Variation in cuticular hydrocarbons among strains of the Anopheles gambiae sensu stricto by analysis of cuticular hydrocarbons using gas liquid chromatography of larvae

Cuticular hydrocarbons of larvae of individual strains of the Anopheles gambiae sensu stricto were investigated using gas liquid chromatography. Biomedical discriminant analysis involving multivariate statistics suggests that there was clear hydrocarbon difference between the Gambian(G3), the Nigerian (16CSS and, its malathion resistant substrain, REFMA) and the Tanzanian (KWA) strains. The high degree of segregation (95%) in hydrocarbons among the four strains investigated indicates that further analysis is needed to enable understanding of hydrocarbon variation in samples of An. gambiae especially from areas where these populations co-exist.

Selection of Trypanosoma cruzi clonal genotypes (clonet 20 and 39) isolated from Bolivian triatomines following subculture in liquid medium

Previous studies showed that two groups of Trypanosoma cruzi clonal genotypes named clonet 20 and clonet 39 were predominant in Triatoma infestans, the unique vector of Chagas disease in Bolivia. These groups of clones correspond to distinct genetic clusters. These clonets were detected in T. infestans and Rhodnius pictipes fecal samples before isolation and after culture by kDNA PCR (polymerase chain rreaction) and hybridization of the amplified products with clonet specific kDNA probes named 20 and 39 as previously reported. Forty eight T. infestans and three R. pictipes infected insects captured at random in different Bolivian departments were proceeded. As previously reported the direct identification of the two major clonets in fecal samples allowed the detection of abundant mixed infections: 41% in the original sample, however after culture, only 6% of mixed infections were detected. Among the 21 parasite stocks isolated from digestive tracts where mixed infections were initially detected (clonet 20 + 39) clonet 20 alone was detected in 81% of them. This result clearly showed that the culture step selected clonet 20 parasites over those belonging to clonet 39. The taxonomic status of the isolated stocks was also confirmed by isoenzyme typing, and correlation was observed between clustering topology and hybridization patterns with the probes 20 and 39.

Effect of CO2 and 1-octen-3-ol attractants for estimating species richness and the abundance of diurnal mosquitoes in the southeastern Atlantic forest, Brazil

Studies have shown that both carbon dioxide (CO2) and octenol (1-octen-3-ol) are effective attractants for mosquitoes. The objective of the present study was to evaluate the attractiveness of 1-octen-3-ol and CO2 for diurnal mosquitoes in the southeastern Atlantic forest. A Latin square experimental design was employed with four treatments: CDC-light trap (CDC-LT), CDC-LT and 1-octen-3-ol, CDC-LT and CO2 and CDC-LT with 1-octen-3-ol and CO2. Results demonstrated that both CDC-CO2 and CDC-CO2-1-octen-3-ol captured a greater number of mosquito species and specimens compared to CDC-1-octen-3-ol; CDC-LT was used as the control. Interestingly, Anopheles (Kerteszia) sp. was generally attracted to 1-octen-3-ol, whereas Aedes serratus was the most abundant species in all Latin square collections. This species was recently shown to be competent to transmit the yellow fever virus and may therefore play a role as a disease vector in rural areas of Brazil.

The impact of CO2 on collection of Aedes aegypti (Linnaeus) and Culex quinquefasciatus Say by BG-Sentinel(r) traps in Manaus, Brazil

Carbon dioxide (CO2) is an important component for activating and attracting host-seeking mosquitoes. The BG-Sentinel(r) trap is a well-established monitoring tool for capturing Culicidae, but CO2 role for the trap effectiveness has not been evaluated in highly urbanised areas. The objective was to evaluate the effectiveness of BG-Sentinel traps baited with and without CO2 for capturing urban mosquitoes. Fifteen areas were selected within the city of Manaus, Brazil, where four BG-Sentinels were operated for 24 h, two of them with CO2 and two without CO2. Captured Aedes aegypti females were dissected for the determination of their parity status. A significantly higher proportion of traps (from 32-79%) were positive for female Ae. aegypti when using the BG-Sentinel with CO2 (χ2 = 11.0271, p ≤ 0.001). Catches of female Culex spp were six times higher in CO2 traps (Mann-Whitney U test = 190.5; p = 0.001). Parity rates were similar for both traps. This study showed that CO2 has primarily an enhancing effect on the efficacy of BG-Sentinel for capturing Culex spp in Manaus. For Ae. aegypti, the positivity rate of the trap was increased, when CO2 was added.

Knockout confirmation for Hurries: rapid genotype identification of Trypanosoma cruzi transfectants by polymerase chain reaction directly from liquid culture

Gene knockout is a widely used approach to evaluate loss-of-function phenotypes and it can be facilitated by the incorporation of a DNA cassette having a drug-selectable marker. Confirmation of the correct knockout cassette insertion is an important step in gene removal validation and has generally been performed by polymerase chain reaction (PCR) assays following a time-consuming DNA extraction step. Here, we show a rapid procedure for the identification of Trypanosoma cruzi transfectants by PCR directly from liquid culture - without prior DNA extraction. This simple approach enabled us to generate PCR amplifications from different cultures varying from 106-108 cells/mL. We also show that it is possible to combine different primer pairs in a multiplex detection reaction and even to achieve knockout confirmation with an extremely simple interpretation of a real-time PCR result. Using the “culture PCR” approach, we show for the first time that we can assess different DNA sequence combinations by PCR directly from liquid culture, saving time in several tasks for T. cruzi genotype interrogation.

Using a single tablet daily to treat latent tuberculosis infection in Brazil: bioequivalence of two different isoniazid formulations (300 mg and 100 mg) demonstrated by a sensitive and rapid high-performance liquid chromatography-tandem mass spectrometry method in a randomised, crossover study

The recommended treatment for latent tuberculosis (TB) infection in adults is a daily dose of isoniazid (INH) 300 mg for six months. In Brazil, INH was formulated as 100 mg tablets. The treatment duration and the high pill burden compromised patient adherence to the treatment. The Brazilian National Programme for Tuberculosis requested a new 300 mg INH formulation. The aim of our study was to compare the bioavailability of the new INH 300 mg formulation and three 100 mg tablets of the reference formulation. We conducted a randomised, single dose, open label, two-phase crossover bioequivalence study in 28 healthy human volunteers. The 90% confidence interval for the INH maximum concentration of drug observed in plasma and area under the plasma concentration vs. time curve from time zero to the last measurable concentration “time t” was 89.61-115.92 and 94.82-119.44, respectively. The main limitation of our study was that neither adherence nor the safety profile of multiple doses was evaluated. To determine the level of INH in human plasma, we developed and validated a sensitive, simple and rapid high-performance liquid chromatography-tandem mass spectrometry method. Our results showed that the new formulation was bioequivalent to the 100 mg reference product. This finding supports the use of a single 300 mg tablet daily strategy to treat latent TB. This new formulation may increase patients’ adherence to the treatment and quality of life.

Biomassa microbiana e produção de C-CO2 e N mineral de um podzólico vermelho-escuro submetido a diferentes sistemas de manejo: Microbial biomass and C-CO2 and mineral nitrogen production in paleudult soil cultivated under different management systems

Os sistemas de manejo, com diferenças no revolvimento do solo e na composição dos resíduos vegetais, alteram as propriedades biológicas do solo, com reflexos na qualidade do solo e na produtividade das culturas. Com vistas em medir estas alterações nas propriedades biológicas do solo, a biomassa e a atividade microbiana foram avaliadas em um Podzólico Vermelho-Escuro, em Eldorado do Sul (RS), utilizando diferentes preparos (convencional, reduzido e plantio direto) e dois sistemas de sucessões de culturas (aveia preta + vica/milho + caupi e aveia/milho). As avaliações foram realizadas em quatro épocas, durante 12 meses, e em duas profundidades (0-5 e 5-15 cm). O carbono da biomassa microbiana foi analisado pelo método de fumigação-incubação, e a atividade microbiana, pela produção de C-CO2 e N mineral, após 60 dias de incubação. As diferenças na biomassa e na atividade microbiana, entre os sistemas de manejo, foram mais pronunciadas na camada de 0-5 cm. Nesta camada de solo, observaram-se os maiores valores de biomassa e de atividade nos preparos conservacionistas e no sistema aveia + vica/milho + caupi. Dentre as variáveis estudadas, a mineralização de N mostrou-se a mais sensível aos manejos, à profundidade e à época de amostragem.

Efeito de sistemas de preparo e de cultura na dinâmica da matéria orgânica e na mitigação das emissões de CO2

Neste estudo, ajustou-se um modelo exponencial de primeira ordem aos dados de carbono orgânico (CO) e N total (NT) do solo, do 5º e 9º ano de um experimento instalado em um Podzólico Vermelho-Escuro, em Eldorado do Sul (RS). Determinaram-se os parâmetros da dinâmica da matéria orgânica e simularam-se os estoques de CO e NT do solo em três sistemas de preparo (convencional-PC, reduzido-PR e plantio direto-PD) e três sistemas de cultura (aveia/milho-A/M, aveia + ervilhaca/milho-A + E/M e aveia + ervilhaca/milho + caupi A + E/M + C). A taxa de decomposição da matéria orgânica do solo diminuiu de 0,054 ano-1, no sistema com lavração e gradagem (PC), para 0,039 ano-1, no solo escarificado (PR), e 0,029 ano-1, no solo não revolvido (PD). Estimou-se que os estoques de CO e NT do solo em 1990 (30,78 Mg ha-1 e 2.200 kg ha-1) diminuirão para 16,11 Mg ha-1 e 1.396 kg ha-1 na combinação PC - A/M. Por sua vez, no sistema PD - A + E/M + C, os estoques de CO e NT do solo em 1990 (32,52 Mg ha-1 e 2.690 Mg ha-1) tenderão a aumentar a estoques estáveis de 54,83 Mg ha-1 e 7.966 kg ha-1, respectivamente. Os sistemas de manejo sem revolvimento do solo e alto aporte de resíduos apresentaram efeito positivo na mitigação das emissões de CO2. Enquanto o sistema PC-A/M apresentou um efluxo líquido (emitido pelo solo > fixado pelas culturas por fotossíntese) de 2,08 Mg CO2 ha-1 para atmosfera no ano de 1994, o sistema PD - A + E/M + C mostrou um influxo líquido (emitido pelo solo < fixado pelas culturas) de CO2 de 1,79 Mg ha-1 ano-1. Atingidos os estoques estáveis de CO no solo estimados pelo modelo, o solo no sistema PD - A + E/M + C terá seqüestrado aproximadamente 142 Mg CO2 ha-1, em comparação ao sistema PC-A/M.

Rotação de culturas no sistema plantio direto em Tibagi (PR): II - Emissões de CO2 e N2O

A atividade agrícola pode alterar a quantidade e qualidade da matéria orgânica do solo (MOS), resultando em emissões de dióxido de carbono (CO2) e óxido nitroso (N2O) do solo para a atmosfera. O sistema plantio direto (SPD) com a utilização de leguminosas em sistemas de rotação é uma estratégia que deve ser considerada tanto para o aumento da quantidade de MOS como para seu efeito na redução das emissões dos gases de efeito estufa. Com o objetivo de determinar os fluxos de gases do efeito estufa (CO2 e N2O) do solo, um experimento foi instalado em Tibagi (PR), em um Latossolo Vermelho distroférrico textura argilosa. Os tratamentos, dispostos em faixas não casualizadas com parcelas subdivididas, foram: sistema plantio direto por 12 anos com sucessões milho/trigo e soja/trigo (PD12 M/T e PD12 S/T, respectivamente) e por 22 anos (PD22 M/T e PD22 S/T, respectivamente). As emissões de CO2 do solo foram aproximadamente 20 % mais elevadas no PD22 em relação ao PD12. As emissões de CO2 apresentaram correlação significativa (R² = 0,85; p < 0,05) com a temperatura do solo, com emissões médias 40 % menores, registradas nos meses com temperaturas mais baixas. As emissões mais elevadas de N2O foram observadas após a colheita das culturas de verão, sobretudo na sucessão milho/trigo, em relação à sucessão soja/trigo. As emissões de N2O foram aproximadamente 25 % maiores após aplicação do fertilizante nitrogenado na cultura do trigo nas duas sucessões e apresentaram correlação significativa (R² = 0,88; p < 0,01) com o grau de saturação de água no solo (Sr %).

CO2, CH4 and N2O fluxes in an Ultisol treated with sewage sludge and cultivated with castor bean

Organic residue application into soil alter the emission of gases to atmosphere and CO2, CH4, N2O may contribute to increase the greenhouse effect. This experiment was carried out in a restoration area on a dystrophic Ultisol (PVAd) to quantify greenhouse gas (GHG) emissions from soil under castor bean cultivation, treated with sewage sludge (SS) or mineral fertilizer. The following treatments were tested: control without N; FertMin = mineral fertilizer; SS5 = 5 t ha-1 SS (37.5 kg ha-1 N); SS10 = 10 t ha-1 SS (75 kg ha-1 N); and SS20 = 20 t ha-1 SS (150 kg ha-1 N). The amount of sludge was based on the recommended rate of N for castor bean (75 kg ha-1), the N level of SS and the mineralization fraction of N from SS. Soil gas emission was measured for 21 days. Sewage sludge and mineral fertilizers altered the CO2, CH4 and N2O fluxes. Soil moisture had no effect on GHG emissions and the gas fluxes was statistically equivalent after the application of FertMin and of 5 t ha-1 SS. The application of the entire crop N requirement in the form of SS practically doubled the Global Warming Potential (GWP) and the C equivalent emissions in comparison with FertMin treatments.

Long-term C-CO2 emissions and carbon crop residue mineralization in an oxisol under different tillage and crop rotation systems

Soil C-CO2 emissions are sensitive indicators of management system impacts on soil organic matter (SOM). The main soil C-CO2 sources at the soil-plant interface are the decomposition of crop residues, SOM turnover, and respiration of roots and soil biota. The objectives of this study were to evaluate the impacts of tillage and cropping systems on long-term soil C-CO2 emissions and their relationship with carbon (C) mineralization of crop residues. A long-term experiment was conducted in a Red Oxisol in Cruz Alta, RS, Brazil, with subtropical climate Cfa (Köppen classification), mean annual precipitation of 1,774 mm and mean annual temperature of 19.2 ºC. Treatments consisted of two tillage systems: (a) conventional tillage (CT) and (b) no tillage (NT) in combination with three cropping systems: (a) R0- monoculture system (soybean/wheat), (b) R1- winter crop rotation (soybean/wheat/soybean/black oat), and (c) R2- intensive crop rotation (soybean/ black oat/soybean/black oat + common vetch/maize/oilseed radish/wheat). The soil C-CO2 efflux was measured every 14 days for two years (48 measurements), by trapping the CO2 in an alkaline solution. The soil gravimetric moisture in the 0-0.05 m layer was determined concomitantly with the C-CO2 efflux measurements. The crop residue C mineralization was evaluated with the mesh-bag method, with sampling 14, 28, 56, 84, 112, and 140 days after the beginning of the evaluation period for C measurements. Four C conservation indexes were used to assess the relation between C-CO2 efflux and soil C stock and its compartments. The crop residue C mineralization fit an exponential model in time. For black oat, wheat and maize residues, C mineralization was higher in CT than NT, while for soybean it was similar. Soil moisture was higher in NT than CT, mainly in the second year of evaluation. There was no difference in tillage systems for annual average C-CO2 emissions, but in some individual evaluations, differences between tillage systems were noticed for C-CO2 evolution. Soil C-CO2 effluxes followed a bi-modal pattern, with peaks in October/November and February/March. The highest emission was recorded in the summer and the lowest in the winter. The C-CO2 effluxes were weakly correlated to air temperature and not correlated to soil moisture. Based on the soil C conservation indexes investigated, NT associated to intensive crop rotation was more C conserving than CT with monoculture.

Soil CO2 flux: a method comparison of closed static chambers in a sugarcane field

A large variety of techniques have been used to measure soil CO2 released from the soil surface, and much of the variability observed between locations must be attributed to the different methods used by the investigators. Therefore, a minimum protocol of measurement procedures should be established. The objectives of this study were (a) to compare different absorption areas, concentrations and volumes of the alkali trapping solution used in closed static chambers (CSC), and (b) to compare both, the optimized alkali trapping solution and the soda-lime trapping using CSC to measure soil respiration in sugarcane areas. Three CO2 absorption areas were evaluated (7; 15 and 20 % of the soil emission area or chamber); two volumes of NaOH (40 and 80 mL) at three concentrations (0.1, 0.25 and 0.5 mol L-1). Three different types of alkaline traps were tested: (a), 80 mL of 0.5 mol L-1 NaOH in glass containers, absorption area 15 % (V0.5); (b) 40 mL of 2 mol L-1 NaOH retained in a sponge, absorption area 80 % (S2) and (c) 40 g soda lime, absorption area 15 % (SL). NaOH concentrations of 0.5 mol L-1 or lower underestimated the soil CO2-C flux or CO2 flux. The lower limit of the alkali trap absorption area should be a minimum of 20 % of the area covered by the chamber. The 2 mol L-1 NaOH solution trap (S2) was the most efficient (highest accuracy and highest CO2 fluxes) in measuring soil respiration.

Uncertainties in the prediction of spatial variability of soil CO2 emissions and related properties

The soil CO2 emission has high spatial variability because it depends strongly on soil properties. The purpose of this study was to (i) characterize the spatial variability of soil respiration and related properties, (ii) evaluate the accuracy of results of the ordinary kriging method and sequential Gaussian simulation, and (iii) evaluate the uncertainty in predicting the spatial variability of soil CO2 emission and other properties using sequential Gaussian simulations. The study was conducted in a sugarcane area, using a regular sampling grid with 141 points, where soil CO2 emission, soil temperature, air-filled pore space, soil organic matter and soil bulk density were evaluated. All variables showed spatial dependence structure. The soil CO2 emission was positively correlated with organic matter (r = 0.25, p < 0.05) and air-filled pore space (r = 0.27, p < 0.01) and negatively with soil bulk density (r = -0.41, p < 0.01). However, when the estimated spatial values were considered, the air-filled pore space was the variable mainly responsible for the spatial characteristics of soil respiration, with a correlation of 0.26 (p < 0.01). For all variables, individual simulations represented the cumulative distribution functions and variograms better than ordinary kriging and E-type estimates. The greatest uncertainties in predicting soil CO2 emission were associated with areas with the highest estimated values, which produced estimates from 0.18 to 1.85 t CO2 ha-1, according to the different scenarios considered. The knowledge of the uncertainties generated by the different scenarios can be used in inventories of greenhouse gases, to provide conservative estimates of the potential emission of these gases.

Efeito do preparo do solo e resíduo da colheita de cana-de-açúcar sobre a emissão de CO2

O solo é um dos principais compartimentos de carbono no ecossistema terrestre, capaz de armazenar quantidades expressivas desse elemento e, portanto, a compreensão dos fatores que contribuem para as perdas de CO2 em solos agrícolas é fundamental para determinar estratégias de redução das emissões desse gás e ajudar a mitigar o efeito estufa. O objetivo deste estudo foi investigar o efeito do preparo do solo e da deposição de resíduos da cultura da cana-de-açúcar na emissão de CO2, temperatura e umidade do solo, durante a reforma do canavial, ao longo de um período de 15 dias. Os manejos avaliados foram: sem preparo do solo e mantendo os resíduos da colheita sobre a superfície do solo (SPCR); sem preparo do solo e sem resíduo (SPSR) e com preparo do solo e sem resíduo (CPSR). A menor média de emissão de CO2 do solo (FCO2) foi observada no manejo SPCR (2,16 µmol m-2 s-1), quando comparado aos manejos SPSR (2,90 µmol m-2 s-1) e CPSR (3,22 µmol m-2 s-1), indicando que as maiores umidades e menores variações da temperatura do solo, observadas em SPCR, foram os fatores responsáveis por tal diminuição. Durante o período de estudo, a menor média diária da FCO2 foi registrada em SPCR (1,28 µmol m-2 s-1) e a maior em CPSR (6,08 µmol m-2 s-1), após a ocorrência de chuvas. A menor perda de C-CO2 do solo foi observada no manejo SPCR (367 kg ha-1 de C-CO2), diferindo significativamente (p<0,05) dos manejos: SPSR (502 kg ha-1 de C-CO2) e CPSR (535 kg ha-1 de C-CO2). A umidade do solo foi a variável que apresentou valores mais diferenciados entre os manejos, sendo positivamente correlacionada (r = 0,55; p<0,05) com as variações temporais da emissão de CO2 nos manejos SPCR e CPSR. Em adição, a temperatura do solo diferiu (p<0,05) somente no manejo SPCR (24 ºC), quando comparada aos manejos SPSR (26 ºC) e CPSR (26,5 ºC), sugerindo que, para as condições deste estudo, o resíduo da cana-de-açúcar retido sobre a superfície propiciou uma temperatura do solo, em média, 2 ºC mais amena.