Since 2007, 96 wild Queensland groupers, Epinephelus lanceolatus, were found dead from Karumba (Gulf of Carpentaria) in Northern Territory to Brisbane in southern Queensland, Australia. In twelve cases, Streptoccocus agalactiae (Group B Streptococcus, GBS) was isolated in pure culture from eyes and internal organs. GBS has been also isolated from other marine species (Javelin grunter, Giant catfish and Squaretail mullet) and caused an outbreak in three species of wild stingrays (Eastern shovelnose ray, Mangrove whipray and Estuary ray) in a public aquarium also in Queensland. These findings have raised public and industry concern over threats to human health from potential transmission between fish and humans, over environmental impacts on other aquatic animals, and over the potential threat posed to the growing Australian aquaculture industry in which, to date, there have been no reported cases of this disease in spite of the devastation caused to the industry overseas. Therefore, my thesis set out to determine whether GBS isolated from dead wild fish in Australia is indeed virulent in Queensland grouper and causative of mortality, to try to identify from where these infections in wild fish originated, to determine potential risk to human health and to other aquatic animals, and to begin to identify mechanisms by which these Australian isolates may cause fatal disease in Queensland grouper. This information may be employed in future risk assessment of potential human transfer, in mitigating further outbreaks in wild fish through understanding origin of infection, and in future development of preventative measures such as vaccination for farmed fish.In order to confirm that S. agalactiae was the cause of the mortality in wild fish, experimental challenges were conducted by injection, immersion and through the oral route. To better understand why these strains of GBS are pathogenic in fish, we conducted a series of cellular immune assays using juvenile Queensland grouper head-kidney leucocytes in the presence of different piscine and terrestrial isolates. Whole-genome sequencing analysis of the piscine and terrestrial isolates provided information on potential origin and potential for transfer between species, and also informed us on critical virulence factors that could be used during pathogenesis of GBS. Moreover, genome analysis gave intriguing insight into the evolution and adaption of GBS to fish as a host and to life in the aquatic environment. To determine putative origin of GBS in wild fish in Australia, multilocus sequence typing (MLST) and capsular polysaccharide (CPS) were derived from whole-genome sequences and revealed that all piscine isolates from Australia were serotype Ib and fell into ST-261 lineage first identified in 1986 from Tilapia in Israel. Overseas, ST-261 has only been found in fish and poikilotherm animals, and has never been associated with humans. High similarity and identical point mutations in the capsular operon of the piscine isolates suggests a common source of infection...