Evaluation of rapid alternative methods for drug susceptibility testing in clinical isolates of Mycobacterium tuberculosis

A study was carried out to compare the performance of a commercial method (MGIT) and four inexpensive drug susceptibility methods: nitrate reductase assay (NRA), microscopic observation drug susceptibility (MODS) assay, MTT test, and broth microdilution method (BMM). A total of 64 clinical isolates of Mycobacterium tuberculosis were studied. The Lowenstein-Jensen proportion method (PM) was used as gold standard. MGIT, NRA, MODS, and MTT results were available on an average of less than 10 days, whereas BMM results could be reported in about 20 days. Most of the evaluated tests showed excellent performance for isoniazid and rifampicin, with sensitivity and specificity values > 90%. With most of the assays, sensitivity for ethambutol was low (62-87%) whereas for streptomycin, sensitivity values ranged from 84 to 100%; NRA-discrepancies were associated with cultures with a low proportion of EMB-resistant organisms while most discrepancies with quantitative tests (MMT and BMM) were seen with isolates whose minimal inhibitory concentrations fell close the cutoff. MGIT is reliable but still expensive. NRA is the most inexpensive and easiest method to perform without changing the organization of the routine PM laboratory performance. While MODS, MTT, and BMM, have the disadvantage from the point of view of biosafety, they offer the possibility of detecting partial resistant strains. This study shows a very good level of agreement of the four low-cost methods compared to the PM for rapid detection of isoniazid, rifampicin and streptomycin resistance (Kappa values > 0.8); more standardization is needed for ethambutol.

Evaluation of buffy coat 16S rRNA PCR, buffy coat culture and whole blood PCR for detection of bacteraemia

The use of Gram type-specific PCR on buffy coat from clinical specimens for the detection of bacteraemia was evaluated for the first time using whole blood culture as the gold standard. In addition, the established buffy coat culture and whole blood PCR were also compared. Gram-positive bacteria belonging to six species and Gram-negative bacteria from 10 species were isolated and identified by culture and detected using broad-range 16S rDNA primers and Gram-specific primers. Data from the three methods all conferred very high sensitivity, specificity, positive and negative predictive values when compared to whole blood culture. The Kappa coefficients of agreement were 0.9819 (buffy coat PCR), 0.9458 (whole blood PCR) and 1.0 (buffy coat culture), which establishes their validity as alternative methods to routine blood culture in detecting bacteraemia. In addition, results showed that there was a direct correlation of WBC counts greater than 12,000 cells per mm³ to the occurrence of bacteraemia as detected by the four methods (p < 0.05).

Is imidacloprid an effective alternative for controlling pyrethroid-resistant populations of Triatoma infestans (Hemiptera: Reduviidae) in the Gran Chaco ecoregion?

The prevention of Chagas disease is based primarily on the chemical control of Triatoma infestans (Klug) using pyrethroid insecticides. However, high resistance levels, correlated with control failures, have been detected in Argentina and Bolivia. A previous study at our laboratory found that imidacloprid could serve as an alternative to pyrethroid insecticides. We studied the delayed toxicity of imidacloprid and the influence of the blood feeding condition of the insect on the toxicity of this insecticide; we also studied the effectiveness of various commercial imidacloprid formulations against a pyrethroid-resistant T. infestans population from the Gran Chaco ecoregion. Variations in the toxic effects of imidacloprid were not observed up to 72 h after exposure and were not found to depend on the blood feeding condition of susceptible and resistant individuals. Of the three different studied formulations of imidacloprid on glass and filter paper, only the spot-on formulation was effective. This formulation was applied to pigeons at doses of 1, 5, 20 and 40 mg/bird. The nymphs that fed on pigeons treated with 20 mg or 40 mg of the formulation showed a higher mortality rate than the control group one day and seven days post-treatment (p < 0.01). A spot-on formulation of imidacloprid was effective against pyrethroid-resistant T. infestans populations at the laboratory level.

Development of a control alternative for the citrus fruit borer, Ecdytolopha aurantiana (Lepidoptera, Tortricidae): from basic research to the grower

All research steps, developed from 1995 to 2000, to synthesize the sex pheromone of Ecdytolopha aurantiana (Lima, 1927) are described, in order to monitoring this pest that causes losses in the order of 50 million dollars per year to citriculture in the State of São Paulo. The basic researches conducted are described, including the development of an artificial diet for the insect, the study of its temperature and humidity requirements, behavioral studies, and synthesis of the male-attracting substance up to the formulation and distribution of the pheromone to the grower, by means of its commercialization. It is a case of success, at a cost of 50 thousand dollars, involving inter- and multidisciplinary researches, which can be adopted to other insect pests in the country.

Evaluation of Sikora instead of SMP buffer to estimate the potential acidity of brazilian soils

Despite the efficiency of the Shoemaker, McLean, Pratt (SMP) buffer method in estimating soil acidity, the presence of p-nitrophenol and potassium chromate in the solution, both hazardous substances, has caused increasing environmental concerns. The purpose of this study was to test Sikora method (Sikora, 2006) as an alternative to the adapted SMP buffer method, generally used to estimate potential acidity of Southern Brazilian soils. For the test, 21 soils in the South and Cerrado regions of Brazil were sampled. (1) The potential acidity values of these soils range from 35.95 to 4.02 cmol c kg-1 of soil, reflecting a wide acidity variation. The Sikora buffer does not mimic the adapted SMP buffer used in Southern Brazil, since the former has a low ability to distinguish soils with different acidity from each other, probably due to the higher buffer capacity than of the adapted SMP solution.

An improved method for measuring soil microbial activity by gas phase flow injection analysis

The rate of carbon dioxide production is commonly used as a measure of microbial activity in the soil. The traditional method of CO2 determination involves trapping CO2 in an alkali solution and then determining CO2 concentration indirectly by titration of the remaining alkali in the solution. This method is still commonly employed in laboratories throughout the world due to its relative simplicity and the fact that it does not require expensive, specific equipment. However, there are several drawbacks: the method is time-consuming, requires large amounts of chemicals and the consistency of results depends on the operator's skills. With this in mind, an improved method was developed to analyze CO2 captured in alkali traps, which is cheap and relatively simple, with a substantially shorter sample handling time and reproducibility equivalent to the traditional titration method. A comparison of the concentration values determined by gas phase flow injection analysis (GPFIA) and titration showed no significant difference (p > 0.05), but GPFIA has the advantage that only a tenth of the sample volume of the titration method is required. The GPFIA system does not require the purchase of new, costly equipment but the device was constructed from items commonly found in laboratories, with suggestions for alternative configurations for other detection units. Furthermore, GPFIA for CO2 analysis can be equally applied to samples obtained from either the headspace of microcosms or from a sampling chamber that allows CO2 to be released from alkali trapping solutions. The optimised GPFIA method was applied to analyse CO2 released from degrading hydrocarbons from a site contaminated by diesel spillage.

Year-round poultry litter decomposition and N, P, K and Ca release

Poultry litter is an important nutrient source in agriculture, although little information is available regarding its decomposition rate and nutrient release. To evaluate these processes, poultry litter (PL) was applied to the soil to supply 100, 200 and 300 kg ha-1 N contained in 4,953, 9,907 and 14,860 kg ha-1 PL, respectively. The litter bag technique was used to monitor the process of decomposition and nutrient release from the litter. These bags were left on the soil surface and collected periodically (after 15, 30, 60, 90, 120, 150, 180, 210, 240, 270, 300, 330, and 365 days). The dry matter (DM) loss was highest (35 %) after the first 30 days of field incubation. The highest nutrient release occurred in the first 60 days on the field, when 40, 34, 91, and 39 %, respectively, of N, P, K, and Ca of the initial PL dry matter (4,860 kg ha-1) was already released to the soil. In absolute terms, these percentages represent 40, 23, 134, and 69 kg ha-1 of N, P, K, and Ca and these values doubled and tripled as the PL fertilization rates increased to 9,907 and 14,860 kg ha-1, respectively. After one year of field incubation, the residual contents in the litter were 27, 15, 18 and 30 % of the initial DM , and N, P and Ca, respectively. The release rate of K was the fastest and 91 % of the K had been released from the PL after 30 days of field incubation.

EFFECT OF ALTERNATIVE MULTINUTRIENT SOURCES ON SOIL CHEMICAL PROPERTIES

The current high price of potassium chloride and the dependence of Brazil on imported materials to supply the domestic demand call for studies evaluating the efficiency of alternative sources of nutrients. The aim of this work was to evaluate the effect of silicate rock powder and a manganese mining by-product, and secondary materials originated from these two materials, on soil chemical properties and on brachiaria production. This greenhouse experiment was conducted in pots with 5 kg of soil (Latossolo Vermelho-Amarelo distrófico - Oxisol). The alternative nutrient sources were: verdete, verdete treated with NH4OH, phonolite, ultramafic rock, mining waste and the proportion of 75 % of these K fertilizers and 25 % lime. Mixtures containing 25 % of lime were heated at 800 ºC for 1 h. These sources were applied at rates of 0, 150, 300, 450 and 600 kg ha-1 K2O, and incubated for 45 days. The mixtures of heated silicate rocks with lime promoted higher increases in soil pH in decreasing order: ultramafic rock>verdete>phonolite>mining waste. Applying the mining waste-lime mixture increased soil exchangeable K, and available P when ultramafic rock was incorporated. When ultramafic rock was applied, the release of Ca2+ increased significantly. Mining subproduct released the highest amount of Zn2+ and Mn2+ to the soil. The application of alternative sources of K, with variable chemical composition, altered the nutrient availability and soil chemical properties, improving mainly plant development and K plant uptake, and are important nutrient sources.

LEAF TOTAL NITROGEN CONCENTRATION AS AN INDICATOR OF NITROGEN STATUS FOR PLANTLETS AND YOUNG PLANTS OF EUCALYPTUS CLONES

The use of leaf total nitrogen concentration as an indicator for nutritional diagnosis has some limitations. The objective of this study was to determine the reliability of total N concentration as an indicator of N status for eucalyptus clones, and to compare it with alternative indicators. A greenhouse experiment was carried out in a randomized complete block design in a 2 × 6 factorial arrangement with plantlets of two eucalyptus clones (140 days old) and six levels of N in the nutrient solution. In addition, a field experiment was carried out in a completely randomized design in a 2 × 2 × 2 × 3 factorial arrangement, consisting of two seasons, two regions, two young clones (approximately two years old), and three positions of crown leaf sampling. The field areas (regions) had contrasting soil physical and chemical properties, and their soil contents for total N, NH+4-N, and NO−3-N were determined in five soil layers, up to a depth of 1.0 m. We evaluated the following indicators of plant N status in roots and leaves: contents of total N, NH+4-N, NO−3-N, and chlorophyll; N/P ratio; and chlorophyll meter readings on the leaves. Ammonium (root) and NO−3-N (root and leaf) efficiently predicted N requirements for eucalyptus plantlets in the greenhouse. Similarly, leaf N/P, chlorophyll values, and chlorophyll meter readings provided good results in the greenhouse. However, leaf N/P did not reflect the soil N status, and the use of the chlorophyll meter could not be generalized for different genotypes. Leaf total N concentration is not an ideal indicator, but it and the chlorophyll levels best represent the soil N status for young eucalyptus clones under field conditions.

Fruit quality of 'Tahiti' lime (Citrus latifolia Tanak) grafted on twelve different rootstocks

In the present work, the effect of twelve rootstocks on fruit quality of the nucellar clone IAC-5 of 'Tahiti' lime, (Citrus latifolia Tanaka) and the influence of fruit position on tree in fruit quality was evaluated in the Citrus Experimental Station of Bebedouro (EECB), located in the Bebedouro county, state of São Paulo, Brazil. A 8.0 x 5.0m planting frame was utilized. The evaluated rootstocks were: 'Carrizo' citrange (C. sinensis (L.) Osbeck x Poncirus trifoliata (L.) Raf.); the hybrids 'Rangpur' lime x 'Swingle' citrumello (C. limonia Osbeck x P. trifoliata Raf) and 'Changsha' x 'English Small'(C. sunki Hort. ex Tan. x P. trifoliata Raf.); the mandarins 'Sun Chu Sha Kat' (C. reticulata Blanco) and 'Sunki' (C. sunki Hort. ex Tanaka); the 'Rangpur' limes 'Cravo Limeira' and ' Cravo FCAV' (C. limonia Osbeck); the 'Swingle' citrumello (P. trifoliata Raf. x C. paradisi Macf.); the 'Orlando' tangelo (C. reticulata Blanco x C. paradisi Macf.) and the trifoliates cvs. 'Rubidoux', 'FCAV' and 'Flying Dragon' (P. trifoliata Raf.). The experiment was arranged in a randomized block design, with twelve treatments, six replicates and one plant per plot. The rootstocks induced differences in fruit quality; however, all the evaluated quality characteristics were within the values considered as normal and acceptable for the variety, constituting good alternative rootstocks for the 'Rangpur' lime. Additionally, the fruit position in the plant (northeastern or southwestern) had a significant influence on the external fruit color regardless of the rootstock.

Sílica quimicamente modificada com os grupos p-anisidina, p-fenitidina e p-fenilenodiamina usada como adsorvente para Pb2+, Cu2+, Cd2+ e Ni2+ em soluções aquosa e etanólica

Silica gel was chemically modified with the aromatic amines p-anisidine, p-phenytidine and p-phenylenediamine, using grafting reactions. The resulting modified silicas were characterized by infrared spectroscopy and N2 adsorption/desorption isotherms. The organic groups were covalently immobilized in a monolayer form. These modified silicas were investigated as adsorbents for Pb2+, Cu2+, Cd2+ and Ni2+ in aqueous and ethanol solutions. In a general way, the adsorption capacity values for all adsorbents presented the following sequence: Pb2+ >> Cu2+ @Cd2+ @ Ni2+. Adsorption studies for all adsorbents, in competitive medium, showed better selectivity for Cu2+ and Pb2+ in aqueous medium and for Pb2+ in ethanol solution. Desorption studies were carried out using HCl and HNO3 as eluents.

Analysis of the relationship intensity, duration, frequency of disaggregated daily rainfall in southern Rio Grande do Sul, Brazil

The intensity, duration, and frequency relationship (IDF) of rainfall occurrence may be done through continuous records of pluviographs or daily pluviometer values . The objective of this study was to estimate the intensity-duration-frequency relationships of precipitation, using the method of daily rainfall disaggregation, at weather stations located to the southern half of the state of Rio Grande do Sul; comparing them with those obtained by rain gauge records, in places considered homogeneous from the meteorological point of view. The IDF equation parameters were estimated from daily rainfall disaggregation data, using the method of nonlinear optimization. To validate the equations confidence indices and efficiency and the "t" Student test, among maximum intensity values obtained from the disaggregated daily rainfall durations of 10; 30; 60 min and 6; 12 and 24 h and those extracted from existing IDF equations. For all studied stations and return periods, the trust index values were regarded as "optimal", i.e., greater than 0.85. The maximal intensity of rainfall obtained by daily rainfall disaggregation have similarity with those obtained by relations IDF standards. Thus, the method constitutes a feasible alternative in obtaining the IDF relationships.

Cloroplast pigments as indicators of lead stress

Plants respond to environmental adversities, becoming an indicator for assessing the environment quality. In this aspect, chlorophyll contents as well, carotenoids are used as a reliable indicator to associate environmental quality and pollution, mainly regarding the toxicity of heavy metals in higher plants. So, we aimed to evaluate the content of chlorophyll a, b, and total chlorophylls and carotenoids in plants vetiver [Vetiveria zizanioides (L.) Nash], maize (Zea mays L.) cv. AG 1051, sunflower (Helianthus annuus L.) cv. BRS 122/V-2000, and castor beans (Ricinus communis L. ) cv . Northeastern BRS grown in contaminated soil with lead, with and without correction of soil pH, so they were used as indicators of metal stress by the soil. From the biochemical point of view, the correction of soil pH values caused chlorophyll a, b and total statistically higher for vetiver species and castor beans in the analyzed periods, except for the analysis performed 60 days after transplanting where only the species vetiver benefited from the correction of soil pH on the content of chlorophyll b and total. On the other hand plants without correction of soil pH showed a decrease of all chlorophyll levels. In addition, the largest increase in the synthesis of carotenoids, indicated that under stress the plants have developed alternative routes of dissipation of energy in order to avoid problems of photo-inhibition and photo-oxidation.

Test for alternative indicator of soybean sowing depth

Among studies focused on increasing soybean grain yield, the ones related to sowing process are the most significant. Considering that soybean has an epigeal emergence, it becomes difficult to hint at the length covered by hypocotyl up to soil surface, or the actual planting depth. This study aimed to find an indicator that allows the identification of an ideal soybean planting depth. For this purpose, two complementary assays has been carried out in a greenhouse. The first aimed to identify structures that could be indicators of seed planting depth, on a medium-textured soil from Campos Gerais region, in the state of Paraná, Brazil. Spring NK 8350 cultivar seeds were sown at five theoretical depths (1, 2, 3, 4 and 5 cm). As seedlings emerged, the “differentiation zone” and the “root curve” depths were measured. The second assay was the validation of the suggested indicators in assay 1 from two soils, one medium-textured and one clay-textured. For this assay, it was used BRS 232. Both the methodologies showed high correlation with the theoretical planting depth. Although their correlation coefficient values were close, the differentiation zone appeared to be the most efficient reference with less planting depth overestimation.

Application of artificial neural networks as an alternative to volumetric water balance in drip irrigation management in watermelon crop

Precision irrigation seeks to establish strategies which achieve an efficient ratio between the volume of water used (reduction in input) and the productivity obtained (increase in production). There are several studies in the literature on strategies for achieving this efficiency, such as those dealing with the method of volumetric water balance (VWB). However, it is also of great practical and economic interest to set up versatile implementations of irrigation strategies that: (i) maintain the performance obtained with other implementations, (ii) rely on few computational resources, (iii) adapt well to field conditions, and (iv) allow easy modification of the irrigation strategy. In this study, such characteristics are achieved when using an Artificial Neural Network (ANN) to determine the period of irrigation for a watermelon crop in the Irrigation Perimeter of the Lower Acaraú, in the state of Ceará, Brazil. The Volumetric Water Balance was taken as the standard for comparing the management carried out with the proposed implementation of ANN. The statistical analysis demonstrates the effectiveness of the proposed management, which is able to replace VWB as a strategy in automation.