Non-native parasites pose a significant threat to aquatic bio-diversity and fishery development. Many factors have facilitated the introduction of non-native parasites into England and Wales. Identifying the effects of these parasites and the importance of any changes to wild fish populations represents a considerable challenge. However, in order for the Environment Agency to identify future disease threats, effectively manage resources and implement practical and sustainable control measures, the risks posed by these parasites to fisheries must be better understood. A structured, risk-assessment process for non-native freshwater parasites is proposed. This approach includes procedures for hazard identification, impact assessment, risk management and communication. A quantitative approach to hazard identification provides an initial prediction of impact at the time of introduction to inform decisionmaking and guide preliminary control measures. This is followed by a more comprehensive framework for impact assessment that promotes understanding of changes at host, population and fishery levels. These are placed into context with the economic and ecological value of native resources. An independent 'non-native parasite review group' has been convened to ensure consistency of policy decisions and clear communication of disease risks to interested bodies. It is hoped that this wil assist the management of future invasions and provide a scientifically robust foundation on which to base proportionate control measures. The Category 2 parasites are defined as "species having a significant disease potential when introduced into waters where they do not already exist, or are non-indigenous species with unknown pathogenicity and distribution". Understanding of the dangers posed by these parasites varies considerably. Ergasilus sieboldi, Anguilicola crassus, Bothriocephalus acheilognathi and Lernaea cyprinacea, are well recognised non-native fish pathogens that pose a considerable threat to fisheries. Conversely, the effects of other introduced parasites remain poorly understood. In many cases, a paucity of published literature at the time of introduction has limited a reliable assessment of impact. Four non-native parasites were considered a priority for further study. These were Paraergasilus longidigitus (Copepoda: Poecilostomatoida), Ergasilus briani (Copepoda: Caryophylldae) Poecilostomatoida ), Atractolytocestus huronensis (Cestoda: and Philometroides sanguinea (Nematoda: Philometridae). Pathological, epidemiological and experimental investigations were undertaken to assess the effects of these parasites at host and population levels. P. longidigitus can cause pronounced pathological changes to the olfactory epithelium of infected fish. Although this damage provides the potential for disruption to spawning through reduced sensitivity to reproductive chemical cues, experimental observations combined with seasonality studies suggest that the parasite is unlikely to disrupt reproduction within infected cyprinid fisheries. Similarly, studies conducted on the caryophylidean cestode A. huronensis provide little evidence to suggest that the parasites poses a threat to car fisheries. Damage within the intesinal tract was characterised by relatively mild mechanical and inflammatory changes. Infections of up 213 parasites had no adverse affect upon the conditon of common carp. Based on these findings and the application of the aformentioned risk model, P. longidigitus and A. huronensis are considered to be of low disease risk to fisheries. The pathology of E. briani and P. sanguinea are described for the first time. Literature suggests that both parasites are pathogenic to juvenile fIsh. E. briani causes a number of pathological changes within the gils of juvenile cyprinids, although these remain localised due to the very strict site specificity of the parasite. Migrations of the nematode P. sanguinea causes damage to the fins and caudal musculature of juvenile crucian car. For both parasites, the extent and severity of pathological changes are inversely proprtional to host size. Further studies at the population level are needed before an assessment of impact to fisheries can be made. The difficulties associated with studying the effects of introduced parasites to juvenile fish populations in the wild are recognised. Recommendations for further study are given.

Fish culture for both food and sport utilization has been greatly increasing throughout the world. The editor of Fish Farming Industries (1973) predicted an increase by 1977 in the United States of 83% for catfish producers, 49% for trout farmers, and 91% for bait dealers. Concomitant with the renewed interest in fish culture has been a corresponding need to further understand fish diseases.

Streptococcus agalactiae is a pathogen of multiple hosts. The bacterium, an aetiological agent of septicaemia and meningo-encephalitis in freshwater and saltwater fish species, is considered a major threat to the aquaculture industry, particularly for tilapia. Cattle and humans are however the main known reservoirs for S. agalactiae. In humans, the bacterium (commonly referred to as Group B Streptococcus or GBS) is a member of the commensal microflora of the intestinal and genito-urinary tracts, but it is also a major cause of neonatal invasive disease and an emerging pathogen in adults. In cattle, S. agalactiae is a well-recognized causative agent of mastitis. Numerous studies focusing on S. agalactiae from human and bovine origins have provided insight into the population structure of the bacterium, as well as the genome content and pathogenic mechanisms through identification of virulence determinants. Concerning S. agalactiae from aquatic origins, scientific information mainly focused on case reporting and/or experimental challenges, with a limited or absence of information in terms of pathogenesis, virulence determinants and genotypes of the strains involved. The objective of this study was to enhance our understanding of the molecular epidemiology, host-adaptation and pathogenicity of S. agalactiae in aquatic species, with particular emphasis on tilapia. Firstly, a collection of 33 piscine, amphibian and sea mammal isolates originating from several countries and continents was assembled, with the aim of exploring the population structure and potential host specificity of aquatic S. agalactiae. Isolates were characterised using pulsed-field gel electrophoresis (PFGE), multi-locus sequence typing (MLST), and a standardised 3-set genotyping system comprising molecular serotypes, surface protein gene profiles and mobile genetic element profiles. Two major subpopulations were identified in fish. The first subpopulation consisted of non-haemolytic isolates that belonged to sequence type (ST) 260 or 261, which are STs that have been reported only from teleosts. These isolates exhibited a low level of genetic diversity by PFGE and clustered with other STs that have been reported only in fish. Another common feature was the absence of all surface protein genes or mobile genetic elements targeted as part of the 3-set genotyping and that are usually found in human or bovine isolates. The second subpopulation consisted of β-haemolytic isolates recovered from fish, frogs and sea mammals, and that exhibited medium to high genetic diversity by PFGE. STs identified among these isolates have previously been identified from strains associated with asymptomatic carriage and invasive disease in humans. The human pathogenic strain ST7 serotype Ia was detected in fish from Asia. Moreover, ST283 serotype III-4 and its novel single locus variant ST491 detected in fish from Southeast Asia shared a 3-set genotype identical to that of an emerging ST283 clone associated with invasive disease of adult humans in Asia. These observations suggested that some strains of aquatic S. agalactiae may present a zoonotic or anthroponotic hazard. STs found among the seal isolates (ST23) have also been reported from humans and numerous other host species, but never from teleosts. This work provided an excellent basis for exploration of the virulence of selected strains in experimental challenges. The virulence of two strains of S. agalactiae was experimentally investigated by intra-peritoneal infection of Nile tilapia (Oreochromis niloticus), using an isolate originally recovered from fish and belonging to ST260, and an isolate originating from a grey seal and belonging to ST23. The clinical signs, the in vivo distribution of viable bacteria and bacterial antigens, and the gross and histopathological lesions that developed during the time course of the infection were investigated. The ST260 strain was highly virulent, whereas no major clinical sign or mortalities occurred in the fish challenged with the ST23 strain. After injection, both strains however gained access to the bloodstream and viable bacteria were recovered from all organs under investigation. During the early stages of infection, bacteria were mostly found within the reticulo-endothelial system of the spleen and kidney. Thereafter, the ST260 demonstrated a particular tropism for the brain and the heart, but granulomatous inflammation and associated necrotic lesions were observed in all organs. ST23 was responsible for a mixed inflammatory response associated with the presence of bacteria in the choroid rete and in the pancreatic tissue only. After 7 days post-challenge and for both strain, the formation or containment of bacteria within granulomata or other encapsulated structures appeared to be a major component of the fish response. However, the load of viable bacteria remained high within organs of fish infected with ST260, suggesting that, unlike ST23, this strain is able to survive within macrophages and/or to evade the immune system of the fish. This work demonstrates that the lack of report of ST23 strains in fish is possibly not due to a lack of exposure but to a lack of virulence in this host. The two strains, which differ in prevalence and virulence in fish, provide an excellent basis to investigate genomic differences underlying the host-association of distinct S. agalactiae subpopulations. The genome of the ST260 strain used in challenge studies was sequenced. We therefore provided the first description for the genome sequence of a non-haemolytic S. agalactiae isolated from tilapia (strain STIR-CD-17) and that belongs by multi-locus sequence typing (MLST) to clonal complex (CC) 552, which corresponds to a presumptive fish-adapted subgroup of S. agalactiae. The genome was compared to 13 S. agalactiae genomes of human (n=7), bovine (n=2), fish (n=3) and unknown (n=1) origins. Phylogenetic analysis based on the core genome identified isolates of CC552 as the most diverged of all S. agalactiae studied. Conversely, genomes from β-haemolytic isolates of CC7 recovered from fish were found to cluster with human isolates of CC7, further supporting the possibility that some strains may represent a zoonotic or anthroponotic hazard. Comparative analysis of the accessory genome enabled the identification of a cluster of genes uniquely shared between CC7 and CC552, which encode proteins that may provide enhanced fitness in specific niches. Other genes identified were specific to STIR-CD-17 or to CC552 based on genomic comparisons; however the extension of this analysis through the PCR screening of a larger population of S. agalactiae suggested that some of these genes may occasionally be present in isolates belonging to CC7. Some of these genes, occurring in clusters, exhibited typical signatures of mobile genetic elements, suggesting their acquisition through horizontal gene transfer. It is not possible to date to determine whether these genes were acquired through intraspecies transfer or through interspecies transfer from the aquatic environment. Finally, general features of STIR-CD-17 highlighted a distinctive genome characterised by an absence of well conserved insertion sequences, an abundance of pseudogenes, a smaller genomic size than normally observed among human or bovine S. agalactiae, and an apparent loss of metabolic functions considered conserved within the bacterial species, indicating that the fish-adapted subgroup of isolates (CC552) has undergone niche restriction. Finally, genes encoding recognised virulence factors in human S. agalactiae were selected and their presence and structural conservation was evaluated within the genome of STIR-CD-17.