Bacillary Necrosis of Pangasius (BNP) caused by Edwardsiella ictaluri is considered to be the most serious disease occurring in farmed striped catfish (Pangasianodon hypophthalmus) in Vietnam. This disease has had an increasing impact over the last ten years and has been reported to cause 50-90% mortality of stocks during a single outbreak. Data obtained from natural outbreaks of E. ictaluri in striped catfish showed the role of environmental factors in the establishment and progression of this disease. At present, factors affecting the virulence and transmission of E. ictaluri in striped catfish are poorly understood. The central hypothesis of this thesis focuses on the complex picture of the environmental factors and infectivity of E. ictaluri in striped catfish. In this study, 80 isolates of E. ictaluri recovered from natural clinical disease outbreaks occurring in striped catfish farms between 2002 and 2011 located in 4 distinct geographical areas within Vietnam were characterised using a variety of methods. The biochemical profiles showed that E. ictaluri isolates from striped catfish in Vietnam have similar phenotypic characteristics to other E. ictaluri isolates from other infected fish species. These data showed high levels of phenotypic homogeneity between the E. ictaluri isolates investigated. The status of isolates recovered from natural infections over time and from geographically distinct farms was evaluated using pulsed-field gel electrophoresis (PFGE), plasmid profile identification and antibiotic sensitivity tests. The PFGE results showed 6 main groups with a similarity of 82% and the corresponding genotypes of the prevalent isolates illustrated annual differences. Three plasmid groups were identified distributed among the isolates investigated, in which high molecular weight plasmids of approximately 35 and 140 kb were found in two of the groups. Plasmid profiles of the present study did not show any trend of geographical region or year of isolation. The 140 kb plasmid has been considered as a multi-antibiotic resistance plasmid which confers resistance to tetracycline, trimethoprim and sulphonamides. All Vietnamese isolates showed a high level of resistance to Oxolinic acid, Sulfadimethoxine/Ormetoprim (Romet), Oxytetracycline and Amoxicillin. A reproducible bacterial immersion challenge model was developed and the LD60 estimated prior to performing subsequent experimental challenge studies. Fish were exposed to 107 cfu ml-1 of E. ictaluri by immersion for up to 30 seconds, resulting in a cumulative percentage mortality of 63%. Edwardsiella ictaluri was recovered and identified from all the dead and moribund fish during these experiments and affected fish showed similar clinical signs and pathology to those reported from natural E. ictaluri infections. The present study resulted in a successful experimental immersion challenge model for E. ictaluri infection in healthy striped catfish. Cohabitation challenges were also developed and produced 15-40% mortality, typical clinical signs and pathology, and successful recovery of the challenge organism demonstrating horizontal transmission of E. ictaluri in striped catfish. Experimental studies were then conducted to investigate the association between pH or salinity of water and susceptibility to E. ictaluri infection in striped catfish. The first experiments were performed in in vitro conditions in which E. ictaluri isolates were cultured in a variety of pH and salt concentrations. In vivo experiments were then designed where striped catfish were exposed to 107 cfu ml-1 of E. ictaluri for 30 seconds and then held at 4 different water pHs (5.5, 6.5, 7.5 and 8.5) or NaCl concentrations (0, 0.5, 1 and 1.5%). The results of in vitro experiments showed that a pH value between 5.5 to 6.5 and salt concentration between 0-0.5% were optimal for the growth of E. ictaluri. The in vivo experiments demonstrated that the cumulative mortality of striped catfish in water at pH 5 and pH 6 was significantly higher than that of fish maintained in more alkaline water (p<0.05). By contrast, the cumulative mortality of the striped catfish maintained in 0.5% salt concentration was significantly lower than those kept in 0%, 1% and 1.5% salt concentration (p<0.05). Clinical signs, lesions and histopathological changes in the affected fish were consistent with those reported in natural infections. This study highlighted the use of pH 8.5 and salinity of 0.5% NaCl as a means of decreasing the susceptibility of striped catfish to E. ictaluri. In conclusion, this study used a variety of methods in order to enhance the understanding of the biochemical, biophysical characteristics, plasmid profile and antibiotic resistance as well as the relatedness of E. ictaluri isolates recovered from farmed striped catfish in Vietnam. This study provided two reliable and reproducible bacterial challenge models (immersion and cohabitation) and emphasised the link between pH and salinity with the infectivity and pathogenicity of E. ictaluri in striped catfish.

Made available in DSpace on 2014-07-29T15:32:01Z (GMT). No. of bitstreams: 1 Gitair Moreira.pdf: 3199339 bytes, checksum: 54ae8591c642548d5c83145316112eef (MD5) Previous issue date: 2010-12-17 / The goal of this research was a thorough assessment of the biophysical and radiometric characteristics of the cultivated pastures in the Cerrado biome, based on both field and remote sensing data acquired in August 2009 and February 2010. The study area, located in the State of Goiás, Central Brazil, comprises an area of approximately 77.806 km², between latitudes 14º 00 S and 16º 30 S, and longitudes 48º 30 and 51º 10 W. Based on our data and results, we found a substantial difference between the two periods investigated, particularly concerning the green biomass and % green cover. While the total biomass remains mostly unchanged, the green biomass varies from 1.2 to 3.1 t/ha between August and February. Likewise, the % green cover increases as much as 65%. All these data and results, situated within the context of a larger initiative, certainly may be instrumental in promoting a more effective and sustainable pasture management in the Cerrado biome. / A presente pesquisa teve como objetivo identificar as características biofísicas e radiométricas das pastagens cultivadas do bioma Cerrado através de dados de campo e sensoriamento remoto, obtidos em agosto (período seco) e fevereiro (período chuvoso). A área de estudo localiza-se na região Centro-Oeste do Brasil, com 77.806 km², coordenadas geográficas latitude 14º 00 S a 16º 30 S e longitude 48º 30 e 51º 10 W. Com base em nossos dados e resultados há uma diferença significativa entre os dois períodos analisados, no que diz respeito aos valores de biomassa verde e porcentagem de verde, com exceção dos valores de biomassa total. Enquanto para a biomassa total os valores são 7,3 e 7,4 toneladas por hectare, em agosto e fevereiro, respectivamente, a biomassa verde varia de 1,2 para 3,1 toneladas por hectare entre agosto e fevereiro. No caso dos valores de % de verde, esta variação é ainda muito mais significativa, variando de 6 a 65%, respectivamente. A expectativa é de que estes dados, gerados e analisados no âmbito de um projeto maior de pesquisa, que envolve vários componentes temáticos e uma equipe multidisciplinar, permitam uma ocupação mais efetiva e ambientalmente sustentável das pastagens no bioma Cerrado.