Document Type

Theses, Ph.D

Rights

This item is available under a Creative Commons License for non-commercial use only

Disciplines

1.6 BIOLOGICAL SCIENCES

Publication Details

Successfully submitted for the award of Doctor of Philosophy (Ph.D) to the Dublin Institute of Technology, 2006.

Abstract

The aims of this research project were to study the biology of abalone through a histological characterisation of two species of this shellfish, Haliotis tuberculata and Haliotis discus hannai using both histochemical and immunohistochemical techniques and to develop molecular methods for the detection and quantitation of common shellfish pathogens associated with abalone disease outbreaks. In the first part of this study immunohistochemistry was used to examine cell distribution structural and functional proteins in abalone tissues. Phenotypic antigen expression was analysed in abalone shellfish using an array of antibodies that react with a range of proteins in many different species. In the second section of this study abalone functional biochemistry was characterised using both histochemical and enzyme histochemical staining methodologies. The distribution of both functional and structural elements such as enzymes, carbohydrates, lipids, pigments and minerals was investigated in abalone tissues. The potential of infectious pathogens to cause high mortalities within shellfish population has led to increased awareness of shellfish disease, along with the development of methods for the detection of pathogenic organisms. Vibrio harveyi (bacterium) and Perkinsus olseni (protozoan) are two major disease causing pathogens of abalone. In the final part of this study, nucleic acid-based methods for the detection and quantitation of V. harveyi and P. olseni were developed. A polymerase chain reaction (PCR) method was developed to detect DNA from isolates of V. harveyi and further developed into a novel multiplex PCR method, which simultaneously amplifies a 413 bp region of the 16S rRNA gene of V. harveyi and a 155 bp region of the actin mRNA gene of Haliotis spp. A PCR amplification method to detect P. olseni using novel primers (designed in this study) which amplify a 265 bp DNA fragment of the ITS region of the genome of this organism was also developed. Both PCR methods were optimised for pathogen detection in alcohol preserved and paraffin embedded tissue samples. For rapid detection and quantitation of V. harveyi and P. olseni, two real-time SYBR Green PCR methods were developed.

DOI

10.21427/D72K5K

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