix of nrLSU sequences of Cordyceps samples and related samples based on K2P algorithm --- p.203 / Chapter Appendix 16 --- Sequence distance matrix of EF-lα sequences of Cordyceps samples and related samples based on K2P algorithm --- p.208 / Chapter Appendix 17 --- Sequence distance matrix of rpbl sequences of Cordyceps samples and related samples based on K2P algorithm --- p.213 / Chapter Appendix 18 --- "Sequence distance matrix of combined dataset of three regions (nrLSU, EF-lα and rpbl) sequences of Cordyceps samples and related samples based on K2P algorithm" --- p.217 / Chapter Appendix 19 --- Sequence distance matrix of CytB sequences of caterpillar host of Cordyceps samples and related samples based on K2P algorithm --- p.219 / Chapter Appendix 20 --- Sequence distance matrix of COI sequences of caterpillar host of Cordyceps samples and related samples based on K2P algorithm --- p.223 / Chapter Appendix 21 --- Sequence alignment of chloroplast trnH-psbA sequences of Cordyceps sample D12-2 and related sequences --- p.226 / Chapter Appendix 22 --- Accession numbers of sequences of abalone and gastropod samples in this study --- p.227 / Chapter Appendix 23 --- Search results of 16S rDNA sequences of the abalone and gastropod samples based on BLAST search results from GenBank --- p.228 / Chapter Appendix 24 --- Search results of COI sequences of the abalone and gastropod samples based on BLAST search results from GenBank --- p.229 / Chapter Appendix 25 --- Search results of COI sequences of the abalone and gastropod samples based on BOLD-IDS --- p.230 / Chapter Appendix 26 --- Sequence alignment of 16S sequences of abalone samples and related sequences --- p.231 / Chapter Appendix 27 --- Sequence alignment of COI sequences of abalone samples and related sequences --- p.234 / Chapter Appendix 28 --- Sequence alignment of COI sequences of abalone product sample D2 and related sequences --- p.238 / Chapter Appendix 29 --- Sequence distance matrix of 16S sequences of abalone samples and related samples based on K2P algorithm --- p.239 / Chapter Appendix 30 --- Sequence distance matrix of COI sequences of abalone samples and related samples based on K2P algorithm --- p.243

No description available.

The fungus Ophiocordyceps camponoti-floridani (Ophcf) infects Camponotus floridanus carpenter ants and manipulates them to climb to a high tree branch, bite down on foliage and die. Post ant death, the fungus grows out of the ant and spreads spores for reproduction. I investigated the function of an Ophcf gene product highly activated during the behavioral manipulation of these “zombie ants”; an enterotoxin. I have created an expression vector and heterologously expressed this enterotoxin in Cordyceps bassiana (Cbass), a related fungus that does not normally manipulate behavior. This process includes gene amplification, Golden Gate vector cloning in E. coli, A. tumefaciens-mediated transformation to Cbass, and RT-qPCR to verify heterologous gene expression. This was followed by carpenter ant infections with transgenic Cbass (EntB), wildtype Cbass (infected control), and sham (non-infected control) infected ants. Subsequent behavioral observations using tracking system MARGO (Werkhoven et al., 2019) have detected changes in activity levels of ants infected with both transgenic and WT Cbass compared to sham infected ants. This supports previously qualitative descriptions of increased activity caused during infection with WT Cbass (Trinh, 2020). There is a slight but insignificantly higher activity response from EntB compared to WT infected ants over the course of the trial that may be indicative of Ophcf induced changes that are different from general sickness behavior. Additional replicates are necessary to discern if these findings are statistically robust. Future studies should administer this enterotoxin expressing Cbass to observe inter-social behaviors of Carpenter ants. If the enterotoxin is sufficient to elicit one of the side effects of typical Ophcf infection, this would justify further characterization of the proteins and their functions in altering C. floridanus behavior. This characterization could yield information applicable to other parasite-host relationships as well.

University of Macau / Institute of Chinese Medical Sciences

University of Macau / Institute of Chinese Medical Sciences