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Background The avian infectious bronchitis virus (IBV) remains a significant source of loss in the poultry industry and early diagnosis is required to prevent the disease from spreading. This study examined the combined use of an ELISA and Western blot (WB) to detect antibodies against the nucleocapsid protein (N) of IBV. The coding sequence for N was amplified by RT-PCR and expressed in Escherichia coli. A soluble recombinant N protein (rN) of approximately 50 kDa was obtained. A total of 389 sera were tested against the rN in ELISA and the results were compared with those of the commercial IDEXX IBV Ab test. ELISA-rN achieved a 90.34% sensitivity and 90.16% specificity. WB confirmed all false negative sera in ELISA-rN or IDEXX test as truly positive. The current study indicate that the combined use of rN in ELISA and WB is a powerful tool for the immunodiagnosis of avian infectious bronchitis. Methods Constructed recombinant pAE/n expression vectors were used to transform E. coli BL21(DE3) Star competent cells (Invitrogen). The rN of infectious bronchitis virus was purified by affinity chromatography using HisTrap HP 1 mL columns pre-packed with pre-charged Ni Sepharose in the ÄKTAprime Automated Liquid Chromatography system (GE Healthcare). A total of 389 serum samples from chickens were used to develop and evaluate the ELISA-rN test. To standardize the indirect ELISA development, serum dilutions (1:100, 1:200 and 1:400) and different concentrations of purified rN antigen (50, 100 and 200 ng/well) were tested. Positive and negative sera for IBV were used as controls. The results were compared with those obtained from a commercial kit. Serum samples scored as negative with the commercial kit but as positive with the ELISA-rN were further analysed by Western blot analyses using the rN protein as an antigen. The results of the ELISA-rN were compared to the commercial kit results using receiver-operating characteristics curves, area under the curve, and confidence intervals with the software GraphPad Prism version 6.0 for Windows (GraphPad Software, USA). Results The expected cDNA fragment of approximately 1240 bp was successfully amplified by PCR using primers designed to select for the coding region of the N protein. The rN was expressed as a soluble protein to avoid the refolding steps and, after purification a yield of 10 mg/L of rN was obtained. The SDS-PAGE results demonstrated the presence of two distinct bands that had a molecular mass of approximately 45 and 50 KDa. Out of 244 sera that scored positive in the commercial ELISA IDEXX IBV Ab Test, 220 were also positive in the ELISA-rN, yielding an ELISA-rN test sensitivity of 90.16%. Out of 145 sera that scored negative in the IDEXX IBV Ab Test, 131 also scored negative in the ELISA-rN, indicating a specificity of 90.34%. Sera that tested negative in the ELISA-rN and positive in the commercial test also reacted with the rN protein in Western blot. Conclusions The association between the ELISA and Western blot techniques developed in this study with a subunit of IBV (rN) were able to detect antibodies that the commercial ELISA did not detect suggesting that the ELISA-rN has greater sensitivity.

Development cooperation (DC) is shaped by norms. We aim at filling a gap of research on DC by using the academic debates in international relations on norms. Contrary to interpretations that consider developed countries as norm-makers and developing countries as norm-takers, our analysis provides evidence that—and highlights how—Southern agents have influenced the processes of norm-setting and norm-diffusion for DC. The OECD was the dominant norms “entrepreneur” for a long period of time; more recently, developing countries have played a significant role in setting DC norms. We identify the diverging norms for official development assistance and South-South cooperation and the interrelationship between both norm systems. Thus, norm-making, norm-taking, and norm-diffusion of two competing norm clusters are key terms of the contribution.

A vacinação é a forma mais utilizada para prevenir a bronquite infecciosa causada pelo vírus da bronquite infecciosa das galinhas (IBV). Contudo, as vacinas convencionais são incapazes de diferenciar aves infectadas de vacinadas. No presente trabalho foi construído, caracterizado, e avaliado como candidato vacinal, um adenovírus recombinante expressando o gene N do IBV. O gene N foi clonado em um adenovírus humano tipo 5 defectivo e transfectado para as células HEK-293A para gerar rAd5_N. Após o vetor ser obtido como esperado e a confirmação da expressão da proteína N em HEK-293ª, foi realizada inoculação pela via oculo-nasal na dose de 107 TCID50/0,1mL para imunização de galinhas livres de patógenos específicos (SPF). A resposta imunológica do Ad5_N e a proteção contra o desafio ao IBV foram avaliadas e comparadas com uma vacina viva comercial. Não foram detectados anticorpos anti-IBV em aves vacinadas com o Ad5_N. A vacina comercial induziu anticorpos detectáveis a partir do 7º dia pós-vacinal. Em aves vacinadas com o Ad5_N não houve aumento na expressão de IFNγ. Neste estudo, o rAd5_N obtido não conferiu proteção contra desafio com IBV-M41. Os resultados indicam a necessidade de avaliar adenovírus recombinantes expressando outros genes do IBV Made available in DSpace on 2019-11-13T18:16:37Z (GMT). No. of bitstreams: 1 Ritterbusch2019.pdf: 730975 bytes, checksum: 0acd3b65f8f3795195d9192f02da5e57 (MD5) Previous issue date: 2019

This article questions why and how the private sector was articulated as a legitimate agent in a field almost entirely dominated, until the 2000s, by DAC donors. We argue that private agents were admitted in the field across the fractures produced by SSCP and throughout a series of normative and managerial adjustments, which we called here regulated improvisations.

During the past two decades the Brazilian Foreign Policy Community (BFPC) experienced a great deal of change. Brazil’s international stance has for a very long time been recognized for its attachment to the principles of sovereignty, non-intervention and non-interference, as well as for its quest for autonomy and status recognition. Nevertheless, since the beginning of the 2000s, Brazil led the peacekeeping operation in Haiti (under Chap. VII of the UN charter), got involved in the military coup in Honduras and engaged itself in contentious development projects in Africa. What has changed? How has the Brazilian foreign policy community bared new understandings about these long-standing principles. This chapter discusses Brazil’s international stance in the field of international security, focusing on the intersection between security and development and the regional-global nexus. We contend that in order to understand the process of change and continuity it is necessary to look into the relations between the positioning of the country, the understandings about the dynamics of the international system and forms of resistance and adaptation considered by those who are making decisions and building discourses (the foreign policy community).

This chapter has as a point of departure the enquiry into the drivers for Brazil’s engagement with South-South cooperation (SSC). Nonetheless, instead of taking SSC as a tool always already available for the foreign policy community, it enquires (i) into the process of manufacturing SSC as a foreign policy tool and, (ii) on its role in shaping the moral claims that Brazilian policy networks brought to the international arena while seeking a new status. The analysis is mainly on the “golden period” of Brazilian SSC from 2003 to 2014. During this decade, the Brazilian foreign policy community strove to change the perception the international community kept about Brazil. This process entailed establishing moral authority in some specific areas of international policymaking, particularly on issues like inequality, poverty, and hunger. During that period, Brazil started to be recognized as a kind of social policies powerhouse. Through a systematic bottom-up analysis, the chapter briefly presents the policy networks related to public health, food and nutrition security and agricultural innovation. These three sectors have played a germane role in shaping Brazil’s moral authority and its international standing as a champion for eradicating hunger, providing access to a universal health system and medicines and feeding the world’s growing population.

Over the past 15 years, the rise of new powers is changing the international agenda, as Brazil, Russia, India, China and South Africa and other emerging powers seek to influence the conduct of multilateralism. The quest for influence is bringing these powers into fields and policy arenas previously reserved for traditional great powers. As a consequence, fields such as trade negotiations, development aid, and international peace and security are undergoing significant changes. These changes raise questions about the role of Brazil in particular. Brazil has adopted a role of leader for the Global South in trade negotiations, made the case for less conditionality and interference in what it sees as sovereign affairs, and involved itself significantly in changing the international peace and security agenda. In all these fields, Brazil has brought new ideas and commitments to the table. Yet, the drivers of specific Brazilian foreign policy engagements remain unclear. Specifically, the new policy areas in which emerging powers are engaging and Brazil’s shift from domestic to international engagement, and in this, its relations to its domestic constituencies, as well as to other rising powers and the established great powers, require a more sustained engagement.

The growing number of development stakeholders and initiatives in developing countries has added complexity to international development cooperation (IDC). Recipient countries have witnessed the increasing presence of emerging countries such as the BRICS (Brazil, Russia, India, China, and South Africa), offering South–South cooperation as an alternative model for development. We call the impact of the new practices of South–South cooperation providers on the prevailing IDC structure the ‘BRICS effect’ – an effect that ultimately destabilises established positions and interaction patterns between agents, and even between traditional donors and recipients. Combining the Bourdieusian notion of social fields with international relations (IR) perspectives on the changing geopolitics of international aid, this article discusses how the BRICS effect challenges established principles and practices from the field of IDC, indicating at least three dimensions: (1) new positions beyond the donor vs recipient dyad; (2) new modes of development cooperation; and (3) transformation of institutional architecture and governance mechanisms.

Membros da familia Herpesviridae , subfamilia Alphaherpesvirinae , herpesvirus bovino tipos 1 (BoHV-1) e 5 (BoHV-5) tem sido associados a diferentes condicoes clinicas em bovinos. Assim, diferenciacao entre BoHV-1 e BoHV-5 e uma valiosa informacao objetivando um melhor entendimento da patogenia e epidemiologia destes virus no rebanho bovino. Contudo, metodos para diferenciacao sao escassos devido as reacoes cruzadas originadas da alta homologia genomica e antigenica entre estes virus. O presente estudo concentrou-se na analise da regiao carboxi-terminal da glicoproteina C (nucleotideos 16763 - 17337 em BoVH-1 e 17671 - 18242 em BoHV-5), uma vez que tal regiao apresentou caracteristicas potenciais que permitissem a diferenciacao entre estes virus. Assim, no primeiro capitulo do presente trabalho a regiao carboxiterminal da gC foi utilizada como alvo para o desenvolvimento de uma Reacao em Cadeia da Polimerase (PCR) seguida da Analise da Restricao Enzimatica (REA) do fragmento amplificado, capaz de detectar isolados de BoHV-1 ou BoHV-5, gerando fragmentos com tamanhos de 575 pares de base (pb) frente a BoHV-1 e 572 pb frente a BoHV-5. A diferenciacao entre BoHV-1 e BoHV-5 foi obtida apos a digestao dos amplicons com a enzima de restricao Bgl I. No segundo capitulo, a regiao carboxiterminal de 23 amostras Sul-Americanas de BoHV-1 ou BoHV-5 foram amplificadas e sequenciadas. A analise filogenetica realizada com as sequencias de nucleotideos revelou identidade variando entre 98,7 a 99,8% (BoHV-1/ BoHV-1), 88,3 a 92% (BoHV-1/ BoHV-5) e 96 a 99,7% (BoHV-5/ BoHV-5). Alinhamentos realizados com sequencias de aminoacidos revelaram identidade variando entre 97,5 a 99,5% (BoHV-1/ BoHV-1), 77,5 a 84,4% (BoHV-1/ BoHV-5) e 92,1 a 99,5% (BoHV-5/ BoHV-5). Analises filogeneticas revelaram, ainda, que: i) alguns isolados de BoHV-5 apresentaram um padrao diferente daqueles ate o presente reconhecidos; ii) uma amostra isolada no Uruguai (BoHV-1.2) apresentou um padrao de agrupamento distinto quando comparado com as outra amostras de BoHV-1.2. O terceiro capitulo apresenta o primeiro relato de deteccao de BoHV-5 de semen bovino atraves da analise antigenica, amplificacao diferencial da regiao amino-terminal do gene da gC de BoHV-1 e 5, seguida da caracterizacao por REA do isolado em estudo. Em sintese, atraves da utilizacao da gC foi possivel avaliar a homologia entre as amostras Sul-Americanas estudadas; desenvolver uma PCR/REA diferencial entre estes virus e relatar a deteccao de uma amostra de BoHV-5 isolada a partir de semen contaminado.

Hepatitis E virus (HEV) is highly disseminated among swine herds worldwide. HEV is also a threat to public health, since particularly genotypes 3 and 4 may cause acute hepatitis in human beings. No previous studies were done on the occurrence of HEV in environmental samples in Rio Grande do Sul, Brazil. In the present study, reverse transcriptase-polymerase chain reaction (RT-PCR) was employed to detect the presence of HEV in swine feces and in effluents from slurry lagoons in farms located in the municipality of Teutônia, inside the area of swine husbandry in the state. Pooled fecal samples from the floor of pig barns from 9 wean-to-finish farms and liquid manure samples were collected from the slurry lagoons from 8 of these farms. From the pooled fecal samples, 8/9 were positive for the HEV ORF1 gene by RT-PCR; all the slurry lagoon samples were positive for HEV RNA (100%). The identity of the HEV ORF1 amplicons was confirmed by sequencing belonging to HEV genotype 3, which was previously shown to be circulating in South America. O vírus da hepatite E (HEV) é altamente disseminado entre rebanhos suínos no mundo todo. O HEV é também uma ameaça à saúde pública, já que os genótipos 3 e 4 podem causar hepatite aguda em seres humanos. Não há estudos anteriores sobre a ocorrência de HEV em amostras ambientais no Rio Grande do Sul. No presente estudo, empregou-se transcrição reversa e reação em cadeia da polimerase (RT-PCR) para detectar a presença de HEV em fezes de suínos e efluentes de lagoas de chorume em fazendas localizadas no município de Teutônia, representativo da região de maior produção de suínos no estado. Pools de amostras fecais foram coletadas a partir do chão de galpões de suínos provenientes de 9 propriedades de terminação; outra amostra de esterco líquido foi coletada das lagoas de chorume de 8 dessas fazendas. A partir das amostras fecais reunidas, 8/9 foram positivas para o gene ORF1 de HEV por PCR convencional; todas as amostras de lagoas de chorume foram positivas para RNA de HEV (100%). A identificação dos produtos de amplificação de HEV ORF1 foi confirmada por sequenciamento pertencente ao HEV genótipo 3, o qual foi previamente detectado na América do Sul.

This article discusses the transformation in development architecture, focusing on the role of emerging powers and the growing relevance of South–South cooperation (ssc). Drawing on a conceptual toolkit based on the work of Pierre Bourdieu, it aims to approach ssc as a narrative and to understand the processes of contestation that have turned international development into a battlefield since the end of the 1990s. The article argues that the emergence of ssc has contributed to decentring the field of international development, both in terms of the agents authorised to play and the practices considered legitimate. Within this process the Global Partnership for Effective Development Cooperation, led by the oecd’s Development Assistance Committee, and the United Nations Development Cooperation Forum have become two sites on the battlefield on which the borders of international development are being redrawn.

This study focuses on the detection of chicken anemia virus (CAV) and avian gyrovirus 2 (AGV2) genomes in commercially available poultry vaccines. A duplex quantitative real-time PCR (dqPCR), capable of identifying genomes of both viruses in a single assay, was employed to determine the viral loads of these agents in commercially available vaccines. Thirty five vaccines from eight manufacturers (32 prepared with live and 3 with inactivated microorganisms) were examined. Genomes of CAV were detected as contaminants in 6/32 live vaccines and in 1/3 inactivated vaccines. The CAV genome loads ranged from 6.4 to 173.4 per 50 ng of vaccine DNA (equivalent to 0.07 to 0.69 genome copies per dose of vaccine). Likewise, AGV2 genomes were detected in 9/32 live vaccines, with viral loads ranging from 93 to 156,187 per 50 ng of vaccine DNA (equivalent to 0.28-9176 genome copies per dose of vaccine). These findings provide evidence for the possibility of contamination of poultry vaccines with CAV and AGV2 and they also emphasize the need of searching for these agents in vaccines in order to ensure the absence of such potential contaminants.

The chapter looks at Brazilian health cooperation by focusing on one aspect that is often overlooked in studies about SSC: the sociology of SSC workers. The analysis departs from the position of the “cooperantes” that conduct Brazilian SSC based on their own policy experiences at home. By telling the story of how the BHEC was founded, the chapter explores the country’s progressive internationalization through an engagement in minilateral arrangements, such as the MERCOSUL, the UNASUL and the CPLP, as well as structuring SSC projects in health. The latter are referred to here as the South–South cooperation fold in health. The SSC fold supposedly gathers together agents from diverse countries around a shared understanding of health and healthcare in the aid relationship. However, taking the case of the health partnership with Mozambique and the establishment of an antiretroviral factory, the chapter argues that the common vocabulary is always subject to particular interpretations. These depend on the networks and sociohistorical processes in which the expert communities are embedded—that is, the agents’ structural positions.

Postcolonial perspectives are indebted to Foucault’s writings in many important ways. In fact, since the publication of Edward Said’s major work Orientalism, postcolonial thinkers often invoke Foucault’s notion of the power-knowledge nexus. Drawing on Foucault, they note that the way we frame events necessarily involves strategies of power and techniques of knowledge. Although Foucault had explored in many ways how power, knowledge and subjectivity are mutually related, his work was quite silent about how they connect with the colonial experience. The main purpose of this chapter is to explore the silences to develop a critical understanding of global politics in Foucault’s writings about European international society and its political implications.

Swine effluents must be correctly handled to avoid negative environmental impacts. In this study, the profiles of two swine manure treatment systems were evaluated: a solid–liquid separation step, followed by an anaerobic reactor, and an aerobic step (System 1); and a biodigester followed by serial lagoons (System 2). Both systems were described by the assessment of chemical, bacterial and viral parameters. The results showed that in System 1, there was reduction of chemicals (COD, phosphorus, total Kjeldhal nitrogen – TKN – and NH3), total coliforms and Escherichia coli; however, the same reduction was not observed for Salmonella sp. Viral particles were significantly reduced but not totally eliminated from the effluent. In System 2, there was a reduction of chemicals, bacteria and viruses with no detection of Salmonella sp., circovirus, parvovirus, and torque teno virus in the effluent. The chemical results indicate that the treated effluent can be reused for cleaning swine facilities. However, the microbiological results show a need of additional treatment to achieve a complete inactivation for cases when direct contact with animals is required.

Samples collected from two swine manure treatment systems including: swine manure treatment system and demonstrative unit (SMTS and DU), were analyzed by qPCR to quantify the amount of porcine adenovirus (PAdV) and porcine circovirus (PCV2) present. Positive samples were tested for virus integrity using DNase assay. Fifty-six water samples were collected monthly from March 2009 to May 2010. PAdV genome was found 66% of the samples in the SMTS and in 78% of the samples in the DU system. PCV2 was detected in 96% of samples collected from the SMTS system and in 86% of samples from DU. DNase assay revealed that there were undamaged virus particles of both PAdV and PCV2 in all sampling sites in the SMTS. However, undamaged particles of both viruses were detected in samples from the DU system in the affluent and middle sites, though undamaged PCV2 was absent in the effluent samples.

Animal and human wastewater can potentially contaminate water sources and the treatment of drinking water may not effectively remove all contaminants, especially viruses. The purpose of the present study was to evaluate the viral contamination of water used for human and animal consumption in the city of Concórdia, located in southern Brazil. Porcine circovirus type 2 (PCV2), porcine adenovirus (PAdV), human adenovirus (HAdV) and human norovirus (NoV) were searched for using quantitative polymerase chain reaction (qPCR). HAdV-positive samples were tested for viral infectivity by plaque assay. The qPCR results showed that PAdV, PCV2 and HAdV genetic material were present in all sampling sites. NoV was absent in all samples. The presence of genetic material from PAdV and PCV2 was detected in 30% and 45% of the 36 analyzed samples, respectively, with an average of 102 gc mL–1 for PAdV and 104 gc mL–1 for PCV2. HAdV was present in 100% of the samples, with an average of 104 gc mL–1. However, in plaque assay, only 36% of the samples were positive. As viable particles of HAdV were found in drinking water, these results confirm that swine manure and human sewage impact surface water and groundwater, endangering water quality and indicating a potential risk to public health.

Les révélations autour des programmes secrets de la NSA ont confirmé l’existence d’une surveillance de grande envergure de nos communications par les autorités gouvernementales américaines, qui touche également les pays alliés des États-Unis en Europe et en Amérique latine. Les ramifications transnationales de la surveillance nous invitent à ré-examiner les pratiques contemporaines des affaires internationales. Le débat ne se limite pas aux relations des États-Unis avec le reste du monde, ni à la surveillance et à la vie privée : il est beaucoup plus large. Cet article collectif décrit les spécificités de la cyber-surveillance, y compris les pratiques hybrides des services de renseignement et des compagnies privées de télécommunications. Il analyse ensuite les impacts de ses pratiques sur la sécurité nationale, la diplomatie, les droits de l’homme, la démocratie, la subjectivité et l’obéissance. After Snowden: Rethinking the Impact of SurveillanceCurrent revelations about the secret US-NSA program, PRISM, have confirmed the large-scale mass surveillance of the telecommunication and electronic messages of governments, companies, and citizens, including the United States’ closest allies in Europe and Latin America. The transnational ramifications of surveillance call for a re-evaluation of contemporary world politics’ practices. The debate cannot be limited to the United States versus the rest of the world or to surveillance versus privacy; much more is at stake. This collective article briefly describes the specificities of cyber mass surveillance, including its mix of the practices of intelligence services and those of private companies providing services around the world. It then investigates the impact of these practices on national security, diplomacy, human rights, democracy, subjectivity, and obedience. Techniques de surveillance de masse et portée mondiale de l’Internet : un fossé permanent ? Un ruban de Moebius de la sécurité nationale et de la surveillance transnationale Accès illimité aux données, sécurité nationale et renseignement étranger : la distribution inégale de la suspicion La sécurité nationale et la numérisation de la raison d’État Un champ d’échanges informatisés entre professionnels de l’information sensible et une guilde tentant de l’organiser De multiples sites de résistance Les jeux des États autour du ruban de Moebius Comment ramener le ruban de Moebius vers les lignes souveraines ? Raison d’État et géopolitique adaptées aux nécessités de l’ère du numérique Droits de l’homme et vie privée à l’âge de la surveillance : le pouvoir du droit international Le droit à la vie privée et le droit à la protection des données La position américaine au regard du droit international relatif aux droits de l’homme et l’initiative germano-brésilienne Renseignement, démocratie, souveraineté : quel demos pour quelle société ? Subjectivité et surveillance du cyberespace Vivre avec la surveillance : résignation, perplexité et résistance

The article identifies how the nexus between democracy, security, humanitarianism and development was built up from the 1990s. It analyses how the discourse of post-conflict peacebuilding has emerged as a notable component of a liberal democratic international order. The article argues that the transformations in peacekeeping operations depend upon a specific spatiotemporal combination – a cleavage between a global and a humanitarian space and the temporality of development. For South American countries, participation in peacekeeping operations became a way to assert themselves as participants of a liberal democratic international order and a reflexive mode to strengthen the process of transformation of their own societies in order to be integrated into a new global cartography.

Based on small scale studies or on little sensitive serological tests, bovines in the south of the state of Rio Grande do Sul, Brazil, are known to be infected with either bovine herpesvirus 1 (BoHV-1) or 5 (BoHV-5). However, whether the prevalence of each of these viruses is high or low is currently still unknown. In order to determine the extent of BoHV (-1 and/or -5) infections in bovines in this region of Brazil, 200 bovines were studied for the presence of BoHV DNA. To this end, first a quantitative PCR was developed that amplified BoHV-1 DNA as well as BoHV-5 DNA. Using this PCR the number of BoHV genomes normally present in latently infected ganglia of naturally infected bovines was estimated. The new PCR was sensitive enough to detect most BoHV DNA in infected ganglia. The results of this first PCR showed that at least 87% of the bovines in the south of Rio Grande do Sul were (latently) infected with either BoHV-1 or BoHV-5. To determine the prevalence of BoHV-1 and BoHV-5 separately, two type-specific PCRs - one for each virus - were developed that used the products of the first PCR as a template. The results of these type-specific PCRs showed that 82.8% of the BoHV positive population was (latently) infected with BoHV-1, 93.1% with BoHV-5 and 75.9% with both BoHV-1 and BoHV-5. This is the first time that such a high frequency of co-infection of BoHV-1 and BoHV-5 in bovines has been demonstrated.

In the present study whole genome of six Brazilian isolates of PCV2 were sequenced, analyzed and compared with 35 other sequences (24 from other countries and 17 from Brazil). The phylogenetic analysis showed that mostly Brazilian variants of PCV2 were grouped as PCV2-1. Two isolates among the six analyzed here could not be grouped with any other PCV2-2 analyzed in this study. One of these isolates was from an aborted fetus with myocarditis and the other from a PMWS affected pig. The results pointed here showed that both groups of PCV2 are present in Brazilian pig population without any clear geographical correlation.

Different types and subtypes of bovine herpesvirus 1 and 5 (BoHV-1 and BoHV-5) have been associated to different clinical conditions of cattle, in such a way that type/subtype differentiation has become an essential tool for understanding the pathogenesis and epidemiology of BoHV infections. In search for a genomic region that would allow a clear distinction between BoHV-1 and BoHV-5, the carboxy-terminal portion of glycoprotein C (gC), corresponding to residues 321–450 (BoHV-1) and 301–429 (BoHV-5) of 23 South American (SA) isolates (Brazil mostly) was amplified and sequenced. The nucleotide sequence alignments revealed levels of genomic similarity ranging from 98.7 to 99.8% among BoHV-1 isolates, 88.3 to 92% between BoHV-1/BoHV-5 and 96 to 99.7% among BoHV-5 isolates. At the amino acid level, sequence similarity varied ranging from 97.5 to 99.5% among BoHV-1, 77.5 to 84.4% between BoHV-1/BoHV-5 and 92.1 to 99.5% (BoHV-5/BoHV-5). The isolates could be clearly separated into BoHV-1.1, BoHV-1.2 and BoHV-5 after phylogenetic analysis. The results suggest that the phylogenetic analysis performed here can be used as a potential molecular epidemiological tool for herpesviruses.

Aujeszky' s disease (AD) is an infectious disease causing important economic losses to the swine industry worldwide. The disease is caused by an alpha-herpesvirus, Aujeszky' s disease virus (ADV), an enveloped virus with a double stranded linear DNA genome. The ADV genome encodes 11 glycoproteins, which are major targets for the immune system of the host in response to the infection. The glycoprotein E (gE) is a non-essential protein and deletion of the gE gene has been used for the production of marker vaccines. Aiming to develop molecular reagents for the production of a gE specific ELISA test, the gE gene was amplified by PCR, cloned and expressed into a baculovirus expression system. The recombinant DNA vector pFastBac-gE-ADV was used for site-specific transposition into the recombinant baculovirus (bacmid). Colonies with recombinant bacmid-pFastBac-gE-ADV were selected by antibiotic and color selection and the presence of the gE gene was confirmed by PCR. The recombinant bacmid-pFastBac-gE-ADV was cotransfected in insect Trichoplusia ni and the presence of the recombinant DNA and gE protein were detected by PCR, SDS-PAGE and Western blotting, respectively.A doença de Aujeszky (DA) é uma enfermidade infecto-contagiosa que causa grandes perdas econômicas ao produtor e à agroindústria suinícola em todo o mundo. É causada pelo vírus da doença de Aujeszky (VDA), um alfaherpesvírus envelopado com genoma DNA de fita dupla e linear. O genoma do VDA codifica 11 glicoproteínas, as quais são os maiores alvos do sistema imune do hospedeiro em resposta a infecção. A glicoproteína E (gE) é uma proteína não essencial e a deleção do gene da gE é muito utilizada para a produção de vacinas com marcadores. Com o objetivo de desenvolver insumos moleculares para a produção de um teste de ELISA específico para gE do VDA, a seqüência do gene da gE foi amplificada, clonada e expressa no sistema de expressão em baculovírus. O produto da amplificação foi clonado no vetor pGem®-T Easy e subclonado no plasmídeo de expressão pFastBac™1. O DNA recombinante pFastBac-gE-VDA foi usado para a transposição sítio-específica no baculovírus recombinante (bacmid). Após seleção por antibióticos e cor, as colônias com o recombinante bacmid-pFastBac-gE-VDA foram selecionadas e a presença do gene da gE foi confirmada por PCR. O DNA recombinante viral, bacmid-pFastBac-gE-VDA, foi usado para cotransfecção de células de inseto Trichoplusia ni e a presença do recombinante e a proteína gE foi determinada por PCR, por SDS-PAGE e Western blotting, respectivamente.

A última década foi palco de importantes transformações no campo da Cooperação Internacional para o Desenvolvimento (CID). Nesse período, novos provedores - países em desenvolvimento, agentes privados e organizações civis - tornaram-se protagonistas no campo. Entre os "novos provedores", os países BRICS ocupam posição de destaque. Este artigo discute as transformações estruturais do campo da CID e, particularmente, os efeitos da emergência de novos provedores, como os BRICS, sobre as posições e as dinâmicas que caracterizam o campo. Pretende demonstrar que a emergência de "novos provedores" e de novas modalidades de cooperação internacional resultou no descentramento do campo da CID. Nesse contexto, abre-se um processo de reordenamento do campo no qual a arquitetura institucional da CID se torna palco de intensa disputa política entre doadores tradicionais e "novos provedores". O artigo analisa alguns aspectos centrais dessa disputa e sugere que o processo, em curso, de descentramento e reordenamento do campo da CID desestabilizou não apenas a posição ocupada pelos doadores tradicionais, como também a posição dos novos provedores. The past decade was the stage of major transformations in the field of International Cooperation for Development (ICD). In this period, new providers - developing countries, private agents and civil organizations - became leading figures in the field. Among those "new providers", the BRICS countries occupy a prominent position. This article discusses the structural transformations of the ICD field and, particularly, the effects of the emergence of new providers, such as the BRICS, on the positions and dynamics that characterize the field. The article intends to demonstrate that the emergence of "new providers" and new modalities of international cooperation resulted in the decentralization of the field of ICD. In this context, begins a process of reordering of the field, in which the institutional architecture of the ICD becomes a stage of intense political quarrel between traditional donors and "new providers". The article analyses some central features of this quarrel and suggests that the ongoing process of decentering and reordering of the field of ICD destabilized not only the position occupied by traditional donors, but also the position of the new providers.

During a series of experiments attempting to reproduce clinically apparent bovine herpesvirus type 5 (BoHV-5) infections, a group of calves was inadvertently infected with bovine viral diarrhoea virus (BVDV). Another group of calves was infected with BoHV-5 only. This paper reports the outcome of such infections. Two out of six calves solely infected with BoHV-5 displayed moderate to severe clinical signs. Three out of four calves of the group co-infected with BoHV-5 and BVDV developed severe clinical signs, two of them died. BoHV-5 virus was isolated to higher titres and for a longer period from the group of calves infected with both viruses. These results suggest that BVDV may enhance clinical signs induced by BoHV-5 and may also play a role in extending the period of BoHV-5 shedding.

In the present study, cross-protection to bovine herpesvirus type 5 (BHV-5) induced by bovine herpesvirus type 1 (BHV-1) vaccination was examined following inoculation of rabbits and calves with a glycoprotein E (gE)-negative BHV-1 vaccine and subsequent challenge with BHV-5. Rabbits (n=5) and calves (n=8) were vaccinated [five rabbits intranasally (IN), four calves IN and four intramuscularly (IM)] with 7.1 log(10)median tissue culture infective dose (TCID(50)) of the BHV-1 vaccine. Rabbits and calves were challenged IN [rabbits 2 weeks post-vaccination (pv); calves 5 weeks pv] with 9.1log(10)TCID(50) of BHV-5. Two out of five vaccinated rabbits died after challenge with typical BHV-5 disease, as did 3/5 non-vaccinated controls. In calves, 4/8 vaccinated animals displayed mild signs of disease, whereas 6/6 non-vaccinated controls developed signs of disease, so severe that 2/6 had to be killed. Besides, nasal virus shedding post-challenge was not reduced by vaccination. At necropsy, on day 21 post-challenge, typical BHV-5 lesions were evident in brain tissues of both vaccinated and non-vaccinated calves. Dexametasone administration at 180 days post-infection did not reactivate clinical signs despite BHV-5 shedding in nasal secretions of both vaccinated and non-vaccinated calves. These results show that the BHV-1 vaccine evaluated here did not confer protection to BHV-5 in rabbits. In calves, BHV-1 vaccination did confer some protection to BHV-5 induced clinical disease, but it did not prevent infection and had no effect on nasal virus shedding or on the development of encephalitic lesions.

Bovine herpesvirus type 5 (BHV-5) is the causative agent of bovine herpetic encephalitis, a major concern for cattle farming in Brazil and Argentina. We recently developed a differential, gE-negative vaccine (265 gE-), based on a Brazilian BHV-1 strain. The present study was carried out to examine whether such a vaccine would confer protection to BHV-5 infections. It was concluded that the recombinant BHV-1 vaccine tested here is not capable of conferring full protection to BHV-5 challenge.

This paper presents data collected by a Brazilian center in a multinational multicenter observational study of patients with mucopolysaccharidosis type VI (MPS VI), aiming at determining the epidemiological, clinical, and biochemical profile of these patients. Twenty-eight south-American patients with MPS VI were evaluated through medical interview, physical exam, echocardiogram, electrocardiogram, ophthalmologic evaluation, quantification of glycosaminoglycans (GAGs) in urine, and measurement of the activity of N-acetylgalactosamine-4-sulfatase (ARSB) in leukocytes. 92.9% of patients were Brazilian. Mean age at diagnosis and at evaluation was 48.4 months and 97.1 months, respectively. 88% of patients had onset of symptomatology before the age of 36 months. Consanguinity was reported by 27% of the families. Mean weight and height at birth were 3.481 kg and 51.3 cm, respectively. The most frequently reported clinical manifestations were short stature, corneal clouding, coarse facial features, joint contractures, and claw hands. All patients presented with echocardiogram changes as well as corneal clouding. Mean ARSB activity in leukocytes was 5.4 nmoles/h/mg protein (reference values: 72-174), and urinary excretion of GAGs was on average 7.9 times higher than normal. The number of clinical manifestations did not show a significant correlation with the levels of urinary GAGs nor with the ARSB activity. Also, no significant correlation was found between the levels of urinary GAGs and the ARSB activity. It was concluded that MPS VI has high morbidity and that, when compared with data published in the literature, patients in our study were diagnosed later and presented with a higher frequency of cardiological findings.

O encéfalo de um bezerro macho, de um ano de idade, que morreu com sinais de doença neurológica, foi submetido ao laboratório para diagnóstico de raiva. O exame histopatológico revelou mielite moderada, meningoencefalite leve com infiltração perivascular e corpúsculos de Negri nas células de Purkinje do cerebelo. A infecção pelo vírus rábico foi ainda confirmada por imunofluorescência direta e por inoculação em camundongos. Além disso, um herpesvírus foi isolado dos tecidos do encéfalo. O vírus isolado foi antigênica e geneticamente caracterizado como Herpesvírus bovino tipo 5 (BHV-5). Não foi possível determinar se o BHV-5 teve algum papel ativo no desfecho da enfermidade, uma vez que o vírus poderia estar presente em forma latente nos tecidos neurais. Este é o primeiro relato de uma infecção mista pelo vírus da raiva e BHV-5 em bovinos. The brain of an one year old male calf which died with signs of neurological disease was submitted to the laboratory for rabies diagnosis. Microscopical findings included moderate mielitis, mild meningoencephalitis with perivascular cell cuffing and Negri inclusion bodies in Purkinje cells of the cerebellum. Rabies virus infection was further confirmed by the direct fluorescent antibody test as well as by mouse inoculation. In addition, a herpesvirus was isolated from brain tissues. The isolate was antigenic and genetically characterized as bovine herpesvirus type 5 (BHV-5). It was not possible to determine whether BHV-5 played an active role in the outcome of the infection, since, the virus might have been present in a latent form in neural tissues. This is the first report of a mixed rabies/ BHV-5 infection in calves.

O perfil antigênico de 45 herpesvírus (44 de bovinos, sendo seis amostras de referência de BHV-1 e 15 prováveis BHV-1; três amostras de referência de BHV-5 e 20 prováveis BHV-5) e uma amostra de herpesvírus bubalino (BuHV) foi examinado com um painel de anticorpos monoclonais (Acms) produzidos contra antígenos de herpesvírus bovinos. Para os exames, foi utilizada a prova de imunoperoxidase (IPX) sobre cultivos de células infectadas, tendo os Acms como anticorpos primários. A determinação dos padrões de reatividade das amostras de vírus frente aos Acms permitiu a diferenciação entre os tipos 1 e 5. Todas as amostras isoladas de casos de encefalite apresentaram perfil de BHV-5. Quatro amostras de BHV-5 isoladas de áreas geograficamente distintas apresentaram perfís de reatividade diferenciados em relação às demais amostras do tipo 5. Duas amostras de vírus com perfil antigênico de BHV-5 foram isoladas de sêmen de animais infectados. Estes resultados comprovam a utilidade da caracterização antigênica com este painel de Acms na tipagem de amostras de BHV-1 e BHV-5.

The use of monoclonal antibodies (MAbs) has contributed to the diagnosis of avian infectious bronchitis (IB), however their use are still limited because the utilized MAbs are imported, increasing the cost of such methodologies. Therefore, given the relevance of the national poultry production as well as the losses caused by the occurrence of IB in commercial poultry flocks, it is important that Brazil can produce biological supplies to use for diagnostic of IB. Thus, the present work reports the production, and characterization of MAbs against the infectious bronchitis virus (IBV). Two groups of three Balb-c female mice were inoculated with inactivated classic or variant IBV for 10 times, once per week. The splenocytes from mouse with higher serum concentration against IBV antigens were harvested and fused with murine Sp2/O-Ag14 myeloma cells. We generated two hybridomas secreting MAbs anti-IBV (MAb 2G4 and 5A11) from the fusion of myeloma cells and B-lymphocytes from a mouse immunized with the variant IBV. Following the characterization by ELISA and Western Blot, it was observed that MAbs recognize the IBV classic and variant types, showing more intense reactions with IBV variant. The MAbs obtained in this study have potential for use in immunodiagnostic for IB.

Current revelations about the secret US-NSA program, PRISM, have confirmed the large-scale mass surveillance of the telecommunication and electronic messages of governments, companies, and citizens, including the United States' closest allies in Europe and Latin America. The transnational ramifications of surveillance call for a re-evaluation of contemporary world politics' practices. The debate cannot be limited to the United States versus the rest of the world or to surveillance versus privacy; much more is at stake. This collective article briefly describes the specificities of cyber mass surveillance, including its mix of the practices of intelligence services and those of private companies providing services around the world. It then investigates the impact of these practices on national security, diplomacy, human rights, democracy, subjectivity, and obedience.

Immunoglobulin Y (IgY) is the major antibody isotype in birds, reptiles, amphibia, and lungfish, playing a similar biological role as mammal IgG. Due to its phylogenetic distance, immune diversification and presence in the egg yolk, IgY provide a number of advantages in immunodiagnostic compared to IgG from mammals. Moreover, IgY production is in agreement with international efforts to reduce, refine and if possible, to replace animals in experimentation, contributing substantially in favor of animal welfare. This article presents an overview about structural and functional features, production and applications of IgY in immunodiagnostic, as well as the advantages of chicken antibodies use. A imunoglobulina Y (IgY) é a classe de anticorpos de maior importância em aves, répteis, anfíbios e peixes pulmonados, desempenhando um papel semelhante a IgG de mamíferos. Devido a sua distância filogenética, mecanismos de diversificação imune e presença na gema do ovo, IgY proporciona uma série de vantagens em imunodiagnóstico, quando comparada a IgG de mamíferos. Além disso, esse método alternativo de produção de anticorpo está de acordo com os esforços internacionais para reduzir, refinar e, se possível, substituir animais em experimentação, contribuindo substancialmente a favor do bem-estar animal. Este artigo apresenta uma revisão sobre as características estruturais e funcionais da IgY, bem como os métodos de produção, vantagens e aplicações em imunodiagnóstico, além das vantagens da sua utilização.

Um ensaio imunoenzimático do tipo ELISA de bloqueio com anticorpo monoclonal (ELISA-M) foi desenvolvido e padronizado para a detecção de anticorpos contra o vírus da Rinotraqueíte Infecciosa Bovina (Herpesvírus Bovino tipo 1; BHV-1). Foram utilizadas nesta avaliação 266 amostras de soros bovinos, sendo 148 negativos e 118 positivos em testes de soroneutralização (SN). Em comparação com este último, o ELISA-M demonstrou uma sensibilidade de 92,37%, especificidade de 92,56%, valor preditivo positivo de 90,83%, valor preditivo negativo de 93,83% e precisão de 92,48%. O índice de concordância (k) entre os testes foi de 0,85. O ELISA-M apresentou como vantagens a rapidez e a praticidade de execução. Com base nestes resultados, o ELISA-M foi considerado uma alternativa apropriada para o diagnóstico sorológico de infecções pelo BHV-1. Entretanto, o teste não foi capaz de diferenciar anticorpos induzidos por BHV-1 ou BHV-5.

The article examines one of the strategies of national identification that characterized nineteenth-century Brazilian fiction and literary criticism. This prevalent line — which the author has designated "exotic trope" — employed a nationalist rhetoric and was a pre-figurative movement containing within itself two separate, complementary movements. The first of these saw the nation as an expressive whole and sought to uncover its historical essence. The second endeavored to identify the nation and its destination vis-a-vis universal civilization. Within nineteenth-century fiction and criticism, the exotic trope movement was impermeable to criticism and gave rise to various forms of national imagination

The international system in general, and the international cooperation for development specifically, have been through important changes in the last decades. The emergence of South-South Cooperation (SSC) has become a trending topic among academics, practitioners and policy-makers. The assumption that the common problems and shared experiences of countries in the global South would make SSC more legitimate and perhaps more effective is frequently mentioned, as a former Brazilian minister of foreign affairs once said, “for every African problem there is a Brazilian solution”. This paper challenges this assertion synthesizing key findings, contextual information and analysis required for understanding Brazil’s engagement in Angola, within the sector of public health, from 2006 to 2015.