O Tomato chlorosis virus (ToCV) é uma espécie do gênero Crinivirus que causa danos, principalmente na cultura do tomateiro (Solanum lycopersicum). Foi primeiramente isolado e descrito em 1998, nos Estados Unidos, e em seguida foi reportado em doze países. No Brasil, foi constatado primeiramente no Estado de São Paulo, na região de Sumaré, em 2008, e posteriormente nos Estados da Bahia, Espírito Santo, Goiás, Minas Gerais e Rio de Janeiro. Há evidência da sua presença também nos Estados do Paraná e Santa Catarina. O ToCV pode infectar outras solanáceas além do tomateiro e, recentemente, foi observado infectando plantas de batata (Solanum tuberosum) no Brasil. Esse crinivirus é transmitido no Brasil principalmente pelo aleirodídeo (mosca branca) Bemisia tabaci MEAM1. Considerando o patossistema batateira/ToCV, não há estudos sobre a ocorrência, sintomatologia em diferentes variedades e danos provocados por esse crinivirus. Também não há antissoro policlonal para o isolado brasileiro do ToCV para uso na diagnose da doença em solanáceas. Esse trabalho teve por objetivos: purificar o ToCV e produzir antissoro policlonal; avaliar a reação de genótipos de batateira à infecção com o ToCV; avaliar o dano provocado por esse vírus em duas variedades de batateira. A purificação do vírus a partir de folhas de tomateiro e a produção de antissoro policlonal em coelho foram satisfatórias. No entanto, o antissoro não foi eficiente em ELISA, mas sim em dot-blot e somente na diluição de 1:20. Foi avaliada a reação de 21 genótipos de batateira à infecção com o ToCV, por meio da inoculação com B. tabaci MEAM1, com chance de escolha do vetor. Nenhum genótipo exibiu resistência à infecção; enquanto a variedade Camila foi assintomática e não apresentou alteração na fotossíntese. Plantas de batateira das variedades Ágata e Asterix sadias foram inoculadas com o ToCV, por meio da B. tabaci MEAM1 e ao final foram avaliadas a massa fresca da parte aérea, peso e número dos tubérculos colhidos. Em dois experimentos independentes, as reduções médias no peso fresco da parte aérea foram de 60,1% para Ágata e 46% para Asterix. Porém, as reduções nas produções dessas variedades, no primeiro experimento foram de 99,5% e 98,1%, respectivamente; enquanto no segundo os valores foram de 82,3% e 56,2%, respectivamente. / Tomato chlorosis virus (ToCV) is a species of the genus Crinivirus, which is causing considerable losses mainly on tomato crop (Solanum lycopersicum). It was first isolated and described on 1998 in the United States and subsequently reported in twelve countries. In Brazil, it was first reported in São Paulo State, in Sumaré region in 2008, and after that on the states of Bahia, Espírito Santo, Goiás, Minas Gerais and Rio de Janeiro. There is evidence of the presence of ToCV on the states of Paraná and Santa Catarina. ToCV can also infect other solanaceae and more recently, it was reported infecting potato plants (Solanum tuberosum) in Brazil. This crinivirus is transmitted by Bemisia tabaci MEAM1. Considering the patosystem potato/ToCV, there are no studies on the occurrence, symptomatology in different varieties, and damages caused by this crinivirus. In addition, there is no polyclonal antiserum for the Brazilian isolate of ToCV for use in diagnosis. The objectives of the present work were: to purify the virus and produce a polyclonal antiserum; to evaluate the reaction of potato genotypes to ToCV infection; to evaluate the yield loss caused by this crinivirus on two potato cultivars. The virus purification from tomato leaves and the production of polyclonal antiserum in rabbit were satisfactorily accomplished. However, the antiserum was not efficient on ELISA test, but in dot-blot, only when diluted 1:20. The reaction of 21 potato genotypes to infection with ToCV was evaluated by inoculation with B. tabaci MEAM1, with chance of choice for the vector. All genotypes were infected with ToCV and Camila was the only one asymptomatic. Plants of cultivars Ágata and Asterix were inoculated with ToCV, by means of viruliferous vector, and at the end were evaluated for the fresh mass of the aerial part, weight and number of harvested tubers. In two independent experiments, average reductions in aerial fresh weight were 60.1% for Ágata and 46% for Asterix. However, reductions in yield of these varieties in the first experiment were 99.5% and 98.1%, respectively; while in the second the values were 82.3% and 56.2%, respectively.

O Tomato chlorosis virus (ToCV) é uma espécie do gênero Crinivirus que causa danos, principalmente na cultura do tomateiro (Solanum lycopersicum). Foi primeiramente isolado e descrito em 1998, nos Estados Unidos, e em seguida foi reportado em doze países. No Brasil, foi constatado primeiramente no Estado de São Paulo, na região de Sumaré, em 2008, e posteriormente nos Estados da Bahia, Espírito Santo, Goiás, Minas Gerais e Rio de Janeiro. Há evidência da sua presença também nos Estados do Paraná e Santa Catarina. O ToCV pode infectar outras solanáceas além do tomateiro e, recentemente, foi observado infectando plantas de batata (Solanum tuberosum) no Brasil. Esse crinivirus é transmitido no Brasil principalmente pelo aleirodídeo (mosca branca) Bemisia tabaci MEAM1. Considerando o patossistema batateira/ToCV, não há estudos sobre a ocorrência, sintomatologia em diferentes variedades e danos provocados por esse crinivirus. Também não há antissoro policlonal para o isolado brasileiro do ToCV para uso na diagnose da doença em solanáceas. Esse trabalho teve por objetivos: purificar o ToCV e produzir antissoro policlonal; avaliar a reação de genótipos de batateira à infecção com o ToCV; avaliar o dano provocado por esse vírus em duas variedades de batateira. A purificação do vírus a partir de folhas de tomateiro e a produção de antissoro policlonal em coelho foram satisfatórias. No entanto, o antissoro não foi eficiente em ELISA, mas sim em dot-blot e somente na diluição de 1:20. Foi avaliada a reação de 21 genótipos de batateira à infecção com o ToCV, por meio da inoculação com B. tabaci MEAM1, com chance de escolha do vetor. Nenhum genótipo exibiu resistência à infecção; enquanto a variedade Camila foi assintomática e não apresentou alteração na fotossíntese. Plantas de batateira das variedades Ágata e Asterix sadias foram inoculadas com o ToCV, por meio da B. tabaci MEAM1 e ao final foram avaliadas a massa fresca da parte aérea, peso e número dos tubérculos colhidos. Em dois experimentos independentes, as reduções médias no peso fresco da parte aérea foram de 60,1% para Ágata e 46% para Asterix. Porém, as reduções nas produções dessas variedades, no primeiro experimento foram de 99,5% e 98,1%, respectivamente; enquanto no segundo os valores foram de 82,3% e 56,2%, respectivamente. / Tomato chlorosis virus (ToCV) is a species of the genus Crinivirus, which is causing considerable losses mainly on tomato crop (Solanum lycopersicum). It was first isolated and described on 1998 in the United States and subsequently reported in twelve countries. In Brazil, it was first reported in São Paulo State, in Sumaré region in 2008, and after that on the states of Bahia, Espírito Santo, Goiás, Minas Gerais and Rio de Janeiro. There is evidence of the presence of ToCV on the states of Paraná and Santa Catarina. ToCV can also infect other solanaceae and more recently, it was reported infecting potato plants (Solanum tuberosum) in Brazil. This crinivirus is transmitted by Bemisia tabaci MEAM1. Considering the patosystem potato/ToCV, there are no studies on the occurrence, symptomatology in different varieties, and damages caused by this crinivirus. In addition, there is no polyclonal antiserum for the Brazilian isolate of ToCV for use in diagnosis. The objectives of the present work were: to purify the virus and produce a polyclonal antiserum; to evaluate the reaction of potato genotypes to ToCV infection; to evaluate the yield loss caused by this crinivirus on two potato cultivars. The virus purification from tomato leaves and the production of polyclonal antiserum in rabbit were satisfactorily accomplished. However, the antiserum was not efficient on ELISA test, but in dot-blot, only when diluted 1:20. The reaction of 21 potato genotypes to infection with ToCV was evaluated by inoculation with B. tabaci MEAM1, with chance of choice for the vector. All genotypes were infected with ToCV and Camila was the only one asymptomatic. Plants of cultivars Ágata and Asterix were inoculated with ToCV, by means of viruliferous vector, and at the end were evaluated for the fresh mass of the aerial part, weight and number of harvested tubers. In two independent experiments, average reductions in aerial fresh weight were 60.1% for Ágata and 46% for Asterix. However, reductions in yield of these varieties in the first experiment were 99.5% and 98.1%, respectively; while in the second the values were 82.3% and 56.2%, respectively.

Thesis(M.S.)--Auburn University, 2007. / Abstract. Vita. Includes bibliographic references.

Made available in DSpace on 2014-12-17T15:32:31Z (GMT). No. of bitstreams: 1 RosileneCS.pdf: 376311 bytes, checksum: 8a0459347381c3b197002ddd72f02225 (MD5) Previous issue date: 2005-11-04 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Oral squamous cell carcinoma (OSCC) is the most common malignancy in oral cavity and human papillomavirus (HPV) may have an important role in its development. The aim of this experiment was to investigate the HPV DNA and viral types in 90 cases of OSCC. Moreover, a comparative analysis between the cases of OSSC with and without HPV DNA was performed by using cell cycle markers p21 and pRb in order to detect a possible correlation of these proteins and HPV infection. DNA was extracted from paraffin embedded tissue and amplified by PCR (polymerase chain reaction) with primers PCO3+ e PCO4+ for a fragment of human β-globin gene. After this procedure, PCR for HPV DNA detection was realized using a pair of generic primers GP5+ e GP6+. Immunohistochemical study was performed by streptoavidin-biotin technique and antibodies against p21 and pRb proteins were employed. Eighty-eight cases were positive for human β-globin gene and HPV DNA was found in 26 (29.5%) of then. It could not be detected significant correlation between HPV and age, sex and anatomical sites of the lesion. The most prevalent viral type was HPV 18 (80.8%). Regarding the immunohistochemical analysis, it was detected significant association between HPV presence and pRb immunoexpression (p=0,044), nevertheless, the same was not observed in relation to p21 protein (p =0,416). It can be concluded that the low detection of HPV DNA in OSCC by the present experiment suggests a possible role of the virus in the development and progression in just a subset of this disease / O carcinoma epiderm?ide oral ? a neoplasia maligna mais freq?ente da cavidade oral e o papilomav?rus humano (HPV) parece ter um relevante papel na indu??o desta les?o. Neste trabalho investigou-se o DNA do HPV e tipos virais em 90 casos de carcinoma epiderm?ide oral (CEO). Realizou-se tamb?m uma an?lise comparativa entre os grupos de CEO com DNA do HPV e sem o DNA do v?rus, empregando-se os marcadores do ciclo celular p21 e pRb, a fim de estabelecer poss?vel correla??o entre a express?o imuno-histoqu?mica dessas prote?nas e a infec??o pelo HPV. O DNA foi extra?do de tecido emblocado em parafina e amplificado por PCR (rea??o em cadeia da polimerase) com um par de primers designados PCO3+ e PCO4+ para um fragmento do gene da β-globina humana. Posteriormente, realizou-se PCR para detec??o do DNA de HPV utilizando-se um par de primers gen?ricos designados GP5+ e GP6+. A tipagem viral foi realizada pela hibridiza??o dot blot. No m?todo imuno-histoqu?mico utilizou-se a t?cnica da streptavidina-biotina com um painel de anticorpos monoclonais para as prote?nas p21 e pRb. Dos 88 casos positivos para o gene da β-globina humana, em 26 (29,5%) foi detectado o DNA do HPV. N?o houve associa??o significativa entre o HPV e as vari?veis idade e sexo dos pacientes e localiza??o anat?mica da les?o. O tipo viral prevalente foi o HPV 18 (80,8%). Quanto ? an?lise imuno-histoqu?mica, foi observada associa??o estatisticamente significativa entre a presen?a do HPV e a express?o imunohistoqu?mica de pRb (p=0,044), entretanto, n?o houve qualquer diferen?a estatisticamente significativa entre a express?o da prote?na p21 e a presen?a do v?rus (p =0,416). P?de-se concluir que o baixo percentual de detec??o do DNA do HPV no carcinoma epiderm?ide oral no presente trabalho, sugere uma poss?vel participa??o do HPV no desenvolvimento e progress?o de apenas um subgrupo dessas les?es

Food poisoning is mainly caused by pathogenic microorganisms and is now a severe problem worldwide. Therefore, rapid and sensitive methods are required to detect foodborne pathogens. A locally isolated bacterium, Bacillus licheniformis B38 was selected for this study. The spores of B. licheniformis B38 were induced by Schaeffer’s sporulation medium containing KCl, MgSO4.7H2O, Ca(NO3)4, MnCl2 and FeSO4. Schaeffer-Fulton endospore staining was used to differentiate spores and vegetative cells, where spores were stained green and vegetative cells were stained red. In order to separate the spores from the cells, a two-phase system was used to obtain pure spore suspension for following experiments. Spore coat proteins were extracted by SDS-8 M urea sample buffer and visualized by two different types of coomassie brilliant blue staining solutions. One of the staining solutions was more suitable for gel elution by diffusion. An ~10 kDa spore coat protein was selected for protein purification. Based on the given results, the protein purification by liquid chromatography was less convincing than using gel elution by diffusion technique. The two hypothetical protein sequences, P06552 and P45693, from the ~10 kDa spore coat protein were identified. In the preliminary study of B. licheniformis B38 spores detection by surface plasmon resonance, several binding parameters were studied. Dot blot was done to verify the reaction between the Bacillus spores polyclonal antibody against the B. licheniformis B38 spore coat protein. The most promising result was the binding of 0.1 mg/mL polyclonal antibody (analyte) to the 0.2 mg/mL spore coat protein at pH 2 (ligand) which showed 5.74 RU. The differences between a dot blot and a SPR detection techniques are described.

Submitted by Hiolanda Rêgo (hiolandarego@gmail.com) on 2016-12-22T15:43:12Z No. of bitstreams: 1 Tese_ICS_Lia Muniz Barretto Fernandes.pdf: 1185581 bytes, checksum: a574788d65eeda6abbf9b9ce1dcc5ee9 (MD5) / Made available in DSpace on 2016-12-22T15:43:12Z (GMT). No. of bitstreams: 1 Tese_ICS_Lia Muniz Barretto Fernandes.pdf: 1185581 bytes, checksum: a574788d65eeda6abbf9b9ce1dcc5ee9 (MD5) / Doença de Newcastle é uma enfermidade viral aguda, altamente contagiosa, que acomete aves de várias espécies, considerada como uma das doenças mais importantes para a indústria avícola moderna. Ferramentas para diagnóstico e controle estão disponíveis para galinhas, porém ainda não foram desenvolvidos testes específicos para avestruzes. O presente trabalho visou padronizar testes sorológicos para a detecção de anticorpos contra a Doença de Newcastle em avestruzes e avaliar a situação soroepidemiológica de plantéis do estado da Bahia e de São Paulo. A padronização da técnica da Inibição da Hemaglutinação revelou interferência do tipo de eritrócito utilizado e demonstrou a necessidade do uso de hemácias da mesma espécie ou, alternativamente, de perus. Testes de ELISA indiretos foram desenvolvidos ou modificados para a utilização nesta espécie e, apesar de apresentarem alta correlação entre si, demonstraram baixa correlação com a HI. Foram desenvolvidos ainda os testes “western blot” e “dotblot”, que podem auxiliar na avaliação da resposta imune e facilitar a implantação de programas de controle. As amostras séricas analisadas revelaram a presença de anticorpos e, a ausência de vacinação dos animais avaliados, reforça a hipótese de que as avestruzes estão em contato com o vírus vacinal ou vírus de campo.

Submitted by Marco Antônio de Ramos Chagas (mchagas@ufv.br) on 2016-06-27T15:06:48Z No. of bitstreams: 1 texto completo.pdf: 717860 bytes, checksum: d71c1053a82b292b3ab10f06c3e68ce6 (MD5) / Made available in DSpace on 2016-06-27T15:06:48Z (GMT). No. of bitstreams: 1 texto completo.pdf: 717860 bytes, checksum: d71c1053a82b292b3ab10f06c3e68ce6 (MD5) Previous issue date: 2004-04-20 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A avaliação de viabilidade de uso da técnica de immuno-dot blot para a detecção do vírus da dengue em homogenatos de larvas e de mosquitos Aedes aegypti e larvas de Aedes albopictus foi realizada utilizando-se soros humanos reativos para dengue e soros policlonais antidengue, produzidos em coelhos da raça Nova Zelândia imunizados com suspensões de vírus DEN-1 (Havaí) e DEN-2 (Nova Guiné) purificados e emulsificadas em adjuvante incompleto de Freund. Os resultados demonstraram que a técnica apresenta sensibilidade adequada para a detecção do vírus da dengue em homogenatos de larvas e de mosquitos A. aegypti e larvas de A. albopictus ainda que os títulos dos anti- soros sejam baixos. A presença do vírus foi detectada em larvas do mosquito A. albopictus, o que pode contribuir para o monitoramento do vírus mesmo na ausência de surtos da doença. Pode, inclusive, contribuir para estudos epidemiológicos mais amplos desse vírus por fornecer ferramenta para esses estudos. / The immuno-dot blot technique was tested and standardized for detection of dengue viruses in Aedes aegypti and Aedes albopictus larvae and mosquitoes homogenates. The experiments were carried out with dengue-reactive human sera and with anti-dengue polyclonal antisera prepared in New Zealand male rabbits immunized with DEN-1 (Hawai) and DEN-2 (New Guinea) purified virus suspensions emulsified in Freund incomplete adjuvant. Results showed that the virus can be easily detected by immuno-dot blot in A. aegypti and A. albopictus larvae and mosquitoes homogenates and suggest that this technique, for its simple procedure and easy reading, could be useful for field virus detection. Although it was recently established that these viruses can multiply in A. albopictus larvae, it is not known whether A. albopictus can indeed vector the DEN-1 and DEN-2 viruses in Brazil and, if so, under what percentages, relatively to A. aegypti mosquitoes. This study provides a tool for broader epidemiological studies of these viruses. / Dissertação antiga

Nitrification is the biological process whereby ammonia is converted first to nitrite by ammonia-oxidizing bacteria, and then the nitrite is subsequently converted to nitrate by nitrite-oxidizing bacteria. Ammonia and nitrite levels are closely monitored during treatment of wastewater due to their toxicity to other biological processes. Sybron Chemicals, Inc., is a company that manufactures a nitrifying bioaugmentation culture (1010N) that is used to enhance the naturally occurring levels of biological nitrification. The microbial population of the 1010N product has been examined using a combination of conventional bacteriological methods and modern molecular techniques, with the goal of developing nucleic acid probes that can be used to detect the product in an environmental sample. Small regions of the 16S rRNA genes of the bacteria in 1010N (and two new nitrifying enrichment cultures) were amplified via the polymerase chain reaction (PCR) and analyzed via temperature gradient gel electrophoresis (TGGE). TGGE is a procedure that allows for separation and visualization of individual PCR products that are the same size, based on differences in their sequence. Two of the predominant PCR products in 1010N were purified from the TGGE gel matrix, reamplified via PCR, and sequenced to allow for phylogenetic analysis and nucleic acid probe design. Coincidentally, two strains (NS500-9 and MPN2) that had been isolated from the 1010N mixed consortium and grown in pure culture were found, via TGGE, to have identical 16S rRNA sequences to the PCR products under investigation. Nearly the full-length 16S rRNA genes from these two organisms were PCR amplified, cloned, and sequenced in order to provide a basis for more accurate phylogenetic analysis. The two dominant organisms in the 1010N product, NS500-9 and MPN2, were thereby found to be most closely related to Nitrosomonas and Nitrobacter, respectively, in the existing database. Using the nucleic acid sequences of the cloned DNA, organism-specific DNA probes were designed for both NS500-9 and MPN2. Unfortunately, difficulties were encountered in using the probes to monitor 1010N activity levels via quantitative dot blot hybridizations (rRNA-DNA). Therefore, efforts were redirected to using the TGGE semi-quantitatively with an internal PCR standard (Brüggeman, et al., 2000) to estimate original cell numbers of 1010N within a mixed consortium. This method was not applicable to our system due to substantial preferential binding of the primers to template other than the standard. Samples from a laboratory-scale bioreactor, bioaugmented with 1010N, were used in an attempt to correlate an increase in activity with a detectable shift in population via TGGE. No detectable shift in population was detected in these samples even though the system exhibited increased levels of nitrification. Therefore, the sensitivity of the TGGE system was also examined by determining the limits of detection when 1010N was present in activated sludge. In both whole cell spiking experiments and genomic DNA spiking experiments, it was found that 1010N must be present at a level of at least 5% of the total population in order to be detected. While this provides some information about microbial populations, in order to evaluate the biological activity of a system, nucleic acid probes should be used in a rRNA based study. / Master of Science

Marginale Parodontitis, die multikausale Erkrankung des Parodonts ist eine Infektionskrankheit, modifiziert durch Wirtsfaktoren und äußere Einflüße. Die als pathogene Mischflora bezeichnete Kombination kommensaler Mikroorganismen spielt die primäre Rolle in der Ätiopathogenese der Parodontitis. In der Aufstellung des Studienziels wurden einzelne Bakterienarten (T. forsythensis, P. gingivalis, A. actinomycetemcomitans, C. rectus, F. nucleatum, Fusobacterium spp., P. intermedia, E. corrodens, V. parvula und C. ochracea) ausgewählt, die eventuell als "Markerkeime" in der aggressiven Form der Parodontitis betrachtet werden können. Dazu wurde eine Kontrollgruppe untersucht, die eine gesunde parodontale Flora besitzt. Die angewandte Nachweismethode basiert auf der PCR-Amplifikation von 16S rDNA und darauffolgender dot-blot Hybridisierung mit Oligonukleotidsonden. Die entsprechenden Sonden wurden hergestellt, optimiert und evaluiert. Für die epidemiologische Untersuchung wurde subgingivale Plaque von vier Parodontaltaschen und einer Kontrollstelle von 45 Patienten mit aggressiver Parodontitis, sowie an fünf Stellen von 21 Senioren entnommen. Die Prävalenz der einzelnen Bakterienarten wurde mit Hilfe des Chi-Quadrat Test verglichen. Obgleich eine hohe interindividuelle Variabilität der Kolonisationsmuster zu beobachten war, konnten T. forsythensis, P. gingivalis, C. rectus und F. nucleatum signifikant häufiger in den Parodontaltaschen als an den gesunden Stellen nachgewiesen werden und können deswegen als "Leitkeime" der aggressiven Parodontitis angesehen werden. A. actinomycetemcomitans konnte nur bei einzelnen Patienten mit aggressiver Parodontitis festgestellt werden. Die Ergebnisse für P. intermedia und E. corrodens ließen keine eindeutige Assoziation sowohl mit der aggressiven Parodontitis als auch mit dem gesunden Parodontalzustand zu. Bei Senioren wurde C. ochracea besonders häufig nachgewiesen. Die Ergebnisse dieser Studie bewiesen die erfolgreiche Einsetzbarkeit der hergestellten Oligonukleotidsonden. / A multifactorial risk pattern of periodontitis has been recognized, where in addition to host and environmental factors, a pathogenic microbiota plays a primary role. The purpose of the current research was to analyze the prevalence of periodontitis-associated microorganisms in patients with aggressive periodontitis and periodontally healthy elders by using molecular-biologic detection methods like eubacterial PCR-amplification of 16S rDNA in combination with dot-blot hybridization. The oligonucleotide probes for the detection of T. forsythensis, P. gingivalis, A. actinomycetemcomitans, C. rectus, F. nucleatum, Fusobacterium spp., P. intermedia, E. corrodens, V. parvula and C. ochracea were designed and evaluated. The PCR products of 42 cultivated target and closely related bacteria were used for the optimization of hybridization conditions. For the epidemiological study, subgingival plaque was sampled from four pockets and one healthy site of 45 aggressive periodontitis patients as well as from five sites of 21 elderly. The differences in the prevalence of bacterial species were analyzed by the chi-square test. The data revealed frequent colonization by T. forsythensis, P. gingivalis, F. nucleatum and C. rectus in patients with aggressive periodontitis, however individual variations were obvious. These species could be predominantly identified in periodontal pockets, but were significantly less common in the healthy sites of the periodontitis patients and in the elderly. These putative pathogens can be conclusively determined as the key-bacteria in patients with aggressive periodontitis. No direct association for P. intermedia and E. corrodens with aggressive periodontitis or periodontal health could be seen. A. actinomycetemcomitans could be detected in only a few patients, reducing its suspected importance in the etiology of aggressive periodontitis. C. ochracea was highly prevalent in the well-maintained elderly, suggesting its association with healthy flora. The results of the study confirmed the reliability of the oligonucleotide probes in a specific and sensitive detection of the respective oral species.

A relatively high percentage of porcine <i>Escherichia coli</i> isolates from cases associated with neonatal and post-weaning diarrhea are positive for the gene encoding the adhesin involved in diffuse adherence I (AIDA-I). This gene and its corresponding protein were first identified and characterized in <i>E. coli</i> strain 2787 isolated from human infantile diarrhea. Little is known about the role of the AIDA-I protein in pathogenesis of porcine enteric disease caused by AIDA-I positive E. coli and the properties of AIDA-I protein expressed by porcine AIDA-I positive <i>E. coli</i> isolates and its receptors. <p>In this study, we demonstrated that AIDA-I adhesin isolated from porcine AIDA-I positive <i>E. coli</i> PD20 and PD58 is an acidic protein consisting of five isoforms. It has a molecular weight (100 kDa) similar to the AIDA-I adhesin expressed by human AIDA-I positive <i>E. coli</i> strain 2787 and has a relatively high amino acid homology (78-87%) with it. Immunodetection of AIDA-I positive <i>E. coli</i> strains using polyclonal anti-AIDA-I antibodies had relatively low sensitivity and specificity, accordingly these tests are unlikely to be used for regular diagnostic detection. <p>Using affinity chromatography, we isolated from porcine intestinal mucus proteins that bind to purified AIDA-I adhesin. These proteins were separated by one- and two-dimensional electrophoresis and subjected to overlay Western blot with purified AIDA-I adhesin and AIDA-I positive <i>E. coli</i> to demonstrate 65 and 120 kDa (p65 and p120) proteins as AIDA-I binding proteins. The identity of p65 was not determined based on LCMS/MS data, whereas p120 was matched to two nuclear proteins (namely, DNA damage binding protein and splicing factor 3b) and one cytoplasmic protein, which is an IgG Fc binding protein. Based on similar amino acid homology, molecular weight, structural similarity to mucin and reported evidence of being secreted by goblet cells into the intestinal lumen, we think that the IgG Fc binding protein is the most likely candidate to serve as a potential receptor in intestinal mucus for AIDA-I adhesin.