Luiza Terezinha Madia de Souza

Genetic investigation of novel hantaviruses causing fatal HPS in Brazil

Although hantavirus pulmonary syndrome (HPS) was discovered in North America in 1993, more recent investigations have shown that the disease is a much larger problem in South America, where a greater number of cases and HPS‐associated viruses have now been detected. Here we describe the genetic investigation of three fatal HPS cases from Brazil, including a 1995 case in Castelo dos Sonhos (CAS) in the state of Mato Grosso and two 1996 cases in the counties of Araraquara (ARA) and Franca (FRA), in the state of São Paulo. Reverse transcription‐polymerase chain reaction (RT‐PCR) products representing fragments of the hantavirus N, G1, and G2 coding regions were amplified from patient acute‐phase serum samples, and the nucleotide (nt) sequences (394, 259, and 139 nt, respectively) revealed high deduced amino acid sequence identity between ARA and FRA viruses (99.2%, 96.5%, and 100%, respectively). However, amino acid differences of up to 14.0% were observed when ARA and FRA virus sequences were compared with those of the geographically more distant CAS virus. Analysis of a 643‐nt N coding region and a 1734‐nt predominantly G2‐encoding region of ARA and CAS virus genomes confirmed that these Brazilian viruses were distinct and monophyletic with previously characterized Argentinean hantaviruses, and suggested that Laguna Negra (LN) virus from Paraguay was ancestral to both the Brazilian and Argentinean viruses. The phylogenetic tree based on the N coding fragment also placed LN in a separate clade with Rio Mamore virus from Bolivia. At the amino acid level, ARA and CAS viruses appeared more closely related to the Argentinean viruses than they were to each other. Similarly, analysis of the diagnostic 139‐nt G2 fragment showed that the Juquitiba virus detected in a 1993 fatal HPS case close to São Paulo city, Brazil was closer to Argentinean viruses than to ARA or CAS viruses. These data indicate that at least three different hantavirus genetic lineages are associated with Brazilian HPS cases. J. Med. Virol. 59:527–535, 1999.

Identifying Rodent Hantavirus Reservoirs, Brazil

Bolomys lasiurus and Oligoryzomys nigripes are rodent reservoirs of Araraquara-like and Juquitiba-like hantaviruses, which cause HPS in Brazil.

Hantavirus Pulmonary Syndrome, Central Plateau, Southeastern, and Southern Brazil

This syndrome is an increasing health problem because of human encroachment into habitats of rodent reservoirs.

Dengue Virus Type 4 Phylogenetics in Brazil 2011: Looking beyond the Veil

Dengue Fever and Dengue Hemorrhagic Fever are diseases affecting approximately 100 million people/year and are a major concern in developing countries. In the present study, the phylogenetic relationship of six strains of the first autochthonous cases of DENV-4 infection occurred in Sao Paulo State, Parana State and Rio Grande do Sul State, Brazil, 2011 were studied. Nucleotide sequences of the envelope gene were determined and compared with sequences representative of the genotypes I, II, III and Sylvatic for DEN4 retrieved from GenBank. We employed a Bayesian phylogenetic approach to reconstruct the phylogenetic relationships of Brazilian DENV-4 and we estimated evolutionary rates and dates of divergence for DENV-4 found in Brazil in 2011. All samples sequenced in this study were located in Genotype II. The studied strains are monophyletic and our data suggest that they have been evolving separately for at least 4 to 6 years. Our data suggest that the virus might have been present in the region for some time, without being noticed by Health Surveillance Services due to a low level of circulation and a higher prevalence of DENV-1 and DENV- 2.

Surveillance of arbovirus infections in the atlantic forest region, State of São Paulo, Brazil: I. detection of hemagglutination-inhibition antibodies in wild birds between 1978 and 1990

We report data related to arbovirus antibodies detected in wild birds periodically captured from January 1978 to December 1990 in the counties of Salesópolis (Casa Grande Station), Itapetininga and Ribeira Valley, considering the different capture environments. Plasmas were examined using hemagglutination-inhibition (HI) tests. Only monotypic reactions were considered, except for two heterotypic reactions in which a significant difference in titer was observed for a determined virus of the same antigenic group. Among a total of 39,911 birds, 269 birds (0.7%) belonging to 66 species and 22 families were found to have a monotypic reaction for Eastern equine encephalitis (EEE), Venezuelan equine encephalitis (VEE), Western equine encephalitis (WEE), Ilheus (ILH), Rocio (ROC), St. Louis encephalitis (SLE), SP An 71686, or Caraparu (CAR) viruses. Analysis of the data provided information of epidemiologic interest with respect to these agents. Birds with positive serology were distributed among different habitats, with a predominance of unforested habitats. The greatest diversity of positive reactions was observed among species which concentrate in culture fields.

Hantavirus pulmonary syndrome in the State of São Paulo, Brazil, 1993-1998.

Between 1993 and 1998, 10 cases of clinical hantavirus infection were diagnosed in Brazil. Hantavirus-specific IgM, or positive immunohistochemical analysis for hantavirus antigen, or positive reverse transcription-polymerase chain reaction results for hantavirus RNA were used to confirm nine of these cases; eight were hantavirus pulmonary syndrome (HPS), and one was mild hantavirus disease. The remaining clinical case of hantavirus infection was fatal, and no tissue was available to confirm the diagnosis. During the first 7 months of 1998, five fatal HPS cases caused by a Sin Nombre-like virus were reported from three different regions in the State of São Paulo, Brazil: two in March (Presidente Prudente Region), two in May (Ribeirão Preto Region), and one in July (Itapecerica da Serra Region). Epidemiologic, ecologic, and serologic surveys were conducted among case contacts, area residents, and captured rodents in five locations within the State of São Paulo in June of 1998. Six (4.8%) of 125 case contacts and six (5.2%) of 116 area residents had IgG antibody to Sin Nombre virus (SNV) antigen. No case contacts had a history of HPS-compatible illness, and only one area resident reported a previous acute respiratory illness. A total of 403 rodents were captured during 9 nights of trapping (1969 trap nights). All 27 rodents that were found to be positive for IgG antibody to SNV antigen were captured in crop border and extensively deforested agricultural areas where four of the 1998 HPS case-patients had recently worked. The IgG antibody prevalence data for rodents suggest that Bolomys lasiurus and perhaps Akodon sp. are potential hantavirus reservoirs in this state of Brazil.

JUNGLE YELLOW FEVER : CLINICAL AND LABORATORIAL STUDIES EMPHASIZING VIREMIA ON A HUMAN CASE

The authors report the clinical, laboratorial and epidemiological aspects of a human case of jungle yellow fever. The patient suffered from fever, chills, sweating, headaches, backaches, myalgia, epigastric pains, nausea, vomiting, diarrhea and prostration. He was unvaccinated and had been working in areas where cases of jungle yellow fever had been confirmed. Investigations concerning the yellow fever virus were performed. Blood samples were collected on several days in the course of the illness. Three of these samples (those obtained on days 5, 7 and 10) were inoculated into suckling mice in attempt to isolate virus and to titrate the viremia level. Serological surveys were carried out by using the IgM Antibodies Capture Enzyme Linked Immunosorbent Assay (MAC-ELISA), Complement Fixation (CF), Hemagglutination Inhibition (HI) and Neutralization (N) tests. The yellow fever virus, recovered from the two first samples and the virus titration, showed high level of viremia. After that, specific antibodies appeared in all samples. The interval between the end of the viremia and the appearance of the antibodies was associated with the worsening of clinical symptoms, including bleeding of the mucous membrane. One must be aware of the risk of having a urban epidemics in areas where Aedes aegypti is found in high infestation indexes.

Síndrome pulmonar e cardiovascular por hantavírus

Hantavirus pulmonary and cardiovascular syndrome is a recently identified and often fatal disease, which presents as acute respiratory distress syndrome (ARDS). Since the first outbreak, in Nov/Dec 1993, in Juquitiba, Brazil, 226 cases have been registered by FUNASA (National Health Foundation).(4) The disease occurs in previously healthy subjects, presenting with fever and symptoms similar to the common cold, and may rapidly evolve to pulmonary edema, respiratory failure and shock. Hemoconcentration and thrombocytopenia are common features, and the typical radiological finding is a bilateral diffuse interstitial infiltrate that evolves to alveolar consolidations in parallel to the worsening of the clinical condition. Initially, mortality was around 75%, but it declined to approximately 35% in the last few years. Patients who survive usually recover completely, about a week after the onset of the respiratory symptoms. The causal agent is a previously unrecognized hantavirus whose natural reservoirs are rodents of the family Muridae, sub-family Sigmodontinae. Specific antiviral treatment for hantavirus pulmonary and cardiovascular syndrome has not yet been well established, and the efficacy of ribavirin is currently being studied. Intensive care, including mechanical ventilation and invasive hemodynamic monitoring, is required for the more severe presentations of the disease. These measures may improve the prognosis and survival of patients with hantavirus pulmonary and cardiovascular syndrome if started early in the course of the disease.

Soroprevalência da hepatite B e avaliação da resposta imunológica à vacinação contra a hepatite B por via intramuscular e intradérmica em profissionais de um laboratório de saúde pública

OBJECTIVES: To determine the prevalence of HBsAg and anti-HBs and to evaluate the response of intradermal hepatitis vaccination in healthcare workers non-responsive to previous repeated intramuscular vaccination. MATERIAL AND METHOD: All of the employees from Instituto Adolfo Lutz were invited to participate on this study. Serum samples were obtained and HBsAg and anti-HBs were detected using commercial kits (Abbott® Laboratories). Employees were submitted to the conventional three-dose vaccination by intramuscular route. To those employees who did not respond to intramuscular vaccination, 5 µg doses of Engerix® B were then administered by intradermal route up to nine doses. RESULTS: Four hundred and four healthcare workers were enrolled in this study. Initially, two (0.5%) and 42 (10.4%) were HBsAg and anti-HBs reagent, respectively. Among the 360 negative volunteers, 316 (87.8%) received three vaccine doses and in 259 of them, serum samples were collected to evaluate vaccine efficacy. Among them, 242 (93.4%) showed antibodies titer higher than 10 UI/l. Intradermal vaccination was carried out in five volunteers and all of them responded to this vaccine administration route. CONCLUSION: The prevalence of hepatitis B was not higher than in general population. Intradermal vaccine administration could be a good alternative in people that did not respond to previous intramuscular route.

Esquema reduzido de vacinação anti-rábica humana pré-exposição e avaliação de doses anuais de reforço

Sao apresentados os resultados do emprego de esquema de vacinacao anti-rabida humana pre-exposicao, constituido de 3 doses de vacina tipo Fuenzalida-Palacios administradas a 165 pacientes em dias alternados, mais uma dose de reforco no 30.° dia apos a dose inicial. Os titulos de anticorpos foram determinados por prova de soroneutralizacao em amostras de sangue colhidas antes, 30 e 40 dias apos administracao da primeira dose. Verificou-se que no 30.° dia, 74,6% dos pacientes apresentaram anticorpos neutralizantes no soro, valor que se elevou a 98,1% no quadragesimo dia, o que mostra a eficacia do esquema em relacao a resposta imunitaria em tempo relativamente curto e a importância da dose de reforco como estimulo a producao de anticorpos. Nos pacientes submetidos as doses anuais de reforco num periodo de 10 anos, verificou-se aumento gradual da presenca de anticorpos antes da administracao da dose de reforco subsequente, ate atingir valores de 100%. Face aos resultados obtidos foi sugerido que as doses de reforco sejam administradas a intervalos de tempo maiores e precedidas da titulagem de anticorpos a fim de se avaliar da necessidade ou nao de sua administracao.A reduced schedule for a pre-exposure anti-rabic immunization, of humans with 3 doses of 1 ml of Fuenzalida-Palacios type vaccine administered on alternate days, plus one booster 30 days after the first dose, was related. The blood samples were collected on the 0, 30th and 40th day after the first vaccine dose and the antibody titration was performed by serum neutralizing test. It was observed that 74,6% of the patients presented serum neutralizing antibodies on the 30th day and 98.1% on the 40th day, demonstrating the efficacy of this reduced schedule in stimulating the antibody response in a short time. The importance of a booster dose was emphasized. In the patients submitted to annual boosters over a period of 10 years, a gradual increase in the number of patients with anti-rabic antibodies before the subsequent booster dose was observed, reaching the level of 100% in the 9th year. It was suggested that the booster doses be administered at longer intervals and after antibody titration in order to evaluate the need for the administration of the booster dose.