Schistosomiasis epidemiology and control: how did we get here and where should we go?

Although a disease of great antiquity, scientific studies of schistosomiasis began only 150 years ago. The complete life-cycle was not described until just before the First World War, making it possible at last to plan proper community control programmes. Inadequate tools prevented their effective implementation until well after the Second World War when new tools became available, thanks to the newly formed World Health Organization. Molluscicides spearheaded control programmes until the late 1970s but were then replaced by the newly developed, safe drugs still used today. Whatever the method used, the initial goal of eradication was, in the light of experience and cost, gradually replaced by less ambitious targets; first to stop transmission and then to reduce morbidity. The most successful programmes combined several methods to minimise reinfection after chemotherapy. Comparisons between different programmes are difficult without using appropriate, standardised diagnostic techniques and the correct epidemiological measurements. Some examples will be presented, mainly from our studies on Schistosoma mansoni in Kenya. Drug resistance on a scale comparable with malaria has not occurred in schistosomiasis but the likely withdrawal of all drugs except praziquantel leaves its control extremely vulnerable to this potential problem. An effective, affordable vaccine for use in endemic countries is unlikely to be ready for at least 5 years, and developing strategies for its use could take a further decade or more, judging from experience with drugs and molluscicides. In the interim, by analogy with malaria, the most cost-effective approach would the use of drugs combined with other methods to stop transmission, including molluscicides. The cost of molluscicides needs to be reduced and fears allayed about their supposedly adverse ecological effects.

Screening of some plants used in the Brazilian folk medicine for the treatment of infectious diseases

Extracts of 13 Brazilian medicinal plants were screened for their antimicrobial activity against bacteria and yeasts. Of these, 10 plant extracts showed varied levels of antibacterial activity. Piper regnellii presented a good activity against Staphylococus aureus and Bacillus subtilis, a moderate activity on Pseudomonas aeruginosa, and a weak activity against Escherichia coli. Punica granatum showed good activity on S. aureus and was inactive against the other standard strains. Eugenia uniflora presented moderate activity on both S. aureus and E. coli. Psidium guajava,Tanacetum vulgare, Arctium lappa, Mikania glomerata, Sambucus canadensis, Plantago major and Erythrina speciosa presented some degree of antibacterial activity. Spilanthes acmella, Lippia alba, and Achillea millefolium were considered inactive. Five of the plant extracts presented compounds with Rf values similar to the antibacterial compounds visible on bioautogram. Of these, three plants belong to the Asteraceae family. This may mean that the same compounds are responsible for the antibacterial activity in these plants. Anticandidal activity was detected in nine plant extracts (P. guajava, E. uniflora, P. granatum, A. lappa, T. vulgare, M. glomerata, L. alba, P. regnellii, and P. major). The results might explain the ethnobotanical use of the studied species for the treatment of various infectious diseases.

Oviposition response of the mosquito, Culex quinquefasciatus to the secondary metabolite(s) of the fungus, Trichoderma viride

Secondary metabolites produced by Trichoderma viride, a deuteromycetes fungus, under submerged culture condition were formulated and evaluated for oviposition attractancy against gravid females of Culex quinquefasciatus mosquito. At a concentration of 10 µg ml-1 the formulation showed remarkable attractancy with an oviposition active index (OAI) of +0.52. When the oviposition attractancy of the formulation was compared with a known oviposition attractant, p-cresol, both at 10 µg ml-1, the former was found to be more attractive to result in 70% egg laying than the later with 30% egg laying. Thin layer chromatography fractions of the secondary metabolites showed that a fraction with Rf value of 0.88 was highly active as oviposition attractant with an OAI of +0.65. Further work on identification of the active principle(s) of the microbial formulation might lead to an oviposition attractant useful in mosquito vector management.

Prevalence of human papillomavirus types in women with pre-neoplastic and neoplastic cervical lesions in the Federal District of Brazil

As a contribution to the public health authorities in planning prophylactic and therapeutic vaccine strategies, we describe the prevalence of human papillomavirus (HPV) types in women presenting abnormal cytological results in Pap smear screening tests in the Federal District, Central Brazil. We studied 129 cervical scraping samples from women whose cytological tests showed either pre-neoplastic or neoplastic lesions. Amplification of HPV DNA was performed by polymerase chain reaction using consensus primers MY09 and MY11 followed by identification of isolates by restriction fragment length polymorphism. We detected HPV DNA in 62% of the samples, including HPV-16 in 43.8%, HPV-58 in 12.5%, HPV-31 in 10%, HPV-53 in 6.3%, each of HPV-18 and HPV-33 in 3.8% of the isolates. Other types (HPV-35, -52, -66, -CP8304, -6, -11, and -CP8061) were less frequent (= or < 2.5% each). The prevalence of HPV-58 was relatively higher in this population than in data in South America, but similar to results obtained in other studies in Latin America, Europe, and Eastern Asia. Case-control studies need to be carried out to establish the association between the prevalence of HPV types specially the less frequent high-risk types and cervical cancer.

Prevalence of human papillomavirus type 16 variants in the Federal District, Central Brazil

We report the prevalence of human papillomavirus type 16 (HPV-16) variants in women with cervical lesions from the Federal District, Central Brazil. We analyzed 34 HPV-16 samples, identifying the sequence variations of E6 and L1 genes and correlating variant frequency with disease status. The most prevalent HPV-16 variant was the European (50%), followed by Asian-American (41.2%), African-1 (5.9%), and African-2 (2.9%). European and non-European variants appeared in equal frequencies among the cytological types of lesions - atypical squamous or glandular cells of undetermined significance, cytological alterations suggesting HPV infection, cervical intraepithelial neoplasias, squamous cell carcinoma, and adenocarcinoma.

Antiretroviral resistance and genetic diversity of human immunodeficiency virus type 1 isolates from the Federal District, Central Brazil

In the context of universal access to antiretroviral therapy, the surveillance of human immunodeficiency virus type 1 (HIV-1) genetic diversity and resistance becomes pivotal. In this work our purpose was to describe the genetic variability; prevalence of drug-resistance mutations; and genotypic resistance profiles in HIV-1 infected individuals under antiretroviral treatment, from the Federal District, Brasília, Central Brazil. The entire viral protease and codons 19 to 234 of the reverse transcriptase gene from 45 HIV-1 isolates were amplified and sequenced for subtyping and genotyping. By phylogenetic analysis, 96% of the samples clustered with subtype B and the remaining 4% with HIV-1 subtype F sequences. One major protease inhibitor resistance-associated mutation, I50V, was detected in 38% of the samples. Minor mutations were also found at the protease gene: L10I/V (7%), K20M (2%), M36I (11%), L63P (20%), A71T (2%), and V77I (7%). Many mutations associated with reduced susceptibility to nucleoside or non-nucleoside reverse transcriptase inhibitors were detected: M41L (11%), E44D (4%), D67N (11%), T69D (2%), K70R (11%), L74V (2%), L100I (4%), K103N (18%), V118I (9%), Y181C (11%), M184V (18%), G190A (4%), T215Y (4%), and K219E (4%). This study has shown that 84% of the studied population from the Federal District, showing evidences of therapy failure, presented viral genomic mutations associated with drug resistance. The main antiretrovirals to which this population showed resistance were the PI amprenavir (38%), the NNRTIs delavirdine, nevirapine (31%), and efavirenz (24%), and the NRTIs lamivudine (18%), abacavir, and zidovudine (13%).

Hepatitis C virus genotypes in blood donors from the Federal District, Central Brazil

The objective of this study was to characterize hepatitis C virus (HCV) genotypes in blood donors from the Federal District, Central Brazil, and to compare HCV screening by serological assays and reverse transcriptase polymerase chain reaction (RT-PCR). Plasma samples from 57 individuals with reactive or indeterminate results in serological anti-HCV screening assays (ELISA or EIA) were tested for HCV RNA by RT-PCR. The results from a confirmatory LIA serological assay were also evaluated. The 5' non-coding region of the HCV genome was amplified from 41 PCR positive samples (71.9%), which were further characterized by nucleotide sequencing analysis. Of these, 60.9% were of HCV genotype 1 and 39.1% of genotype 3.

In vitro inhibition of Plasmodium falciparum by substances isolated from Amazonian antimalarial plants

In the present study, a quassinoid, neosergeolide, isolated from the roots and stems of Picrolemma sprucei (Simaroubaceae), the indole alkaloids ellipticine and aspidocarpine, isolated from the bark of Aspidosperma vargasii and A. desmanthum (Apocynaceae), respectively, and 4-nerolidylcatechol, isolated from the roots of Pothomorphe peltata (Piperaceae), all presented significant in vitro inhibition (more active than quinine and chloroquine) of the multi-drug resistant K1 strain of Plasmodium falciparum. Neosergeolide presented activity in the nanomolar range. This is the first report on the antimalarial activity of these known, natural compounds. This is also the first report on the isolation of aspidocarpine from A. desmanthum. These compounds are good candidates for pre-clinical tests as novel lead structures with the aim of finding new antimalarial prototypes and lend support to the traditional use of the plants from which these compounds are derived.

Evolução quaternária, distribuição de partículas nos solos e ambientes de sedimentação em manguezais do estado de São Paulo

A distribuição de partículas em solos ou sedimentos das planícies litorâneas auxilia no entendimento dos processos de sedimentação em estuários, servindo com importante atributo para aplicações em estudos de reconstrução paleoambiental, ciclos geoquímicos e poluição ambiental, como contaminação por metais pesados e derrames de petróleo, que, devido à ação antrópica, são relativamente comuns nesses ambientes. Com o objetivo de caracterizar os ambientes de sedimentação de acordo com a granulometria e com o processo de evolução quaternária ao longo do litoral do Estado de São Paulo, foram estudados solos de 14 manguezais. As análises granulométricas foram realizadas nas camadas de 0-20 e 60-80 cm de profundidade, determinando as frações argila, silte, areia total e cinco frações da areia. Realizaram-se datações 14C por cintilação líquida, espectrometria de massa acoplada a acelerador de partículas na fração humina da matéria orgânica e por termoluminescência em grãos de quartzo, para amostras de diferentes camadas dos manguezais amostrados. Os resultados de granulometria foram tratados de acordo com os parâmetros estatísticos de Folk & Ward. Os solos dos manguezais do Estado de São Paulo têm idade holocênica oscilando entre 410 e 3.700 anos AP, até a profundidade de 80 cm. Em alguns casos este substrato holocênico encontra-se sobreposto à camada arenosa pleistocênica, como foi identificado em SG1 (65-77 cm = 11.000 anos e 90-95 cm = 24.700 anos), PM (72-79 cm = 60.000 anos) e em RF, cuja camada a 40-50 cm apresentou idade de 12.200 anos. Os manguezais apresentam solos de diferentes texturas, variando de arenosa a argilosa. Os solos de constituição arenosa foram identificados na Ilha do Cardoso, na planície do Rio Guaratuba e ao longo do litoral norte, cujos manguezais foram estabelecidos sobre os sedimentos retrabalhados de antigos cordões arenosos e localizados às margens dos rios que drenam essas planícies litorâneas (SG1, SG2, GUA e RE). Esses solos ocorrem também nas proximidades da desembocadura dos rios, onde há maior influência da ação de ondas (RF). Os manguezais cujo substrato são constituídos, predominantemente, de partículas finas (silte e argila) estão localizados nos compartimentos mais protegidos do litoral, como o Canal do Cananéia (PM e BAG), Mar Pequeno (IGUA) e dentro do estuário de Santos, na Baixada Santista (ITA, IRI, COS e CRU). O período de maré estacionada, que decorre da alternância dos ciclos de enchente e vazante desta, favoreceria a sedimentação de partículas da fração silte, explicando a ocorrência de alto teor de silte na superfície dos solos de manguezais de PM e BAG e ao longo das camadas estudadas de IGUA e ITA.

Análise multivariada de atributos microbiológicos e químicos do solo em florestas com Araucaria angustifolia

Araucaria angustifolia é uma espécie de árvore ameaçada de extinção no Brasil e pouco se conhece sobre os atributos edáficos dessas florestas. Este estudo foi realizado com o objetivo de identificar diferenças entre áreas com araucária naturais e reflorestadas, com base em atributos microbiológicos e químicos do solo, por meio de métodos multivariados, como a análise canônica discriminante (ACD) e a análise de correlação canônica (ACC). As áreas estudadas incluem: 1. floresta nativa com araucária (NF); 2. reflorestamento de araucária (R); 3. reflorestamento de araucária submetido a incêndio acidental (RF); e 4. pastagem natural com araucárias nativas e ocorrência de incêndio (NPF). Foram selecionadas, ao acaso, quinze árvores de araucária por área e sob a copa de cada uma delas foram retiradas três amostras de solo, em três épocas contrastantes. A ACD foi aplicada aos atributos microbianos: C da biomassa microbiana (CBM), respiração basal (C-CO2) e quociente metabólico (qCO2), enquanto a ACC foi aplicada aos atributos microbianos e químicos do solo [pH (CaCl2), C orgânico total (COT) e teores de P, K, Ca, Mg e (H+Al). Os atributos microbianos e químicos do solo apresentaram alta correlação canônica, com destaque para o CBM e C-CO2, entre os atributos microbianos, e para COT e P, entre os atributos químicos do solo. A ACC mostrou que 52 % da variação total dos atributos microbianos é explicada pela variação dos atributos químicos, e 36 % da variação total dos atributos químicos é explicada pelos atributos microbianos do solo das áreas estudadas. A ACD identificou que a contribuição de cada atributo microbiano para a separação das áreas sofreu variação ao longo do tempo, e discriminou o CBM como sendo o atributo microbiano mais importante, seguido pelo C-CO2. ACC e ACD são importantes ferramentas no estudo de indicadores de qualidade do solo.

Colêmbolos (Hexapoda: Collembola) como bioindicadores de qualidade do solo em áreas com Araucaria angustifolia

Não existem informações sobre a diversidade de colêmbolos associados às florestas com Araucaria angustifolia (Bert.) O. Kuntze no Brasil. Este estudo teve o objetivo de avaliar o potencial da diversidade de famílias de colêmbolos como bioindicadores da qualidade do solo e para separar diferentes áreas com araucária, utilizando atributos químicos e microbiológicos do solo como variáveis explicativas. O estudo foi realizado em quatro áreas: 1. floresta nativa com predominância de araucária (NF); 2. reflorestamento de araucária (R); 3. reflorestamento de araucária submetido a incêndio acidental (RF); e 4. pastagem natural com araucárias nativas e ocorrência de incêndio acidental (NPF). Na captura dos organismos, utilizaram-se dez armadilhas, distribuídas ao acaso em cada área, nos mesmos pontos de coleta dos atributos químicos e microbiológicos do solo. O número de colêmbolos de cada família, extraído por armadilhas (abundância), o índice de diversidade de Shannon (H) e a riqueza de famílias foram submetidos à Análise de Variância (ANOVA). A abundância de cada família de Collembola foi submetida à Análise de Componentes Principais (ACP). Considerando as quatro áreas, foram identificadas oito famílias de colêmbolos (Brachystomellidae, Entomobryidae, Hypogastruridae, Isotomidae, Katiannidae, Paronellidae, Sminthuridae e Tomoceridae). A diversidade de famílias de colêmbolos foi maior nas áreas NF e R, em comparação com as áreas RF e NPF. Os atributos químicos e microbiológicos do solo foram eficientes como variáveis explicativas, pois auxiliaram na interpretação das modificações das famílias de colêmbolos. A ACP mostrou que a identificação ao nível de família de Collembola é suficiente para separar as áreas estudadas, além de indicar quais foram as famílias de colêmbolos mais associadas a cada área, o que possibilita sua utilização como bioindicadores de intervenções antrópicas, bem como da qualidade do solo.

Water retention in a peatland with organic matter in different decomposition stages

Peatlands are ecosystems formed by successive pedogenetic processes, resulting in progressive accumulation of plant remains in the soil column under conditions that inhibit the activity of most microbial decomposers. In Diamantina, state of Minas Gerais, Brazil, a peatland is located at 1366 m asl, in a region with a quartz-rich lithology and characteristic wet grassland vegetation. For this study, the peat area was divided in 12 transects, from which a total of 90 soil samples were collected at a distance of 20 m from each other. The properties rubbed fiber content (RF), bulk density (Bd), mineral material (MM), organic matter (OM), moisture (Moi) and maximum water holding capacity (MWHC) were analyzed in all samples. From three selected profiles of this whole area, samples were collected every 27 cm from the soil surface down to a depth of 216 cm. In these samples, moisture was additionally determined at a pressure of 10 kPa (Moi10) or 1500 kPa (Moi1500), using Richards' extractor and soil organic matter was fractionated by standard procedures. The OM decomposition stage of this peat was found to increase with soil depth. Moi and MWHC were highest in layers with less advanced stages of OM decomposition. The humin levels were highest in layers in earlier stages of OM decomposition and with higher levels of water retention at MWHC and Moi10. Humic acid contents were higher in layers at an intermediate stage of decomposition of organic matter and with lowest levels of water retention at MWHC, Moi10 and Moi1500.

Relationships between microbial activity and soil physical and chemical properties in native and reforested Araucaria angustifolia forests in the state of São Paulo, Brazil

Araucaria angustifolia (Bert.) O. Kuntze is the main component of the Mixed Ombrophilous forest and, in the State of São Paulo, it is associated with a high diversity of soil organisms, essential for the maintenance of soil quality, making the conservation of this ecosystem a major and pressing challenge. The objective of this study was to identify the physical and chemical properties that are most closely correlated with dehydrogenase enzyme activity, basal respiration and microbial biomass under native (NF) and replanted (RF) Araucaria angustifolia forests in three regions of the state of São Paulo, in winter and summer. The main differentiating factors between the areas were also determined. Each forest was represented by three true replications; at each site, from around the araucaria trees, 15 soil samples (0-20 cm) were collected to evaluate the soil physical, chemical and microbiological properties. At the same points, forest litter was sampled to assess mass and chemical properties. The following microbiological properties were evaluated: microbial biomass carbon (MBC), basal respiration (CO2-C), metabolic quotient (Q: CO2), dehydrogenase enzyme activity (DHA) as well as the physical properties (moisture, bulk density, macroporosity and total porosity), soil chemical properties [pH, organic carbon (org-C), P, Ca, K, Mg, Al, H+Al], litter dry mass, and C, N and S contents. The data were subjected to analysis of variance (TWO-WAY: ANOVA). A Canonical Discriminant Analysis (CDA) and a Canonical Correlation Analysis (CCA) were also performed. In the soil under NF, the values of K, P, soil macroporosity, and litter dry mass were higher and Q: CO2 and DHA lower, regardless of the sampling period, and DHA was lower in winter. In the RF areas, the levels of moisture, porosity and Q: CO2 were higher in both sampling periods, and DHA was higher in winter. The MBC was only higher under NF in the summer, while the litter contents of C, N and S were greater in winter. In winter, CCA showed a high correlation of DHA with CO2-C, pH and H+Al, while in the summer org-C, moisture, Mg, pH and litter C were more associated with DHA and CO2-C. The CDA indicated H+Al, available P, total porosity, litter S content, and soil moisture as the most discriminating variables between NF and RF, but moisture was the most relevant, in both seasons and CO2-C only in winter. The combined analysis of CCA and CDA showed that the contribution of the microbiological variables to a differentiation of the areas was small at both samplings, which may indicate that the period after reforestation was long enough to allow an almost complete recovery of the microbial activity.

Rice husk ash as corrective of soil acidity

Rice husk ash (RHA) is a by-product from the burning of rice husk that can have favorable effects on the soil in terms of acidity correction. The objectives of this study were to determine the effective calcium carbonate equivalent (ECC) of RHA under field conditions, and establish technical criteria as a basis for estimating the overall ECC of RHA. The 12 treatments of the experiment consisted of 10 RHA dosages (0, 10, 20, 30, 40, 60, 80, 100, 120, and 140 Mg ha-1) and two references, one of which was an absolute control (AC) and the other a plot limed and fertilized according to official recommendations (recommended fertilization - RF). The soil was sampled twice (15 and 210 days after incorporating RHA), in the layers 0.00-0.10 and 0.10-0.20 m, to determine the pH(H2O) and base saturation (V%). The ECC and neutralizing value (NV) of RHA were also determined. The results showed that RHA neutralizes soil acidity, in a faster reaction than conventional limestone, despite a low ECC (around 3 %).