The Growth and Pathogenesis of the Intracellular Pathogen, Piscirickettsia salmonis
Piscirickettsia salmonis is the causative agent of piscirickettsiosis or salmon rickettsia syndrome (SRS). It is a Gram-negative intracellular bacterium that causes significant mortalities to salmonids, particularly in Chile. The initial isolation and identification of P. salmonis occurred in fish c...
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ftmaineuniv:oai:digitalcommons.library.umaine.edu:etd-3589 2023-06-11T04:10:22+02:00 The Growth and Pathogenesis of the Intracellular Pathogen, Piscirickettsia salmonis Makrinos, Daniel 2016-12-01T08:00:00Z application/pdf https://digitalcommons.library.umaine.edu/etd/2552 https://digitalcommons.library.umaine.edu/context/etd/article/3589/viewcontent/Daniel_L_Makrinos_Final_Thesis_P._salmonis_1_10_17.pdf unknown DigitalCommons@UMaine https://digitalcommons.library.umaine.edu/etd/2552 https://digitalcommons.library.umaine.edu/context/etd/article/3589/viewcontent/Daniel_L_Makrinos_Final_Thesis_P._salmonis_1_10_17.pdf Electronic Theses and Dissertations Piscirickettsia salmonis agar growth intracellular salmon SRS Aquaculture and Fisheries Bacteriology Cell Biology Environmental Microbiology and Microbial Ecology Immunity Immunology of Infectious Disease Immunopathology Immunoprophylaxis and Therapy Marine Biology Medicinal Chemistry and Pharmaceutics Microbial Physiology Molecular Biology Organismal Biological Physiology Pathogenic Microbiology text 2016 ftmaineuniv 2023-05-04T18:02:37Z Piscirickettsia salmonis is the causative agent of piscirickettsiosis or salmon rickettsia syndrome (SRS). It is a Gram-negative intracellular bacterium that causes significant mortalities to salmonids, particularly in Chile. The initial isolation and identification of P. salmonis occurred in fish cell lines, therefore growth of P. salmonis is most often carried out in tissue culture. Three cell lines were analyzed in this study to determine which in vitro cellular environment was most suitable for additional research. Tissue culture has been used for the growth and cultivation of P. salmonis for nearly two decades, until the facultative nature of the pathogen was confirmed upon the development of blood and cysteine based agar. Since then, research has continued to drive the creation of novel agar and broth formulations in order to improve the efficacy of cultivation of P. salmonis in cell-free environments. In this thesis, several published agar and broth formulations were tested to understand how this intracellular bacterium grows in these environments. Cell-free media compositions were investigated by using previous formulations and atomic emission spectrophotometry in order to create novel media. This led to the development of MAK broth, MAK2 agar, and BCYE-SRS agar, which significantly improved overall cell-free growth. Furthermore, growth was evaluated using qPCR, SDS-PAGE gels, growth curves, cell counts, colony forming units, and TCID50 titration assays. Additionally, diagnosis of the pathogen was improved by the use of novel cell-free media, vital stains, and PCR assays. In order to study P. salmonis in vivo, two studies were conducted. The first used an intraperitoneal chamber model in Atlantic salmon to observe how the bacterium and host responded and intercommunicated among one another. A lethal dose trial was then carried out in freshwater to determine if P. salmonis grown in cell-free media was a viable option for future challenge trials. Overall, in vitro and in vivo studies provided an ... Text Atlantic salmon The University of Maine: DigitalCommons@UMaine Mak ENVELOPE(162.381,162.381,56.401,56.401) |
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Open Polar |
collection |
The University of Maine: DigitalCommons@UMaine |
op_collection_id |
ftmaineuniv |
language |
unknown |
topic |
Piscirickettsia salmonis agar growth intracellular salmon SRS Aquaculture and Fisheries Bacteriology Cell Biology Environmental Microbiology and Microbial Ecology Immunity Immunology of Infectious Disease Immunopathology Immunoprophylaxis and Therapy Marine Biology Medicinal Chemistry and Pharmaceutics Microbial Physiology Molecular Biology Organismal Biological Physiology Pathogenic Microbiology |
spellingShingle |
Piscirickettsia salmonis agar growth intracellular salmon SRS Aquaculture and Fisheries Bacteriology Cell Biology Environmental Microbiology and Microbial Ecology Immunity Immunology of Infectious Disease Immunopathology Immunoprophylaxis and Therapy Marine Biology Medicinal Chemistry and Pharmaceutics Microbial Physiology Molecular Biology Organismal Biological Physiology Pathogenic Microbiology Makrinos, Daniel The Growth and Pathogenesis of the Intracellular Pathogen, Piscirickettsia salmonis |
topic_facet |
Piscirickettsia salmonis agar growth intracellular salmon SRS Aquaculture and Fisheries Bacteriology Cell Biology Environmental Microbiology and Microbial Ecology Immunity Immunology of Infectious Disease Immunopathology Immunoprophylaxis and Therapy Marine Biology Medicinal Chemistry and Pharmaceutics Microbial Physiology Molecular Biology Organismal Biological Physiology Pathogenic Microbiology |
description |
Piscirickettsia salmonis is the causative agent of piscirickettsiosis or salmon rickettsia syndrome (SRS). It is a Gram-negative intracellular bacterium that causes significant mortalities to salmonids, particularly in Chile. The initial isolation and identification of P. salmonis occurred in fish cell lines, therefore growth of P. salmonis is most often carried out in tissue culture. Three cell lines were analyzed in this study to determine which in vitro cellular environment was most suitable for additional research. Tissue culture has been used for the growth and cultivation of P. salmonis for nearly two decades, until the facultative nature of the pathogen was confirmed upon the development of blood and cysteine based agar. Since then, research has continued to drive the creation of novel agar and broth formulations in order to improve the efficacy of cultivation of P. salmonis in cell-free environments. In this thesis, several published agar and broth formulations were tested to understand how this intracellular bacterium grows in these environments. Cell-free media compositions were investigated by using previous formulations and atomic emission spectrophotometry in order to create novel media. This led to the development of MAK broth, MAK2 agar, and BCYE-SRS agar, which significantly improved overall cell-free growth. Furthermore, growth was evaluated using qPCR, SDS-PAGE gels, growth curves, cell counts, colony forming units, and TCID50 titration assays. Additionally, diagnosis of the pathogen was improved by the use of novel cell-free media, vital stains, and PCR assays. In order to study P. salmonis in vivo, two studies were conducted. The first used an intraperitoneal chamber model in Atlantic salmon to observe how the bacterium and host responded and intercommunicated among one another. A lethal dose trial was then carried out in freshwater to determine if P. salmonis grown in cell-free media was a viable option for future challenge trials. Overall, in vitro and in vivo studies provided an ... |
format |
Text |
author |
Makrinos, Daniel |
author_facet |
Makrinos, Daniel |
author_sort |
Makrinos, Daniel |
title |
The Growth and Pathogenesis of the Intracellular Pathogen, Piscirickettsia salmonis |
title_short |
The Growth and Pathogenesis of the Intracellular Pathogen, Piscirickettsia salmonis |
title_full |
The Growth and Pathogenesis of the Intracellular Pathogen, Piscirickettsia salmonis |
title_fullStr |
The Growth and Pathogenesis of the Intracellular Pathogen, Piscirickettsia salmonis |
title_full_unstemmed |
The Growth and Pathogenesis of the Intracellular Pathogen, Piscirickettsia salmonis |
title_sort |
growth and pathogenesis of the intracellular pathogen, piscirickettsia salmonis |
publisher |
DigitalCommons@UMaine |
publishDate |
2016 |
url |
https://digitalcommons.library.umaine.edu/etd/2552 https://digitalcommons.library.umaine.edu/context/etd/article/3589/viewcontent/Daniel_L_Makrinos_Final_Thesis_P._salmonis_1_10_17.pdf |
long_lat |
ENVELOPE(162.381,162.381,56.401,56.401) |
geographic |
Mak |
geographic_facet |
Mak |
genre |
Atlantic salmon |
genre_facet |
Atlantic salmon |
op_source |
Electronic Theses and Dissertations |
op_relation |
https://digitalcommons.library.umaine.edu/etd/2552 https://digitalcommons.library.umaine.edu/context/etd/article/3589/viewcontent/Daniel_L_Makrinos_Final_Thesis_P._salmonis_1_10_17.pdf |
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1768384717922500608 |