The adoption of the materiality concept in social and environmental reporting assurance: a field study approach

This study investigates the logics or values that shape the social and environmental reporting (SER) and SER assurance (SERA) process. The influence of logics is observed through a study of the conceptualisation and operationalisation of the materiality concept by accounting and non-accounting assurors and their assurance statements. We gathered qualitative data from interviews with both accounting and non-accounting assurors. We analysed the interplay between old and new logics that are shaping materiality as a reporting concept in SER. SER is a rich field in which to study the dynamics of change because it is a voluntary, unregulated, qualitative reporting arena. It has a broad, stakeholder audience, where accounting and non-accounting organisations are in competition. There are three key findings. First, the introduction of a new, stakeholder logic has significantly changed the meaning and role of materiality. Second, a more versatile, performative, social understanding of materiality was portrayed by assurors, with a forward-looking rather than a historic focus. Third, competing logics have encouraged different beliefs about materiality, and practices, to develop. This influenced the way assurors theorised the concept and interpreted outcomes. A patchwork of localised understandings of materiality is developing. Policy implications both in SERA and also in financial audit are explored.

Identification and characterization of antigenic proteins potentially expressed during the infectious process of Paracoccidioides brasiliensis

Paracoccidioides brasiliensis causes paracoccidioidomycosis (PCM), a systemic mycosis presenting clinical manifestations ranging from mild to severe forms. A P. brasiliensis cDNA expression library was produced and screened with pooled sera from PCM patients adsorbed against antigens derived from in vitro-grown P. brasiliensis yeast cells. Sequencing DNA inserts from clones reactive with PCM patients sera indicated 35 open reading frames presenting homology to genes involved in metabolic pathways, transport, among other predicted functions. The complete cDNAs encoding aromatic-L-amino-acid decarboxylase (Pbddc), lumazine synthase (Pbls) and a homologue of the high affinity copper transporter (Pbctr3) were obtained. Recombinant proteins PbDDC and PbLS were obtained; a peptide was synthesized for PbCTR3. The proteins and the synthetic peptide were recognized by sera of patients with confirmed PCM and not by sera of healthy patients. Using the in vivo-induced antigen technology (IVIAT), we identified immunogenic proteins expressed at high levels during infection. Quantitative real time RTPCR demonstrated high transcript levels of Pbddc, Pbls and Pbctr3 in yeast cells infecting macrophages. Transcripts in yeast cells derived from spleen and liver of infected mice were also measured by qRT-PCR. Our results suggest a putative role for the immunogenic proteins in the infectious process of P. brasiliensis. (C) 2009 Elsevier Masson SAS. All rights reserved.

Serosurvey for antibodies against Brucella abortus and Leptospira interrogans in pampas deer from Brazil

A survey for antibodies against Brucella abortus, and Leptospira interrogans was conducted on 17 pampas deer (Ozotocerus bezoarticus) from Pantanal Matogrossense (State of Mato Grosso do Sul, Brazil) and on 24 pampas deer from Parque Nacional de Emas (State of Goias, Brazil). Antibodies against B. abortus were detected by plate ag glutination, rose Bengal, and complement fixation tests, antibodies against Leptospira interrogans were detected by the microscopic agglutination test. All sera were negative for B. abortus antibodies and all deer sera from Parque Nacional de Emas were negative for L. interrogans antibodies. Four (24%) of 17 sera from Pantanal Matogrossense were positive for L. interrogans serovar (n = 2) hardjo, wolffi (n = 1) and mini (n = 1). While these diseases do not appear to be of major importance to the health status of Pampas deer, it appears that deer are reservoir for leptospirosis in one of the study areas.

A double antibody sandwich ELISA for rapid diagnosis of virus infection and to measure the humoral response against infectious bursal disease on clinical material

A double antibody sandwich ELISA (DAS-ELISA) was developed and employed for simultaneous direct detection of infectious bursal disease virus (IBDV) from bursal samples and to measure the humoral response, using the same basic immunoreagents, the purified and non-purified antigen, capture antibody and chicken hyperimmune sera were prepared, and standardized for this purpose, the DAS-ELISA was applied to both 80 bursal suspensions and 224 corresponding serum samples from vaccinated and non-vaccinated commercial hocks, Bursae samples were collected at 2 weeks of age, and submitted to histological examination, virus isolation in specific pathogen-free chickens embryos, and the DAS-ELISA technique, Serum titres obtained in indirect ELISA and serum neutralization test were compared with those in DAS-ELISA, the agreement was 80% between DAS-ELISA, and the conventional techniques, with high sensitivity (87%) and specificity (90%).

Evaluation of an ELISA for detection of antibodies to Babesia bigemina in cattle and it's application in an epidemiological survey in Brazil

Um ensaio de imunoadsorção enzimática (ELISA) baseado em antígeno bruto foi avaliado na detecção de anticorpos contra Babesia bigemina. A sensibilidade e a especificidade do teste foram de 98,0% e 99,0%, respectivamente. Concordando com a alta especificidade do teste, não foram verificadas reações cruzadas com soros de bezerros inoculados três vezes com 10(7) merozoítos de Babesia bovis. Com relação à comparação do ELISA com a imunofluorescência indireta (IFAT) na detecção de anticorpos contra B. bigemina em bezerros experimentalmente infectados com cinco isolados brasileiros geograficamente distintos deste hemoparasito, o IFAT foi capaz de detectar anticorpos um dia antes do ELISA na maioria dos soros dos animais. Houve uma boa concordância entre os resultados encontrados no ELISA e no IFAT com soros de bovinos de região de estabilidade enzoótica (k=0.61). No entanto, não houve concordância entre os testes sorológicos com soros de animais de área de instabilidade enzoótica (k=0.33). O ELISA foi empregado em um inquérito epidemiológico com 1.367 soros de quatro municípios do Pantanal de Mato Grosso do Sul e caracterizou esta região como uma área de estabilidade enzoótica, uma vez que as prevalências variaram de 87,7 a 98,9%. Dessa forma, este ELISA, que apresentou alta sensibilidade, especificidade e desempenho similar ao IFAT, pode ser utilizado no diagnóstico sorológico de B. bigemina.

Identification and characterization of antigens and potential virulence factors in Enterococcus

Enterococci are normal flora in the human intestinal tract, and also one of the leading causes of nosocomial infections, with most of the clinical isolates being Enterococcus faecalis and Enterococcus faecium. Despite extensive studies on the antibiotic resistance, the pathogenicity of enterococci is not well understood, especially for E. faecium. To identify potential virulence factors based on their antigenicity during infection, E. faecium genomic libraries were constructed and screened using sera from patients with E. faecium endocarditis. ^ As one of my projects, total polysaccharides were extracted from E. faecalis OG1RF and from two epa mutants constructed previously, TX5179 and TX5180, and western blots with patient sera showed that an immuno-reactive polysaccharide present in wild type OG1RF was not produced by either of the two epa mutants. The epa mutants were more sensitive to ethanol stress, neutrophil killing and neutrophil phagocytosis than the wild type OG1RF. ^ Expression of virulence factors is commonly regulated by two component systems. A BLAST search was performed to identify potential two component systems in the E. faecalis V583 genome database using PhoP/PhoS as query sequences, and 11 gene pairs were identified, seven of which were disrupted in E. faecalis OGIRF. ^ Finally, an in vitro translocation model was established for enterococci. E. faecalis strain OG1RF and E. faecium strain DO were shown to be able to translocate across a T84 monolayer, while E. coli strain DH5α and E. faecalis strain E1 could not. ^ In conclusion, several E. faecium antigens expressed in infection (whose antibodies present in sera from patients with E. faecium endocarditis) were identified, two of which, SagA and GlyA, were characterized and suggested to be involved in cell wall metabolism. E. faecalis epa gene cluster (involving in polysaccharide biosynthesis and known to be involved in virulence of E. faecalis in mice) was shown to be involved in hindering neutrophil killing. Several two-component systems were identified in E. faecalis and two of which, EtaRS and EtbRS, were involved in E. faecalis virulence in a mouse peritonitis model.^

Dimethylsulfide dehydrogenase from rhodovulum sulfidophilum: EPR spectroscopy and biochemical analysis reveal its place in the DMSO reductase family of molybdenum enzymes

Dimethylsulfide (DMS) dehydrogenase catalyses the oxidation of DMS to dimethylsulfoxide. The purified enzyme has three subunits of Mr = 94, 38 and 32 kDa and has an optical spectrum dominated by a b-type cytochrome. The metal ion and nucleotide analysis revealed 0.5 g-atom Mo, 9.8 g-atom Fe and 1.96 mol GMP per tool of enzyme. Taken together, these data indicate that DMS dehydrogenase contains a bis(MGD)Mo cofactor. A comparison of the Nterminal amino acid sequence of DMS dehydrogenase revealed that the Mo-containing ct-subunit was most closely related to the c~-subunits of nitrate reductase (NarG) and selenate reductase (SerA). Similarly, the [~-subunit of DMS dehydrogenase was most closely related to the [3-subunits of nitrate reductase (NarH) and selenate reductase (SerB). Variable temperature X-band EPR spectra (120-2K) of 'as isolated' DMS dehydrogenase showed resonances arising from multiple redox centres, Mo(V), [3Fe-4S] +, [4Fe-4S] ÷. A pH dependent EPR study of the Mo(V) centre in lH20 and 2H20 reveals the presence of three Mo(V) species in equilibrium, Mo(V)-OH2, Mo(V)-X and Mo(V)-OH. Between pH6 and 8.2 the dominant species is Mo(V)-OH2 and Mo(V)-X is a minor component. X is probably the anion, chloride. Comparison of the rhombicity and anisotropy parameters for the Mo(V) species in DMS dehydrogenase with other Mo(V) centres in metalloproteins showed that it was most similar to the low pH nitrite spectrum of E. coli nitrate reductase (NarGHI). The spin Hamiltonian parameters (2.0158, 1.8870, 1.8620) for the [4Fe-4S] + cluster suggests the presence of histidine (N) coordination to iron in this cluster. It is suggested that this unusual [Fe-S] cluster may be associated with a histidine-cysteine rich sequence at the N-terminus of the ct-subunit of DMS dehydrogenase.

Situação epidemiológica da brucelose bovina no Estado de Minas Gerais

Realizou-se um estudo para caracterizar a situação epidemiológica da brucelose bovina no Estado de Minas Gerais. O Estado foi estratificado em sete circuitos produtores. Em cada circuito foram amostradas aleatoriamente cerca de 300 propriedades e, dentro dessas, foi escolhido, de forma aleatória, um número pré-estabelecido de animais, dos quais foi obtida uma amostra de sangue. No total, foram amostrados 20.643 animais, provenientes de 2.204 propriedades. Em cada propriedade visitada aplicou-se um questionário epidemiológico para verificar o tipo de exploração e as práticas zootécnicas e sanitárias que poderiam estar associadas ao risco de infecção pela doença. O protocolo de testes utilizado foi o da triagem com o teste do antígeno acidificado tamponado e a confirmação dos positivos com o teste do 2-mercaptoetanol. O rebanho foi considerado positivo, se pelo menos um animal foi reagente às duas provas sorológicas. As prevalências de focos e de animais infectados do Estado foram de 6,0% [5,0-7,1%] e 1,1% [0,78-1,4%], respectivamente. Os resultados para os circuitos pecuários da prevalência de focos e de animais foram: circuito 1, 4,7% [2,7-7,7%] e 0,82% [0,06-1,6%]; circuito 2, 7,2% [4,6-10,6%] e 1,2% [0,53-1,8%]; circuito 3, 6,8% [4,3-10,0%] e 1,5% [0,47-2,4%]; circuito 4, 6,5% [4,1-9,8%] e 1,1% [0,39-1,7%]; circuito 5, 3,8% [2,0-6,5%] e 0,40% [0,11-0,69%]; circuito 6, 6,2% [3,8-9,6%] e 0,66% [0,29-1,0%]; circuito 7, 11,0% [7,7-15,0%] e 1,7% [0,92-2,6%], respectivamente. Os fatores de risco (odds ratio, OR) associados à condição de foco foram: compra de reprodutores (OR = 1,66 [1,13-2,44]), ocorrência de aborto nos últimos 12 meses (OR = 1,81 [1,26-2,60]) e presença de cervídeos na propriedade (OR = 1,56 [1,08-2,27]). A vacinação contra brucelose foi identificada como fator protetor (OR = 0,38 [0,19-0,79]). Concluiu-se que o programa obrigatório de vacinação de bezerras, iniciado na década de 1990, está sendo eficaz ao reduzir a prevalência em todo o Estado e em todos os sistemas de produção animal. As autoridades sanitárias devem priorizar o controle da compra de animais para reprodução, que não apresentem garantias sanitárias e incorporar essa medida às ações de educativas.

Situação epidemiológica da brucelose bovina no Estado de Goiás

Realizou-se um estudo para caracterizar a situação epidemiológica da brucelose no Estado de Goiás. O Estado foi estratificado em três circuitos produtores. Em cada circuito foram amostradas aleatoriamente 300 propriedades e, dentro dessas, foi escolhido de forma aleatória um número pré-estabelecido de animais, dos quais foi obtida uma amostra de sangue. No total, foram amostrados 10.744 animais, provenientes de 900 propriedades. Em cada propriedade visitada aplicou-se um questionário epidemiológico para verificar o tipo de exploração e as práticas de criação e sanitárias que poderiam estar associadas ao risco de infecção pela doença. O protocolo de testes utilizado foi o da triagem com o teste do antígeno acidificado tamponado e a confirmação dos positivos com o teste do 2-mercaptoetanol. O rebanho foi considerado positivo quando pelo menos um animal foi reagente às duas provas sorológicas. No estrato 1, a prevalência foi de 7,7% [4,7-10,7%] para propriedades, e de 1,4% [0,99-1,7%] para animais. No estrato 2, foi de 19,5% [15,0-24,0%] para propriedades e de 2,6% [2,0-3,1%] para animais. No estrato 3, foi de 21,4% [16,7-26,1] para propriedades e 4,3% [3,7-5,0%] para animais. A prevalência obtida para o Estado foi de 17,5% [14,9-20,2%] para propriedades e de 3,0% [2,7-3,3%] para animais. Os fatores de risco (odds ratio, OR) associados à condição de foco, segundo a análise multivariada, foram: compra de reprodutores a comerciantes de gado (OR = 2,06 [1,12-3,52]), ocorrência de abortos nos últimos 12 meses (OR = 5,83 [3,86-8,8]) e prática de vacinação contra brucelose (OR = 2,07 [1,38-3,09]). Tanto a ocorrência de aborto quanto a vacinação são, neste caso, consequência da presença de brucelose no rebanho.

Situação epidemiológica da brucelose bovina no Estado do Espírito Santo

Realizou-se um estudo para caracterizar a situação epidemiológica da brucelose bovina no Estado do Espírito Santo. O Estado foi dividido em dois circuitos produtores. Em cada circuito foram amostradas aleatoriamente cerca de 300 propriedades e, dentro dessas, foi escolhido de forma aleatória um número pré-estabelecido de animais, dos quais foi obtida uma amostra de sangue. No total, foram amostrados 5.351 animais, provenientes de 622 propriedades. Em cada propriedade amostrada foi aplicado um questionário epidemiológico para verificar o tipo de exploração e as práticas de criação e sanitárias que poderiam estar associadas ao risco de infecção pela doença. O protocolo de testes utilizado foi o da triagem com o teste do antígeno acidificado tamponado e o reteste dos positivos com o teste do 2-mercaptoetanol. O rebanho foi considerado positivo quando pelo menos um animal foi reagente às duas provas sorológicas. Para o Estado, as prevalências de focos e de animais infectados foram, respectivamente, de 9,0% [7,0-11,6%] e 3,5% [1,9-6,4%]. Para os circuitos, as prevalências de focos e de animais infectados foram, respectivamente, de: circuito 1, 6,8% [4,5-10,2%] e 3,4% [1,3-8,6%]; circuito 2, 10,9% [7,9%-14,8%] e 3,7% [2,1-6,3%]. Os fatores de risco (odds ratio, OR) associados à condição de foco foram: utilização de inseminação artificial (OR = 7,05 [2,51-19,82]) e confinamento/semiconfinamento dos animais (OR = 2,98 [1,22-7,26]). A vacinação de fêmeas entre três e oito meses de idade foi um fator protetor (OR = 0,03 [0,01-0,1]).

Situação epidemiológica da brucelose bovina no Distrito Federal

Realizou-se um estudo para caracterizar a situação epidemiológica da brucelose bovina no Distrito Federal (DF). No total foram amostrados 2.019 animais, provenientes de 278 propriedades. Em cada propriedade visitada aplicou-se um questionário epidemiológico para verificar o tipo de exploração e as práticas de criação e sanitárias que poderiam estar associadas ao risco de infecção pela doença. O protocolo utilizado foi o da triagem com o teste do antígeno acidificado tamponado e a confirmação dos positivos com o teste do 2-mercaptoetanol. O rebanho foi considerado positivo quando pelo menos um animal foi reagente às duas provas sorológicas. A prevalência no DF foi de 2,5% [1,0-5,1%] para propriedades e de 0,16% [0,04-0,28%] para animais. Em razão dos resultados encontrados, que permitem pensar em estratégias de erradicação, recomenda-se que o DF intensifique o diagnóstico de brucelose, tanto na forma de testes sorológicos sistemáticos como pela introdução de mecanismos de detecção rápida em laticínios, em ambos os casos a fim de aumentar o número de propriedades certificadas como livres da doença e melhorar a sensibilidade do sistema de vigilância ativa.

Modulation of mucosal immunity in a murine model of food-induced intestinal inflammation

Background Hypersensitivity or uncontrolled responses against dietary antigens can lead to inflammatory disorders like food allergy and current models reflect a variety of causes but do not reveal the detailed modulation of gut immunity in response to food antigens after breakdown in mucosal tolerance. Objective To develop and characterize a murine model for food-induced intestinal inflammation and to demonstrate the modulation of gut immune response by dietary allergenic antigens. Methods C57BL/6 mice were sensitized with peanut proteins, challenged with peanut seeds and their sera and gut segments were collected for subsequent analyses. Results Sensitization and challenged with peanut seeds led to alterations in gut architecture with inflammatory response characterized by oedema in lamina propria and cell infiltrate composed mainly by eosinophils, mast cells, phagocytes, natural killer and plasma cells, together with low percentage of gamma delta(+) and CD4(+)CD25(+)Foxp3(+) cells in Peyer`s patches. These animals also presented high levels of specific IgE and IgG1 in sera and modulation of mucosal immunity was mediated by increased expression of GATA-3, IL-4, IL-13 and TNF-alpha in contrast to low IFN-gamma in the gut. Conclusion A murine model for food-induced intestinal inflammation was characterized in which modulation of gut immunity occurs by peanut antigens in consequence of T-helper type 2 (Th2) allergic response and failure of regulatory mechanisms necessary for mucosa homeostasis, resembling food allergy. This work shed some light on the understanding of the pathogenesis of gastrointestinal disorders and intolerance in the gut and supports the development of therapies for food-related enteropathies like food allergy, focusing on gut-specific immune response.

AN ELISA SUITABLE FOR THE DETECTION OF RABIES VIRUS ANTIBODIES IN SERUM SAMPLES FROM HUMAN VACCINATED WITH EITHER CELL-CULTURE VACCINE OR SUCKLING-MOUSE-BRAIN VACCINE

An indirect ELISA for determination of post-vaccination rabies antibody was applied. Purified rabies virus was used as antigen to coat plates, and staphylococcal protein A linked with horseradish peroxidase was used for detecting IgG antibody in human sera. Sera from humans, vaccinated with cell-culture vaccine or suckling-mouse-brain vaccine, were examined. ELISA results were compared to those obtained from the virus neutralization test. The mean and standard deviation of OD were determined for 126 negative sera (pre-vaccination) and for 73 sera from vaccinated persons showing antibody titers lower than 0.5 IU/ml. Results were defined as ELISA -positive, -negative or -doubtful. Establishment of a doubtful region reduced the number of sera otherwise classified as positive (false-positive sera). In this way, the sensitivity, specificity and agreement values were respectively 87.5%, 92.4% and 88.5%. No significant differences were observed in these values when the group vaccinated with cell-culture vaccine and the group vaccinated with suckling-mouse-brain vaccine were compared. It was shown that much of the disagreement between the values obtained by neutralization test and ELISA occurred in sera obtained at the beginning of the immunization process, and was probably due to the presence of IgM in the serum samples, detected only by the former test. This ELISA method can be used as a screening test in rabies laboratories regardless of the kind of vaccine used for immunization.

IgG AVIDITY WESTERN BLOT USING Toxoplasma gondii rGRA-7 CLONED FROM NUCLEOTIDES 39-711 FOR SERODIAGNOSIS OF ACUTE TOXOPLASMOSIS

Toxoplasmosis is an important cause of congenital infection. The present study was performed to evaluate the usefulness of recombinant (r) GRA-7 cloned from nucleotides (n) 39-711 in discriminating between acute and chronic toxoplasmosis. First, commercial IgM, IgG and IgG avidity ELISAs were used to determine the serological profile of the sera. Serum samples were from 20 symptomatic patients with acute infection (low IgG avidity, IgM positive), 10 with chronic infection (high IgG avidity, IgM negative) and 10 with indeterminate IgG avidity (IgM positive) which were tested for IgG avidity status with an in-house developed IgG avidity Western blot using the rGRA-7 recombinant antigen. All 20 sera from cases of probable acute infection showed bands which either faded out completely or reduced significantly in intensity after treatment with 8 M urea, whereas the band intensities of the 10 serum samples from chronic cases remained the same. Of the 10 sera with indeterminate IgG avidity status, after treatment with 8 M urea the band intensities with six sera remained the same, two sera had completely faded bands and another two sera had significantly reduced band intensities. Discrimination between acute and chronic toxoplasmosis was successfully performed by the in-house IgG avidity Western blot.

Serological survey for Chagas' disease in school children in the Rio de Janeiro State, Brazil

A serological survey for Chagas' disease was carried out in school children in the Rio de Janeiro State, a zone considered as non-endemic for the infection. A total of 168 schools in 20 municipalities have been visited and 13,254 blood samples were obtained. The blood eluates were screened by the indirect fluorescence test (IFT), and all positive samples were checked and confirmed in sera by the complement fixation test (CFT). AH serologically positive children were subject to a clinical scrutiny, and the houses where the children lived have been searched for triatomine bugs. Only in two municipalities, Magé and Araruama, there was a significant number of children found positive. The total number of reactive samples by IFT and CFT from 13,004 blood samples screened was 143 (1.00 per cent). No serious clinicai symptoms suggestive of Chagas' disease have been found in any of the positive children, and no triatomine bugs were discovered in the dwellings where the children lived. The overall small percentage of children with positive serology postulates that the infection is not a serious health problem in the area investigated. It is recommended, however, to carry out a more detailed study in Magé and Araruama to find the reason for the relatively high percentage of serologically positive children encountered in these two municipalities.