Thesis (Ph. D.)--University of Wisconsin--Madison, 1955. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 67-69).

Memoria para optar al Título Profesional de Médico Veterinario / Este estudio epidemiológico evaluó la efectividad de vacunas a partir de las mortalidades acumuladas atribuibles a vibriosis (Vibrio sp.) en la especie Salmon salar. Además, se comparó la efectividad de las vacunas comerciales durante el periodo 2004 – 2005 y la influencia de periodo de desarrollo de inmunidad (PDI), macrozona de origen y uso de antibióticos en la efectividad de las vacunas. Se analizaron las encuestas provenientes de 15 centros de las regiones X y XI de Chile, pertenecientes a 8 empresas. Los resultados confirmaron que la vacunación efectivamente es un manejo preventivo contra la presencia de Vibriosis (RR = 0,34) y que existen diferencias significativas entre los promedios de mortalidades entre los grupos vacunados y no vacunados. Mediante análisis de varianza se observaron diferencias significativas entre las mortalidades de las vacunas comerciales. Al evaluar PDI, macrozona y uso de antibióticos (Análisis de varianza factorial), éstos resultaron influir en la eficacia de las vacunas comerciales utilizadas por los centros participantes en el estudio. No se pudo evaluar la interacción entre PDI y vacunas. Estos resultados indican que factores, como los antes mencionados, pueden influenciar positiva o negativamente el desempeño de las vacunas

Memoria para optar el título de Bioquímico / La aparición de vibriosis en los cultivos de peces en Chile ha causado masivas mortalidades de salmónidos cultivados en el sur de nuestro país. Debido a la gran trascendencia económica de esta patología se han estudiado 77 aislados, los cuales han sido identificados a nivel de especie y caracterizados genética, molecular y antigénicamente con el fin de desarrollar futuras medidas de prevención. El análisis de la secuencia del gen rRNA 16S de todos los aislados, permitió identificarlos dentro de las especies Listonella anguillarum (antes Vibrio anguillarum) y Vibrio ordalii. Trece aislados se relacionaron genéticamente a la especie L. anguillarum y un grupo mayoritario de 61 aislados presentó homología con la secuencia tipo de V. ordalii. Desde el punto de vista genético, molecular y antigénico, las cepas chilenas de V. ordalii constituyen un grupo homogéneo. Por el contrario, las cepas de L. anguillarum son heterogéneas en cada una de las propiedades señaladas anteriormente. De esta forma, mediante la técnica de electroforésis de campo pulsado (PFGE) fue posible observar al menos 3 grupos genéticos dentro de las cepas chilenas de L. anguillarum. Sin embargo, al emplear métodos basados en la Reacción en Cadena de la Polimerasa de elementos repetitivos (PCR-ERIC y PCRREP) fue posible obtener mayores diferencias, las que podrían ser empleadas para el seguimiento de un aislado en particular con fines de llevar a cabo estudios epidemiológicos. Además, se determinó la existencia de dos grupos serológicos mayoritarios dentro de los aislados chilenos de L. anguillarun, los cuales no pertenecen a los serotipos O1 y O2 que comunmente están asociados a la vibriosis clásica que afecta a especies salmónideas. Los estudios de virulencia demostraron que las cepas de L. anguillarum y V. ordalii son capaces de causar mortalidad en peces de la especie salmón del Atlántico (Salmo salar), reproduciendo fielmente los signos característicos de ambas patologías. Esta tesis constituye el primer reporte de L. anguillarum como agente causal de vibriosis en salmónidos en Chile

Background Vibrio infection (vibriosis) results from consuming contaminated seafood or exposing skin directly to marine waters or raw seafood. The Centers for Disease Control and Prevention (CDC) estimates that 80,000 illnesses occur each year in the United States. Four species, V. parahaemolyticus, V. vulnificus, V. alginolyticus, and V. cholerae (excluding toxigenic O1 and O139), are responsible for most cases. Understanding foodborne and non-foodborne transmission routes is important for describing epidemiological trends and for directing prevention efforts. Methods Demographic, clinical, and epidemiological data for cases reported between 1988 and 2012 were extracted from CDC’s Cholera and Other Vibrio Illness Surveillance System (COVIS). Outcomes and seasonal trends were described by species and transmission route. Results A total of 10,173 domestically acquired, non-toxigenic cases of vibriosis were reported, including 4,224 (41.5%) V. parahaemolyticus cases, 1,998 (19.6%) V. vulnificus cases, 1,267 (12.5%) V. alginolyticus cases, and 963 (9.5%) V. cholerae cases. There were 4,026 hospitalizations and 795 deaths reported. When categorized by transmission route, 5,775 (56.8%) cases were foodborne and 3,317 (32.6%) were non-foodborne.. Most (52.4%) cases occurred during the summer months with peaks in July and August. Only 140 cases were reported from eight states in 1988 compared to 907 cases reported by 42 states in 2012. The overall crude incidence in 2011 was 0.26 cases per 100,000 population. Discussion The number of reported cases of vibriosis has been increasing steadily since 1988. Increased prevention efforts, including safer seafood products and consumer education, are needed.These efforts should focus on specific populations and transmission routes for each of the top four species that cause most vibriosis cases in the United States.

Many bacteria produce signal molecules that serve in a cell-to-cell communication system termed quorum sensing. This signalling system allows a bacterial population to co-ordinately regulate functions according to their cell number in a defined environment. As bacterial growth progresses towards the stationary phase, signalling molecules accumulate in the growth medium and, above a certain threshold level, regulate the expression of genes involved in diverse functions. Most of the functions monitored by quorum sensing are most beneficial when they are performed as a population than by single cells, such as virulence factor production, biofilm formation, conjugation and bioluminescence. Vibrio anguillarum is a bacterial pathogen that causes terminal hemorrhagic septicaemia in marine fish. V. anguillarum possesses multiple quorum sensing circuits similar to the LuxI/LuxR and the V. harveyi-type systems. In this study, a characterisation of the quorum sensing-regulated transcriptional activator VanT was made. VanT belongs to the V. harveyi LuxR family of transcriptional regulators, which play a central role in quorum sensing signalling in Vibrio species. VanT was shown to regulate serine, metalloprotease, pigment, exopolysaccharide (EPS) and biofilm production. VanT repressed an EPS locus that plays a critical role in bacterial colonization of the fish integument and virulence. The V. harveyi-like quorum sensing systems were shown to limit rather than induce vanT expression throughout growth in V. anguillarum. In contrast to homologous proteins in other Vibrio spp., the quorum sensing phosphorelay protein VanU and the response regulator VanO had antagonistic roles in the regulation of vanT expression. Unlike other members of the luxR family, vanT was expressed at low cell density and no significant induction due to quorum sensing regulation was seen. Interestingly, VanT expression was induced by the alternative sigma factor RpoS as the cells entered stationary phase. RpoS was shown to regulate VanT expression post-transcriptionally by promoting vanT mRNA stability. VanT and RpoS were important for bacterial survival under stress conditions, indicating that VanT is likely an essential factor of V. anguillarum stress response.

Submitted by (edna.saturno@ufrpe.br) on 2016-10-13T12:20:42Z No. of bitstreams: 1 Joanna Dourado.pdf: 721470 bytes, checksum: 1ade18b6e4af854fcb0abb1162f7c2bb (MD5) / Made available in DSpace on 2016-10-13T12:20:42Z (GMT). No. of bitstreams: 1 Joanna Dourado.pdf: 721470 bytes, checksum: 1ade18b6e4af854fcb0abb1162f7c2bb (MD5) Previous issue date: 2009-02-13 / Shrimp farming all over the world has been affected in the last years by many diseases, that have contributed to the development indexes decline in the production of farmed marine shrimp. On top of the losses due to diseases, it's important to mention that some of the vibrio species are zoonotic agents, meaning they can also cause diseases in humans. Given the importance of bacteria of the genus Vibrio to shrimp farming and public health, the aim of this study was to evaluate marine shrimp, correlate the clinical findings with the presence of the vibrio bacteria and identify the isolated species in the water of the ponds of cultivated marine shrimp in Pernambuco. The samples were collected from four coastal farms and the laboratory analysis consisted of the research and identification of the vibrio bacteria in the hepatopancreas and in the water of the tanks, and identification of any alteration by examination of fresh samples. Thiosulphate citrate bile salt agar (TCBS) was used as the culture media and the selected colonies were later biochemically tested. Four hundred and eighty shrimps were analyzed, of hose 424 (88.33%) showed to have vibrio and one or more alterations. There were identified a total of 903 alterations, the most frequent of muscle tissue (329; 36.43%) and the least frequent of the hepatopancreas tubules (121; 13.40%). When analizing the growth cycle, it was observed the largest number of alterations (503; 55.7%) in the dry season. Taking into account the weight of the shrimps, it was observed 540 (59.8%) alterations when the average weight was 8g. Out of all the analyzed animals, 314 (65.42%) showed to have vibrio, being 38 samples from the ponds water and 605 shrimp isolated. The most abundant species was V. mediterranei, (21.31%), considered an environmental species non pathogenic to shrimp, and the least abundant species were V. campbellii, V. rotiferanus, V. shiloni, V. splendidus, V. tapetis and V. wodanis (0.16 %). A dependency between the alterations found on the exam and the presence of the vibrio was noticed, showing that the aforementioned exam, combined to microbiological analyzes, is an important tool that should be used when monitoring the farms. It was also noticed a great variety of pathogenic and non pathogenic species of vibrio in the studied samples. Such microorganisms could, eventually, be related to shrimp and human infections, which could represent a risk for shrimp farming and public health respectively. / A carcinicultura mundial tem enfrentado nos últimos anos vários impactos causados por enfermidades, que contribuíram para a queda dos índices de desenvolvimento na produção de camarão marinho cultivado. Além das perdas causadas devido às enfermidades, ressalta-se que algumas espécies de víbrio são agentes zoonóticos, ou seja, podem causar doenças também em humanos. Dada a importância das bactérias do gênero Vibrio para a carcinicultura e saúde pública, objetivou-se avaliar camarões marinhos, associar os achados clínicos com a presença de bactérias do gênero Vibrio e identificar as espécies isoladas da água dos viveiros e de camarões marinhos (Litopenaeus vannamei) cultivados no litoral do estado de Pernambuco. As amostras foram coletadas em quatro carciniculturas do litoral e as análises laboratoriais compreenderam pesquisa e identificação de bactérias do gênero Víbrio no hepatopâncreas e na água dos viveiros, e identificação de alterações através do exame a fresco. Para o isolamento das espécies foi utilizado o meio de cultivo Ágar Tiossulfato Citrato Sais de Bile Sacarose (TCBS) e as colônias selecionadas foram posteriormente submetidas a provas bioquímicas. Foram analisados no total 480 camarões, destes 424 (88,33%) apresentaram um ou mais tipos de alteração. Foi identificado um total de 903 alterações, sendo a mais freqüente a da musculatura (329; 36,43%) e a menos freqüente a alteração dos túbulos do hepatopâncreas (121; 13,40%). Com relação ao ciclo de cultivo, foi observado o maior número de alterações (503; 55,7%) no período de estio. Levando-se em consideração a média de peso dos camarões verificou-se 540 (59,8%) alterações quando os mesmos estavam com peso médio de 8 g. Do total de animais analisados 314 (65,42%) apresentavam vibrio e uma ou mais alteração no exame a fresco. Foi obtido um total de 643 isolados de Vibrio, sendo 38 de amostras de água dos viveiros e 605 isolados de camarão. A espécie mais abundante foi V. mediterranei (21,31%), considerada espécie ambiental e não patogênica para o camarão, e as espécies de menor predominância foram V. campbellii, V. rotiferianus, V. shiloni, V. splendidus, V. tapetis e V. wodanis (0,16%). Foi verificada uma dependência entre as alterações encontradas no exame a fresco e a presença de vibrio, concluindo-se que o referido exame, associado à análise microbiológica é uma importante ferramenta que deve ser utilizada no monitoramento da saúde dos animais. Verificou-se também uma ampla variedade de espécies patogênicas e não patogênicas de víbrio nas amostras estudadas, que podem eventualmente estar relacionados a infecções nos camarões e nos humanos, representando um risco para a carcinicultura e para a saúde pública respectivamente.

En France, les sites ostréicoles de la méditerranée sont confrontés régulièrement à de fortes mortalités de juvéniles de Crassostrea gigas mais également à des efflorescences récurrentes du dinoflagellé producteur de toxines paralysantes (PSTs), Alexandrium catenella. Parmi les pathogènes associés de manière récurrente à ces mortalités, on retrouve des souches de Vibrio appartenant au clade Splendidus. Nous nous intéressons ici aux interactions entre A. catenella et l’huître C. gigas confrontée à des vibrios pathogènes. Dans une première partie, nous avons montré qu'en conditions expérimentales contrôlées, A. catenella augmentait la sensibilité de l’huître C. gigas au pathogène Vibrio tasmaniensis LGP32. In situ, nous avons également constaté la coïncidence entre la mortalité des huîtres en 2014 et la présence des PSTs dans leurs tissus. Dans une seconde partie, nous avons étudié les interactions entre les PSTs produites par A. catenella et les cellules immunitaires de l’huître, les hémocytes. Un résultat important de cette thèse a été de montrer que la saxitoxine, une des toxines produites par A. catenella, se lie à des structures granulaires présentes dans le cytoplasme des hémocytes de C. gigas et induit une mort hémocytaire caspases-dépendante. Cette mort est indépendante de la production d’espèces réactives de l’oxygène. Nous avons également démontré que la toxine majoritaire des cellules d’A. catenella, la gonyautoxine 5, est la plus toxique sur les hémocytes de C. gigas. Parmi les populations hémocytaires touchées, les hyalinocytes sont très sensibles à ce stress toxique. Les hémocytes étant les cellules immunocompétentes de l’huître qui jouent le rôle central dans la défense contre les infections, nous supposons que leur mort cellulaire induite par les PSTs affecte négativement la défense des mollusques bivalves et explique l'augmentation de la susceptibilité des huîtres à l'infection par Vibrio tasmaniensis LGP32 quand elles sont exposées à A. catenella. / In France, oyster sites in the Mediterranean Sea are regularly confronted to high mortalities of Crassostrea gigas juveniles and to recurrent blooms of the dinoflagellate producer of Paralytic Shellfish Toxins (PSTs), Alexandrium catenella. Among the pathogens associated to these mortalities, we found Vibrio strains belonging to Splendidus clade. We here focus on the interactions between A. catenella and the oyster C. gigas challenged with pathogenic vibrios. In the first part of this work, we have shown that, in vivo, A. catenella increased the susceptibility of the oyster C. gigas to the pathogen Vibrio tasmaniensis LGP32. In situ, we also established the coincidence between oyster mortality in 2014 and the presence of PSTs in their tissues. In the second part of this work, we studied the interactions between the PSTs produced by A. catenella and oyster immune cells, the hemocytes. An important result of this thesis was that saxitoxin, a toxin produced by A. catenella, binds to granular structures in the cytoplasm of C. gigas hemocytes and induces their caspase-dependent cell death. This death is independent of the production of reactive oxygen species. We also demonstrated that the major toxin of A. catenella cells, the gonyautoxin 5, is the most toxic on C. gigas hemocytes. Among affected hemocyte populations, the hyalinocytes are very sensitive to this toxic stress. As hemocytes are oyster immunocompetent cells and therefore play the central role in the defense against infections, we can presume that their cell death induced by the PSTs negatively affects the defense of bivalve mollusks and explains the increased susceptibility of oysters to the infection by Vibrio tasmaniensis LGP32 when exposed to A. catenella.

The identification of climate temperature-sensitive pathogens and infectious diseases is essential in addressing health risks resulting from global warming. Such research is especially crucial in regions where climate change may have a more significant impact like Russia. Recent studies have reasoned that the abundance of V. cholerae tox- is environmentally driven. The aim of the degree project is to investigate the spatial-temporal trends and thermo-climatic sensitivity of non-toxigenic V. cholerae abundance in Russia. This study employed spatial epidemiology tools to identify persistent clusters of the V. cholerae tox- isolation and areas for exploring temperature-depended patterns of the vibrio distribution. Correlation analysis was used to identify regions with temperature-driven Vibrio abundance – vibrioses and the V. cholerae tox- prevalence in water samples. GAM was applied to evaluate the relationship between V. cholerae tox- prevalence and the mean summer air temperature. The spatial analysis detected 16 persistent (7-8 year) clusters of V. cholerae tox- across the study period 2005-2021. The highest-risk areas are located between 54.70 and 55.15 latitudes. The persistent clusters should become targeted areas to improve sanitation conditions. A distinct significant thermo-climatic effect on the abundance of V. cholerae tox- in water basins was found in three Russian regions with temperate marine (the Kaliningrad region - Dwb) and sharp continental climatic conditions (the Irkutsk region - Dwb and the Republic of Sakha - Dwd). Finally, our results showed significant relationships between ambient summer temperature and vibriosis caused by V. cholerae tox- in the Rostov and Volgograd regions. Heat waves for both regions facilitated the eruption of reported vibriosis in 2007 and 2010. The study offers valuable outcomes to support simplified empirical evaluations of the potential hazards of vibrio abundance that might be useful locally for public health authorities and globally as a part of the warning system of climate change effects in Russia.

Three series of studies were conducted to quantify the effectiveness of dietary incorporation of b-1,3-glucan (BG) from Schizophyllum commune in enhancing the immunity and resistance of grass prawn Penaeus monodon to vibriosis and white spot syndrome virus (WSSV) infections. In the first series of studies, three experiments were conducted to evaluate the effectiveness of dietary b-1,3-glucan on shrimp growth and resistance to vibriosis. Weight gain, survival and feed efficiency of juvenile shrimp (0.5 ¡Ó 0.1 g) were not significantly different (P>0.05) after being fed the diets containing BG 0, 0.2, 2, 10 g/kg diet for 18 weeks. Subadult shrimp (20.4 ¡Ó 2.1 g) fed the diet containing of BG 2 g/kg diet for 10 days showed a significantly (P<0.001) enhanced resistance against vibriosis. Postlarvae fed with the BG diet (2 g/kg diet) were more resistant (P<0.001) against starvation and V. harveyi challenges than the postlarvae fed non-BG diet. Additive disease resistance was observed when polyphosphorylated L-ascorbic acid (PAA) was used together with BG. In challenge tests with V. damsela, shrimp fed with PAA (0.2 g/kg diet) + BG (2 g/kg diet) diet for 20 days had a survival rate up to 60%. In the second series of studies, two experiments were conducted to evaluate the effectiveness of dietary b-1,3-glucan on wound healing and immunity in spawners. Dietary supplement of BG reduced the chance of infections, but did not help wound healing as did the supplement of PAA. And regardless of indoor or outdoor rearing, the survival rate of brooder (135 ¡Ó 25 g) fed the BG (2 g/kg diet) diet was higher (P<0.001) than that of the non-BG group. Fed the BG brooders showed enhanced haemocyte phagocytic activity, cell adhesion and superoxide anion production then the control group. Third series of studies evaluated the effectiveness against white spot syndrome virus (WSSV). Six days after being challenged with WSSV, 12.2 % of the BG-treated (2 g/kg diet for 15 days) postlarvae (PL15) and 20 % BG-treated (2 g/kg diet for 20 days) juveniles (5.5 ¡Ó 0.5 g) were still alive; while all non-BG-treated shrimp died. In order to quantify the effectiveness of BG to WSSV, juveniles (6.5 ¡Ó 0.4 g) were fed diets containing graded levels of BG. The results showed that shrimp fed the diet containing BG 10 g/kg for 20 days had the highest (P<0.001) survival rate (42 %) among all groups. Shrimp that received diets supplemented with BG at a dosage >2 g/kg recuperated 9 ~ 12 days after WSSV challenge; while the group fed diets with no or 1 g/kg BG suffered from rapid decrease in total haemocyte count, phagocytic activity, phenoloxidase, O2-, superoxide dismutase (SOD) production and subsequent high mortality. The results in this study showed that b-1,3-glucan is effective in enhancing the phagocytic activity, phenoloxidase, O2- and SOD productions and consequently the resistance of postlarval, juvenile, subadult and brooder P. monodon against vibriosis and viral infections. Since prolonged use of BG, even at optimal dietary levels, decreased the immunity of the shrimp, care therefore must be taken to maximize its effectiveness. A cycle of dietary BG supplement of 2 ~ 10 g/kg diet for 20 days with an intermission of 10 days may serve the purposes.

Depuis une quinzaine d’années, des mortalités estivales d’ormeaux européens, Haliotis tuberculata, surviennent sur le littoral breton et normand, et en structures aquacoles. Ces mortalités sont attribuées à l’espèce bactérienne Vibrio harveyi, et se produisent chez des ormeaux sexuellement matures lorsque la température de l’eau dépasse 17°C.Ce travail de thèse visait en une approche multifactorielle de l’étude de cette interaction hôte-parasite, afin de spécifier les conditions intrinsèques aux ormeaux dans le déclenchement de cette vibriose, le cycle infectieux de V. harveyi chez l’ormeau européen et le rôle de la température dans l’accomplissement de ce cycle infectieux, et enfin la réponse physiologique de l’ormeau lors d’une exposition à V. harveyi.Les principaux résultats montrent un différentiel d’expression génomique entre des ormeaux résistants et des ormeaux sensibles au cours d’une exposition à V. harveyi, attestant ainsi l’importance du statut physiologique de l’hôte dans la survie à la vibriose chez l’ormeau européen. Ce constat est supplémenté de la mise en évidence de sensibilité à cette maladie chez des ormeaux sexuellement immatures, habituellement résistants, acclimatés à 19°C et exposés à des conditions contraignantes de type manipulation. Par ailleurs, l’étude de la voie d’entrée et de la dynamique d’infection de V. harveyi chez l’ormeau européen a révélé un tropisme particulier de ce vibrion pathogène vers les tissus branchiaux dès les premières heures de contact, et son invasion dans le système circulatoire dès 24h de contact. L’étude de la réponse hémocytaire des ormeaux et du métabolisme branchial, à l’échelle moléculaire et cellulaire, lors des premières heures de contact, démontre 1/ la genèse d’un stress oxydatif au niveau des branchies d’ormeaux sensibles à la vibriose et 2/ une altération du fonctionnement des hémocytes, ce qui présume de l’une des stratégies majeures de virulence de V. harveyi. / For fifteen years, summer mortalities have been observed in wild and farmed populations of European abalone, Haliotis tuberculata, along the north French coast. These mortalities are attributed to the bacterial species Vibrio harveyi and occur in sexually mature animals, when the seawater temperature exceeds 17°C.A multifactorial approach to the study of this host-parasite interaction was done during this thesis, in order to specify the intrinsic abalone conditions in vibriosis mortalities, the infectious cycle of V. harveyi in European abalone and the role of temperature in the fulfillment of this infectious cycle, and finally the physiological response of abalone, at cellular and molecular level, when exposed to V. harveyi.The main results showed a differential gene expression between resistant and susceptible abalone during exposure to V. harveyi, indicating the importance of the physiological status of the host, in survival to vibriosis. This hypothesis is supplemented by the susceptibility of sexually immature abalone at 19°C to this disease, usually resistant, and exposed to manipulation stressor. Moreover, the study of the portal of entry and the dynamics of infection by V. harveyi in European abalone revealed a particular tropism of this vibrio pathogen for gill tissues, in the earlier hours of contact, and its invasion into the circulatory system from the first 24 hours of contact. The study of the response of abalone hemocyte and gill metabolism, at the cellular and molecular level, in the earlier hours of contact, shows 1/ a genesis of oxidative stress in gills of susceptible abalone, and 2/ an alteration of hemocyte functions, which may constitute one of the major strategies of virulence in V. harveyi.