Thesis (M.S.)--University of Michigan, 1974.

Thesis (M.S.)--University of Michigan, 1974.

Several final sewage effluents from the Denton Disposal Plant were demonstrated to contain Poliovirus types II and III. Pleated encapsulated filters at pH3.5 enhanced the recovery of the Poliovirus at a higher tier in comparison with nitrocellulose filter (Millipore) and glass fiber filter of pore size 0.45u. This thesis explores problems that face us today in our quest to eliminate viral pathogens from the natural and waste water needed for human, domestic, and industrial consumption. Preliminary experiments concern the use of immunofluorescence, and immunodiffusion techniques as a means of poliovirus identification, which invariably suggests that these techniques may be useful as rapid screening procedures of water samples for presence of potentially pathogenic viruses.

<p>ABSTRACT</p><p>Allergy to mustard has been reported for many years, in some cases as severe anaphylactic reactions. Recent studies imply that this allergy is increasing. Three major allergens have been isolated and characterised; Sin a 1 and Sin a 2 in yellow mustard (Sinapis alba), and Bra j 1 in oriental mustard (Brassica juncea). Yellow mustard and black mustard (Brassica nigra) are the most common species in Europe, whereas oriental mustard is more frequent outside Europe. Mustard plants belong to the Brassicaceae/Cruciferae family. Mustard is present as an ingredient in different foods, sauces and spices, often in small amounts. According to the European labelling directives, mustard and products thereof must always be declared. To monitor this regulation, methods need to be developed to detect mustard. Polyclonal antibodies, produced in rabbits, against yellow and black mustard were characterised with immunodiffusion, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions, and immunoblotting. Rocket-immunoelectrophoresis and enzyme-linked immunosorbent assay (ELISA) were developed for the detection and quantification of mustard protein. With indirect competitive ELISA a concentration of 156ng mustard protein per ml food extract was detected, which is more than enough to cover the lowest reported reactive doses.</p>

ABSTRACT Allergy to mustard has been reported for many years, in some cases as severe anaphylactic reactions. Recent studies imply that this allergy is increasing. Three major allergens have been isolated and characterised; Sin a 1 and Sin a 2 in yellow mustard (Sinapis alba), and Bra j 1 in oriental mustard (Brassica juncea). Yellow mustard and black mustard (Brassica nigra) are the most common species in Europe, whereas oriental mustard is more frequent outside Europe. Mustard plants belong to the Brassicaceae/Cruciferae family. Mustard is present as an ingredient in different foods, sauces and spices, often in small amounts. According to the European labelling directives, mustard and products thereof must always be declared. To monitor this regulation, methods need to be developed to detect mustard. Polyclonal antibodies, produced in rabbits, against yellow and black mustard were characterised with immunodiffusion, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions, and immunoblotting. Rocket-immunoelectrophoresis and enzyme-linked immunosorbent assay (ELISA) were developed for the detection and quantification of mustard protein. With indirect competitive ELISA a concentration of 156ng mustard protein per ml food extract was detected, which is more than enough to cover the lowest reported reactive doses.

Made available in DSpace on 2014-06-11T19:25:38Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-01-24Bitstream added on 2014-06-13T19:33:00Z : No. of bitstreams: 1 lombardi_al_me_araca.pdf: 210693 bytes, checksum: 8b1debf6cc18d982119fcb5ec5619147 (MD5) / A Secretaria de Defesa Agropecuária estabeleceu o controle e erradicação de Maedi-Visna no Brasil, que inclui o diagnóstico laboratorial e sacrifício dos animais positivos. Com o objetivo de se determinar a ocorrência de Maedi-Visna em ovinos na região de Araçatuba-SP, foram coletadas 444 amostras de sangue de ovinos com idade entre dois e 12 anos, de ambos os sexos e de diversas raças, em 20 propriedades desta região escolhidas aleatoriamente. Exame físico geral foi feito em todos animais, incluindo-se freqüência cardíaca (FC), freqüência respiratória (FR) e temperatura retal (TR), afim de diagnosticar sinais clínicos compatíveis com a doença. Utilizou-se o kit para diagnóstico de Maedi-Visna (IDGA - Imunodifusão em gel de ágar) em amostras de soro. Doze animais reagiram positivamente ao teste, em cinco rebanhos distintos. Não houve associação significante entre prevalência da doença e a raça acometida, o sexo dos animais, os tipos de criação e nem entre as variáveis FC, FR e TR. A prevalência da doença foi de 2,7% sendo que nenhum animal positivo apresentou sinais clínicos compatível com Maedi Visna. / The Agriculture Council established measures for control and erradication of Maedi-Visna in Brazil, by the diagnosis and sacrifice of positive animals. The goal of this study was to determine the occurrence of Maedi-Visna in sheep in Araçatuba region – SP. Blood samples were collected from 444 sheep, age range of 2-12 years old, of both sexes and of various breeds, in 20 farms, aleatorily chosen, in this region. Physical examination was performed in all animals, including heart rate, (HR) respiratory rate (RR) rate and rectal temperature (RT) to diagnose the clinical signs compatible with the disease. Agar gel immunodiffusion test (AGID) kit was used to diagnose Maedi-Visna (Biovetech, Pernambuco, Brasil) in serum samples. Twelve animals were positive on the test in 5 different herds. There was no significant interatction between disease prevalence and the breed affected, sex, types of breeding, and HR, RR, or RT. The prevalence of the disease was 2.7% and no animal considered positive for Maedi-Visna showed clinical signs compatible with Maedi-Visna.

Orientador: Luiz Cláudio Nogueira Mendes / Banca: Alexandre Secorun Borges / Banca: Roberto Soares de Castro / Resumo: A Secretaria de Defesa Agropecuária estabeleceu o controle e erradicação de Maedi-Visna no Brasil, que inclui o diagnóstico laboratorial e sacrifício dos animais positivos. Com o objetivo de se determinar a ocorrência de Maedi-Visna em ovinos na região de Araçatuba-SP, foram coletadas 444 amostras de sangue de ovinos com idade entre dois e 12 anos, de ambos os sexos e de diversas raças, em 20 propriedades desta região escolhidas aleatoriamente. Exame físico geral foi feito em todos animais, incluindo-se freqüência cardíaca (FC), freqüência respiratória (FR) e temperatura retal (TR), afim de diagnosticar sinais clínicos compatíveis com a doença. Utilizou-se o kit para diagnóstico de Maedi-Visna (IDGA - Imunodifusão em gel de ágar) em amostras de soro. Doze animais reagiram positivamente ao teste, em cinco rebanhos distintos. Não houve associação significante entre prevalência da doença e a raça acometida, o sexo dos animais, os tipos de criação e nem entre as variáveis FC, FR e TR. A prevalência da doença foi de 2,7% sendo que nenhum animal positivo apresentou sinais clínicos compatível com Maedi Visna. / Abstract: The Agriculture Council established measures for control and erradication of Maedi-Visna in Brazil, by the diagnosis and sacrifice of positive animals. The goal of this study was to determine the occurrence of Maedi-Visna in sheep in Araçatuba region - SP. Blood samples were collected from 444 sheep, age range of 2-12 years old, of both sexes and of various breeds, in 20 farms, aleatorily chosen, in this region. Physical examination was performed in all animals, including heart rate, (HR) respiratory rate (RR) rate and rectal temperature (RT) to diagnose the clinical signs compatible with the disease. Agar gel immunodiffusion test (AGID) kit was used to diagnose Maedi-Visna (Biovetech, Pernambuco, Brasil) in serum samples. Twelve animals were positive on the test in 5 different herds. There was no significant interatction between disease prevalence and the breed affected, sex, types of breeding, and HR, RR, or RT. The prevalence of the disease was 2.7% and no animal considered positive for Maedi-Visna showed clinical signs compatible with Maedi-Visna. / Mestre

Immunoelectrophoresis revealed an antigen-antibody response between 4 day metabolic products and 8, 12 and 16 day sera and between 4 day trypanosomal extract and 16 day serum. Metabolic products from trypanosomes incubated at room temperature do not appear to be antigenic. The limitations of immunodiffusion are discussed in reference to the results. It is suggested that some of the antibodies to metabolic products may be of the precipitating type while others are not. Since a faint reaction also occurred between 4 day trypanosomal extract and 16 day serum, it may be concluded that metabolic products contribute to only a portion of the antibody response of the rat and are by no means the exclusive agents. They possibly work in conjunction with other metabolics within or on the surface of the trypanosome.

<p>Food allergy is an IgE-mediated immunological disease, which affects almost 4% of the adult population and up to 6% of children. Proteins from milk, egg, peanuts, soybean, wheat, fish and nuts are the main cause of food allergies. A less common allergen is pea protein. The National Food Administration analyses undeclared pea protein and contaminations of pea protein in foods using rocket immunoelectrophoresis and immunodiffusion. For both methods an antiserum against pea protein is needed. The aim of this study has been to characterize a newly developed rabbit-antiserum against pea protein. It is important to know if the antiserum is specific against peas, the detection as well as the quantification limits before it can be taken into use. The results of the study show that the antiserum was not absolutely specific, since it cross-reacted with chickpeas, fenugreek and lenses. However there is an "in-house" established PCR-method that can distinguish between chickpeas, fenugreek and peas and that method can be used as a complement to the rocket immunoelectrophoresis. The PCR-method cannot be used alone because it is not quantitative. Rocket immunoelectro¬phoresis detects 0,003% pea protein with purified IgG-antibodies from the antiserum.</p>

<p>Nuts of all kinds are common ingredients in food. For nut allergy sufferers the frequent use of nuts cause problems and "hidden" nuts in food products may elicit allergic reaction when such foods are consumed. Methods for detecting and quantifying walnut (and other nuts) with high sensitivity and specificity are therefore very important.</p><p>The objective of this project was to verify the specificity of a rabbit antiserum against walnut with immunodiffusion and to determine the size of the dominant walnut antigens with Western blotting. In addition, a commercial sandwich ELISA for walnut quantification was validated and compared with a qualitative real-time PCR.</p><p>The rabbit antiserum proved to be less specific but after absorption with cross-reacting nuts and seeds it showed high specificity. The ELISA kit reacted, except for walnut, with pecan and slightly with other nuts and seeds tested. The PCR showed an absolute specificity to walnut. As low levels as 2.5mg walnut/kg can be quantified with the ELISA. This is 8 to 100 fold less than with the PCR method. It is therefore concluded that the ELISA kit is more sensitive than the PCR method but the PCR method is more specific than the ELISA kit.</p>