Visceral toxicosis of catfish (VTC) is a sporadic, often devastating disease in catfish aquaculture, caused by botulinum neurotoxin serotype/E (BoNT/E). The median lethal dose of BoNT/E for channel catfish (Ictalurus punctatus) fingerlings is 13.7 pg/fish. The diagnosis of VTC is suspected if clinical signs and lesions are produced when affected serum is injected into sentinel-catfish and confirmed if this effect is neutralized with BoNT/E antitoxin. However, the assay is complicated in diagnostic cases by small serum samples from affected fish, lack of standardization and availability of small catfish (~10g). Therefore a zebrafish (~0.4g) bioassay for VTC diagnosis was tested and validated. Susceptibility was tested with other serotypes of toxin to help establish zebrafish as a diagnostic model for BoNTs. BoNT/E consist of100kD heavy chain (HC) and 50kD light chain (LC) linked by a disulfide bond. The HC transports the LC into the cytosol of the neuron, where LC (Zn2+-endoprotease) cleaves the SNAP-25 protein thereby blocking the signal transduction at the neuromuscular junction. The HC-based vaccines can induce protective immunity in mammals. To evaluate HC immunogenicity in catfish, rBoNT/E/HC vaccine produced by USAMRIID was tested; this vaccine did not induce a robust antibody response, but western blot analysis demonstrated specific antibody production in 3 of 11 vaccinated fish. We then developed four channel catfish virus (Ictaluridherpesvirus 1, CCV) recombinants expressing synthetic BoNT/E/HC using our established Gateway CCV recombination system to determine if the virus vector could improve the response. Catfish were vaccinated with these recombinants or with a control vector that expressed Escherichia coli beta-galactosidase (CCV-lacZ). No significant protective immunity or BoNT/E antibodies were observed but CCV-lacZ induced a strong antibody response. These results suggest that BoNT/E/HC has low immunogenicity in channel catfish and deviates from the high immunogenicity observed in mammals. To develop a protective vaccine for VTC, it will be necessary to enhance the BoNT/E /HC immunogenicity in channel catfish. Development of recombinant animals which are resistant to BoNT/E was explored as another potential way to prevent VTC. However, the attempts to modify SNAP-25 of zebrafish by genome editing using Transcription Activator-Like Effector Nucleases were unsuccessful.

No description available.

Interest in the distribution of Clostridium botulinum type E was heightened by the sudden outbreak of human botulism from smoked whitefish chubs and canned tuna fish in 1963. The question arose as to how widely the organism is distributed among fish and shellfish in the Northwest and what potential hazard exists for the consumer of fish products. This sporeforming anaerobic orgnism is heat sensitive and had eluded detection in other surveys where heat shock had been used to eliminate non sporeforming contaminants. More recent study using other techniques than heat to facilitate recovery has shown this organism to be widespread, especially in the marine environment. This study was undertaken to find the incidence and distribution of C. botulinum in the marine organisms and environment of the Pacific Northwest and the food products derived therefrom. All species of fish were examined by incubating the gills and viscera individually in tryptone-peptone-glucose medium anaerobically at 28 C for four days and testing the culture filtrate for mouse toxicity by intraperitoneal injection. Toxic filtrates were typed by retesting them in mice protected by specific botulinal antitoxin of type A, B, E, or F. Among salmonid fish the proportions of specimens of each species yielding toxic filtrates were as follows. Sockeye salmon from the Columbia River, 14 of 59 (23.7%); Chinook salmon from the Columbia River, 19 of 106 (18.0%); Chinook salmon from the Pacific Ocean, 1 of 18 (5,6%); Coho salmon from the Columbia River, 10 of 19 (34.4%); Coho salmon from the Pacific Ocean, 13 of 186 (7.0%); Steelhead trout from the Alsea River, 7 of 37 (19.0%). About one-third of the toxic cultural filtrates were successfully typed and proven to contain botulinal toxin. Most of them proved to be type E toxin but 3 were type A, 3 were type B and one, a comparatively new type, type F, was isolated from a Sockeye salmon in the Columbia River. Pure cultures of Clostridium botulinum type E were isolated from 18 specimens and one specimen yielded a pure culture of type F from a sockeye salmon. This was the second time this type had been isolated. In all of the experimental groups the proportion of fish producing toxigenic cultures was significantly higher in those taken in the two rivers than those of the same species taken from the ocean waters. "Bottom fish" represented by Cod, Sole, Grouper and members of the Sebastodes group were also tested in the manner described above. The number of specimens yielding toxic filtrates were 28 of 157 (17.8%). When grouped according to location at which the fish were caught, those near the mouth of the Columbia River produced a greater percentage of toxic filtrates than did those caught off the open shore line. The results were as follows: Bottom fish from Astoria, 23 of 70 (32.8%), Botton fish from Coos Bay, 5 of 87 (5.6%). Sturgeon specimens produced 3 of 24 (12.5%) toxic filtrates. Most of the species contained type E; however, one type A and one type B were found on typing, with about one-third of the toxic filtrates being successfully typed. Environmental swab samples from the "deep sea" fillet processing plants produced 3 of 39 (7.7%) toxic filtrates. None of the 53 samples taken in the salmon processing plants produced toxic filtrates. Shellfish were collected along the ocean beach and in the estuaries. Three to five shellfish were combined into a single specimen and treated as described. All shellfish obtained from the estuaries demonstrated a higher percentage of toxic filtrates than those obtained from the ocean beach. The results were as follows: Razor clams, 11 of 75 (14.6%), Cockle clams, 12 of 15 (80.0%); Softshell clams, 8 of 12 (66.4%); Littleneck clams, 4 of 11(36.2%); Horseneck clams, 1 of 3 (33.3%); Oysters, 6 of 19 (31.6%); Dungeness crabs, 17 of 24 (71%). Only the razor clams were collected exclusively from the ocean beach. Loss of toxicity on holding mixed cultures at -15 C while awaiting typing was a continual problem. This accounts for only one-third of toxic filtrates being successfully typed. Electrophoretic analysis of the total bacterial proteins was carried out on cell sonicates and cell free culture filtrates by first growing cells for four days at 28 C anaerobically. The cells were separated, washed and disrupted with ultrasonic energy. The cell free culture filtrate was concentrated 10 fold by dialysis against polyethylene glycol 4000. Both the toxigenic organisms and the toxic filtrate demonstrated an extra common protein band in the upper third of the electrophoretic pattern not present in the nontoxic spectra. This band might represent the type E toxin. Differences could also be noted in the number of protein bands in the lower third of the patterns in different nontoxigenic strains and also when the toxigenic and nontoxigenic strains were compared. This could suggest an association with a phage in the toxigenic cultures. / Graduation date: 1970

Thesis (Ph. D.)--University of Wisconsin--Madison, 1970. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliography.

M.Tech. / The development of wrinkles is an aspect of ageing that causes people to become dissatisfied with their appearance. The most obvious or noticeable wrinkles on a person's face are those around and between the eyes. Treatment of, for example the wrinkles between the eyes, such as the most prominent frown lines, leads to a visible improvement of appearance ...

Made available in DSpace on 2014-06-12T23:03:58Z (GMT). No. of bitstreams: 2 arquivo8838_1.pdf: 358153 bytes, checksum: 333a903dffa4635451ecd7b51c36546b (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2004 / A porção comestível do tronco de algumas palmeiras, utilizadas para a produção de palmito é um produto de importância para o Brasil como principal produtor e exportador na forma em coonserva, detendo 85% da produção mundial. A região Nordeste tem se destacado como produtora da variedade pupunha (Bactris gasipaes), nativa da Amazônia, que apresenta vantagens ecológicas por se tratar de palmeira cultivada, com características de perfilhamento, precocidade de produção e qualidade satisfatória. A ocorrência de toxinfecções por Clostridium botulinum provocadas pelo consumo de palmito industrializado levou o Ministério da Saúde a estabelecer a obrigatoriedade do uso de ácido associado à salmoura para produção de palmito com o objetivo de baixar o pH a limites inferiores a 4,5, desfavorecendo o desenvolvimento do Clostridium botulinum, assegurando portanto a saúde do consumidor. Normalmente o ácido cítrico é utilizado para atender as exigências, no entanto admite-se que outros ácidos podem ser utilizados com vantagem quanto aos aspectos sensoriais. Este trabalho teve por objetivo avaliar o efeito da acidificação do palmito de pupunha, cultivado em Pernambuco, com diferentes ácidos sobre as características do produto em conserva. Foram testados os ácidos: cítrico, málico, láctico e tartárico na estabilidade do produto embalado em frascos de vidro com capacidade para 600 ml e quanto às características organolépticas. Os resultados obtidos demonstraram a viabilidade da acidificação do palmito de pupunha com esses diferentes ácidos sem prejuíso nas características físicas, químicas, organolépticas e microbiológicas

No description available.

This study deals with the effects of botulinum toxin A treatment on pathological muscular hyperactivity in a group of 53 children. The thesis objective was to evaluat eeffect of botulinum toxin on muscle tone, range of movement and motor performance of the patient. The theoretical part summarizes the knowledge about this theme, characterizes the term spasticity and an impact of botulinum toxin on abnormal muscular hyperactivity. The practical part evaluates the effect of botulinum toxin in two groups of patients. The first group consists of 14 children, who were tested before and at least one month after botulinum toxin application. Clinical assessment involved range of movement measurement, the evaluation of spasticity with Modified Ashworth scale, Global Assesment of Spasticity Scale (GSA), assessment of motor performance with the use of GMFM test and Functional Independence Meassure for children WeeFIM and summarized physical therapeutic methods used in treatment of patients before and after the application. The second group consists of 39 patients who administered the GSA survey forms within a period of one month. The results confirmed positive effect of botulinum toxin treatment on reduction of spasticity, increasing of range of movement and improvement of motor performance of children. Powered by...

Comparatively little work has been done to determine the ecology of Clostridium botulinum type E since its initial isolation in the nineteen-thirties. This spore forming, anaerobic microorganism is relatively heat labile and has been missed in ecological surveys in which heat was used to selectively screen for spore formers. Use of gentler methods has, however, facilitated its demonstration in marine sediments throughout the Northern Hemisphere. The type E organism elaborates a highly potent neurotoxin and has been isolated as the causative agent in recent fatalities involving the consumption of fish products. Until recently the organism was not believed to be present in the United States south of the Canadian border. The purpose of this investigation was to determine if the organism could be demonstrated in shellfish, inland lake fish, and sediment samples throughout the Northwest. Samples of coastal shellfish-- including various species of clams, crabs and oysters, varieties of smoked fish products, species of inland lake fish and inland lake, river and coastal sediment samples were examined for the presence of the type E organism. The organism was found, to some extent, in almost every type of sample tested. Type E toxin was demonstrated in incubated samples of shellfish and smoked fish products collected from eleven sites along the Oregon and Washington coast. The organism was found in shore sediments from the tidewater and freshwater areas of the Columbia, Alsea, and Umpqua rivers but could not be demonstrated in sediments taken from saltwater beaches at the mouths of these rivers. The type E organism was also demonstrated in fish from inland lakes in the Oregon Cascade Mountains, in sediments from the shores of these lakes, and along the shores of a river and three reservoirs in this area. These samples were collected between 95 and 120 linear miles from the coast. The isolation of the type E botulinum organism in fish and shellfish products demonstrates that the organism does present a potential hazard which should be recognized by Northwest processors and distributors of these products. High concentrations of the spores of this organism, such as those found in bottom sediments of some of the inland lakes and reservoirs may serve to contaminate fish and other wildlife. Much of the data collected supports a terrestial distribution of the spores. / Graduation date: 1966

Thesis (M.S.)--Oregon State University, 2010. / Printout. Includes bibliographical references (leaves 96-104). Also available on the World Wide Web.