Molecular epidemiology and evolution of HIV-1 in Portugal and portuguese speaking african countries

The aims of this thesis were to better characterize HIV-1 diversity in Portugal, Angola, Mozambique and Cape Verde and to investigate the origin and epidemiological history of HIV-1 in these countries. The impact of these issues in diagnosis, disease progression and susceptibility to ARV therapy was also investigated. Finally, the nature, dynamics and prevalence of transmitted drug resistance (TDR) was determined in untreated HIV-1 infected patients. In Angola, practically all HIV-1 genetic forms were found, including almost all subtypes, untypable (U) strains, CRFs and URFs. Recombinants (first and second generation) were present in 47.1% of the patients. HIV/AIDS epidemic in Angola probably started in 1961, the major cause being the independence war, subsequently spreading to Portugal. In Maputo, 81% of the patients were infected with subtype C viruses. Subtype G, U and recombinants such as CRF37_cpx, were also present. The results suggest that HIV-1 epidemic in Mozambique is evolving rapidly in genetic complexity. In Cape Verde, where HIV-1 and HIV-2 co-circulate, subtype G is the prevailed subtype. Subtypes B, C, F1, U, CRF02_AG and other recombinant strains were also found. HIV-2 isolates belonged to group A, some being closely related to the original ROD isolate. In all three countries numerous new polymorphisms were identified in the RT and PR of HIV-1 viruses. Mutations conferring resistance to the NRTIs or NNRTIs were found in isolates from 2 (2%) patients from Angola, 4 (6%) from Mozambique and 3 (12%) from Cape Verde. None of the isolates containing TDR mutations would be fully sensitive to the standard first-line therapeutic regimens used in these countries. Close surveillance in treated and untreated populations will be crucial to prevent further transmission of drug resistant strains and maximize the efficacy of ARV therapy. In Portugal, investigation of a seronegative case infection with rapid progression to AIDS and death revealed that the patient was infected with a CRF14_BG-like R5-tropic strain selectively transmitted by his seropositive sexual partner. The results suggest a massive infection with a highly aggressive CRF14_BG like strain and/or the presence of an unidentified immunological problem that prevented the formation of HIV-1-specific antibodies. Near full-length genomic sequences obtained from three unrelated patients enabled the first molecular and phylogenomic characterization of CRF14_BG from Portugal; all sequences were strongly related with CRF14_BG Spanish isolates. The mean date of origin of CRF14_BG was estimated to be 1992. We propose that CRF14_BG emerged in Portugal in the early 1990s, spread to Spain in late 1990s as a consequence of IDUs migration and then to the rest of Europe. Most CRF14_BG strains were predicted to use CXCR4 and were associated with rapid CD4 depletion and disease progression. Finally, we provide evidence suggesting that the X4 tropism of CRF14_BG may have resulted from convergent evolution of the V3 loop possibly driven by an effective escape from neutralizing antibody response.

Efeito da Densidade de Estocagen na Sobrevivência e Crescimento do Pampo

Com o intuito de conhecer o potencial zootécnico do pampo, foi avaliado o efeito de diferentes densidades de estocagem (100 e 200 peixes/m3) na sobrevivência e crescimento do pampo amarelo (Trachinotus carolinus), criado em tanque-rede instalado em ambiente marinho. Peixes com aproximadamente 1,0 a 2,0 g, capturados em arrastos nas praias rasas do município de Ubatuba, S.P., foram transportados ao Núcleo de Pesquisa e Desenvolvimento do Litoral Norte – Agência Paulista Tecnológica do Agronegocio – Secretaria da Agricultura e Abastecimento e mantidos durante 45 dias em tanques-rede de 5mm de malhagem. Após seleção manual, animais de 4,0 ± 0,17g foram estocados, nas densidades de 100 e 200 peixes/m3, em tanques-rede de multifilamento com 2 x 2 x 1,5m e 12 mm de malhagem, fixos a um “deck” flutuante (6,0 x 8,0 m), instalado diretamente no mar a profundidade de aproximadamente 3,5 ± 0,5 m. Os animais foram alimentados duas vezes ao dia até a saciação com ração de 40% de PB. Após 120 dias, as taxas de sobrevivência média observadas foram 77,23% e 73,11%; pesos médios (52,79 ± 4.86 g e 52,64 ± 5.64 g); Ganho de Peso (GP) (48,69 ± 4,05 g e 48,73 ± 4,00 g); Taxa de Crescimento Especifico (TCE) (2,13 ± 1,24 e 2,16 ± 1,03) e Conversão Alimentar Aparente (CAA) (3,23 ± 1,52 e 2,66 ± 0,78), respectivamente para 100 e 200 peixes/m3. Estes dados não apresentaram diferenças significativas (P 0,05) entre os tratamentos testados. Entretanto os ganhos de peso do pampo, no presente experimento foram superiores a muitos trabalhos encontrados na literatura, o que nos leva a concluir que a densidade de 200 peixes/m3, utilizada neste estudo, pode ainda estar abaixo da capacidade de suporte para a espécie e pode ser o ponto de partida para futuros estudos, considerando o estágio de desenvolvimento dos animais.