Determining aflatoxins B1, B2, G1 and G2 in maize using florisil clean up with thin layer chromatography and visual and densitometric quantification

A method for determining aflatoxins B1 (AFB1), B2 (AFB2),G1 (AFG1) andG2 (AFG2) in maize with florisil clean up was optimised aiming at one-dimensional thin layer chromatography (TLC) analysis with visual and densitometric quantification. Aflatoxins were extracted with chloroform: water (30:1, v/v), purified through florisil cartridges, separated on TLC plate, detected and quantified by visual and densitometric analysis. The in-house method performance characteristics were determined by using spiked, naturally contaminated maize samples, and certified reference material. The mean recoveries for aflatoxins were 94.2, 81.9, 93.5 and 97.3% in the range of 1.0 to 242 µg/kg for AFB1, 0.3 to 85mg/kg for AFB2, 0.6 to 148mg/kg for AFG1 and 0.6 to 140mg/kg for AFG2, respectively. The correlation values between visual and densitometric analysis for spiked samples were higher than 0.99 for AFB1, AFB2, AFG1 and 0.98 for AFG2. The mean relative standard deviations (RSD) for spiked samples were 16.2, 20.6, 12.8 and 16.9% for AFB1, AFB2, AFG1 and AFG2, respectively. The RSD of the method for naturally contaminated sample (n = 5) was 16.8% for AFB1 and 27.2% for AFB2. The limits of detection of the method (LD) were 0.2, 0.1, 0.1 and 0.1mg/kg and the limits of quantification (LQ) were 1.0, 0.3, 0.6 and 0.6mg/kg for AFB1, AFB2, AFG1 and AFG2, respectively.

Five generations of peace operations: from the "thin blue line" to "painting a country blue"

This article combines both international practice and analytical contributions into a systematic and synthetic presentation of the evolution of peace operations from their modern inception in 1948 to the present. It seeks to serve a didactic purpose in proposing a basic structure for Brazilian scholars' burgeoning debate on peace operations and intervention, rather than a definitive characterization of blue helmet practice. Peace operations' progression is traced through five analytical "generations," each adding a crucial factor distinguishing it from its predecessors. Each generation is placed in relation to changes in the nature of conflict and in the interpretation of the foundational principles of peace operations, and links to broader theoretical issues in International Relations are made explicit at each stage.

Screening the mutagenic activities of coomonly used antiparasite drugs by the Simultest, a simplified Salmonella/microsome plate incorporation assay

The mutagenic activities of 16 anti-parasite drugs were screened by the Simultest in both qualitative (spot test) and quantitative (plate incorporation) assays with a Salmonella typhimurium pool composed by the indicator strains TA97, TA 98, TA100 and TA102. Four anti Chagas' disease drugs (nifurtimox, benznidazole, CL 64,855, and MK 436) and two anti-amebae drugs (metronidazole and tinidazole) gave positive results in qualitative tests and incorporation of rat liver microsomes did not alter the results. Comparative dose response curves of the mutagenic activities of CL 64,855, metronidazole and benznidazole obtained by the simultest and by individual Salmonella indicator strains demonstrated that both approachs have similar sensitivities. The results corroborate the validity of the Simultest, as a simplified, fast and economic version of the Ames test in preliminary screening of potential mutagenic drugs.

Studies on prevalence of Strongyloides infection in Holambra and Maceió, Brazil, by the agar plate faecal culture method

Prevalence of Strongyloides stercoralis infection in three areas of Brazil was surveyed by a recently developed faecal culture method (an agar plate culture). The Strongyloides infection was confirmed in 11.3% of 432 subjects examined. The diagnostic efficacy of the agar plate culture was as high as 93.9% compared to only 28.5% and 26.5% by the Harada-Mori filter paper culture and faecal concentration methods, when faecal samples were examined simultaneously by these three methods. Among the 49 positive samples, about 60% were confirmed to be positive only by the agar plate culture. These results indicate that the agar plate culture is a sensitive new tool for the correct diagnosis of chronic Strongyloides infection.

APPLICATION OF 6-NITROCOUMARIN AS A SUBSTRATE FOR THE FLUORESCENT DETECTION OF NITROREDUCTASE ACTIVITY IN Sporothrix schenckii

Introduction Sporothrix schenckii is a thermal dimorphic pathogenic fungus causing a subcutaneous mycosis, sporotrichosis. Nitrocoumarin represents a fluorogenic substrate class where the microbial nitroreductase activity produces several derivatives, already used in several other enzyme assays. The objective of this study was the analysis of 6-nitrocoumarin (6-NC) as a substrate to study the nitroreductase activity in Sporothrix schenckii. Methods Thirty-five samples of S. schenckii were cultivated for seven, 14 and 21 days at 35 °C in a microculture containing 6-nitrocoumarin or 6-aminocoumarin (6-AC) dissolved in dimethyl sulfoxide or dimethyl sulfoxide as a negative control, for posterior examination under an epifluorescence microscope. The organic layer of the seven, 14 and 21-day cultures was analyzed by means of direct illumination with 365 nm UV light and by means of elution on G silica gel plate with hexane:ethyl acetate 1:4 unveiled with UV light. Results All of the strains showed the presence of 6-AC (yellow fluorescence) and 6-hydroxylaminocoumarin (blue fluorescence) in thin layer chromatography, which explains the green fluorescence observed in the fungus structure. Conclusion The nitroreductase activity is widely distributed in the S. schenckii complex and 6-NC is a fluorogenic substrate of easy access and applicability for the nitroreductase activity detection.

Physa Marmorata Guilding, 1828 (Pulmonata: Physidae)

A description of Physa marmorata Guilding, 1828, based on material collected at its type-locality, the Caribbean island of Saint Vincent, is presented. The shell is thin, horn-colored, surface very glossy, diaphanous. Spire acute, elevated; protoconch distinct, rounded-conical, reddish-brown; five not shouldered, broadly convex whorls with subobsolete spiral lines and thin growth lines. Aperture elongated, 1.4-2.0 times as long as the remaining shell length, narrow obovate-lunate; upper half acute-angled,lower half oval,narrowly rounded at the base, outer lip sharp, inner lip completely closing the umbilical region; a very distinct callus on the parietal wall; columellar lip with a low ridge gradually merging into the callus. ratios: shell width/shell length = 0.44 - 0.52 (mean 0.47); spire length /shell lenght = 0.33-0.41 (mean 0.39); aperture length/shell lenght = 0.59-0.67 (mean 0.62). Oral lappets laterally mucronate, foot spatulate with deeply pigmented acuminate tail. Mantle reflection with 6-10 short triangular dentations covering nearly half the right surface of the body whorl, and 4-6 covering a part of the ventral wall. Body surface with tiny dots of greenish-yellow pigment besides melanin. Renal tube tightly folded in toa zigzag course. Ovotestis diverticula acinous, laterally pressed against each other around a collecting canal. Ovispermiduct with well-developed seminal vesicle. oviduct highly convoluted, merging into a less convoluted nidamental gland which narrows to a funnel-shaped uterus and a short vagina. Spermathecal body oblong, more or less constricted in the middle and somewhat curved; spermathecal duct uniformly narrow, a little longer than be body. About 20 prostatic diverticula, simple, bifurcate or divided into a few short branches, distalmost ones assembled into a cluster. Penis long, nearly uniformly narrow; penial canal with lateral opening about the junction of its middle and lower thirds. Penial sheath with a bulbous terminal expasion the tip of which isinserted into the caudal end of the prepuce. Prepuce shouldered, much wider than the narrow portion of the penial sheath. Penial sheath/prepuce ratio about 2.08 (1.45-2.75). The main extrinsic muscles of the penial complex are a retractor, with a branch attached to the bulb, and another to the caudal end of the penial sheath; and a protractor, with a branch attached to the shoulder of the prepuce and adjoining area of the penial sheath, and another to the caudal end of the penial sheath. Egg capsule C-shaped, with 10-30 elliptical eggs (snails 10mm long) measuring about 1.10 mm (0.90-1.32) through the long axis and surrounded by an inner and an outer lamellate membranes. Jaw a simple obtusely V-shaped plate. radula will be described separately.

Physa cubensis Pfeiffer, 1839 (Pulmonata: Physidae)

A description of Physa cubensis Pfeiffer, 1839, based on 15 speciments collected in Havana, Cuba, is presented. The shell, measuring 9.0 x 4,8mm to 12.3 x 6.4mm, is ovate-oblong, thin, diaphanous, horncolored, shining. Spire elevated, broadly conical; protoconch distinct, roundish, reddish-brown. About five moderately shouldered, roundly convex whorls, penultimate whorl expanded; spiral striation subobsolete; growth line faint on the intermediate whorls, clearly visible on the body whorl, crowded here and there. Suture well impressed. Aperture elongated 2.05 - 2.67 (mean 2.27) times as long as the remaining length of the shell, narrow obovulate-lunate; upper half acute-angled, lower half oval, narrowly rounded at the base; outer lip sharp, inner lip completely closing the umbilical region; a thick callus on the parietal wall; columellar plait well marked. Ratios: shell width/shell length - 0.52-0.61 (mean 0.55); spire length/shell length = 0.27 - 0.33 (mean 0.31); aperture length/shell length = 0.67 - 0.73 (mean 0.69). Oral lappets laterally mucronate; foot spatulate with acuminate tail. Mantle relection with 6 - 8 short triangular dentations in the right lobe (columellar side) and 4 - 6 in the left lobe (near the pneumostome). Renal tube tightly folded into a zigzag course. Ovotestis, ovispermiduct, seminal vesicle, oviduct, nidamental gland, uterus and vagina as in Physa marmorata (see Paraense, 1986, Mem. Inst. Oswaldo Cruz, 81: 459-469). Spermathecal body egg-shaped or pear-shaped; spermathecal ducta uniformly narrow with expanded base, a little longer than the body. Spermiduct, prostate and vas deferens as in P. marmorata (Paraense, loc. cit.). Penis wide proximally, narrowing gradually apicad; penial canal with subterminal outlet. Penial sheath following the width of the penis and ending up by a bulbous expansion somewhat narrower than the proximal portion. Penaial sheath/prepuce ration = 1,25 - 1,83 (mean 1.49). Prepuce much wider than the bulb of the penial shealth, moderately shouldered owing to the intromission of the bulb, and with a large gland in one side of its proximal half occupating about a third of its length. Extrinsic muscles of the penial complex as in P. marmorata. Jaw a simple obtusely V-shaped plate. Radula to be described separetely.

Identification of Mycobacteria by Thin Layer Chromatographic Analysis of Mycolic Acids and Conventional Biochemical Method: Four Years of Experience

Mycolic acids analysis by thin-layer chromatography (TLC) has been employed by several laboratories worldwide as a method for fast identification of mycobacteria. This method was introduced in Brazil by our laboratory in 1992 as a routine identification technique. Up to the present, 861 strains isolated were identified by mycolic acids TLC and by standard biochemical tests; 61% out of these strains came as clinical samples, 4% isolated from frogs and 35% as environmental samples. Mycobacterium tuberculosis strains identified by classical methods were confirmed by their mycolic acids contents (I, III and IV). The method allowed earlier differentiation of M. avium complex - MAC (mycolic acids I, IV and VI) from M. simiae (acids I, II and IV), both with similar biochemical properties. The method also permitted to distinguish M. fortuitum (acids I and V) from M. chelonae (acids I and II) , and to detect mixed mycobacterial infections cases as M. tuberculosis with MAC and M. fortuitum with MAC. Concluding, four years experience shows that mycolic acids TLC is an easy, reliable, fast and inexpensive method, an important tool to put together conventional mycobacteria identification methods.

Plate tectonics, seaways and climate in the historical biogeography of mammals

The marsupial and placental mammals originated at a time when the pattern of geographical barriers (oceans, shallow seas and mountains) was very different from that of today, and climates were warmer. The sequence of changes in these barriers, and their effects on the dispersal of the mammal families and on the faunas of mammals in the different continents, are reviewed. The mammal fauna of South America changed greatly in the Pliocene/Pleistocene, when the newly-complete Panama Isthmus allowed the North American fauna to enter the continent and replace most of the former South American mammal families. Marsupial, but not placental, mammals reached Australia via Antarctica before Australia became isolated, while rats and bats are the only placentals that dispersed naturally from Asia to Australia in the late Cenozoic. Little is known of the early history of the mammal fauna of India. A few mammal families reached Madagascar from Africa in the early Cenozoic over a chain of islands. Africa was isolated for much of the early Cenozoic, though some groups did succeed in entering from Europe. Before the climate cooled in the mid-Cenozoic, the mammal faunas of the Northern Hemisphere were much richer than those of today.

Colorimetric microwell plate reverse-hybridization assay for Mycobacterium tuberculosis detection

Direct smear examination using Ziehl-Neelsen staining for pulmonary tuberculosis (PTB) diagnosis is inexpensive and easy to use, but has the major limitation of low sensitivity. Rapid molecular methods are becoming more widely available in centralized laboratories, but they depend on timely reporting of results and strict quality assurance obtainable only from costly commercial kits available in high burden nations. This study describes a pre-commercial colorimetric method, Detect-TB, for detecting Mycobacterium tuberculosis DNA in which an oligonucleotide probe is fixed onto wells of microwell plates and hybridized with biotinylated polymerase chain reaction amplification products derived from clinical samples. The probe is capable of hybridising with the IS6110 insertion element and was used to specifically recognise the M. tuberculosis complex. When combined with an improved silica-based DNA extraction method, the sensitivity of the test was 50 colony-forming units of the M. tuberculosis reference strain H37Rv. The results that were in agreement with reference detection methods were observed in 95.2% (453/476) of samples included in the analysis. Sensitivity and specificity for 301 induced sputum samples and 175 spontaneous sputum samples were 85% and 98%, and 94% and 100%, respectively. This colorimetric method showed similar specificity to that described for commercially available kits and may provide an important contribution for PTB diagnosis.

Rill erosion on an oxisol influenced by a thin compacted layer

The presence of compacted layers in soils can induce subprocesses (e.g., discontinuity of water flow) and induces soil erosion and rill development. This study assesses how rill erosion in Oxisols is affected by a plow pan. The study shows that changes in hydraulic properties occur when the topsoil is eroded because the compacted layer lies close below the surface. The hydraulic properties that induce sediment transport and rill formation (i.e., hydraulic thresholds at which these processes occur) are not the same. Because of the resistance of the compacted layer, the hydraulic conditions leading to rill incision on the soil surface differed from the conditions inducing rill deepening. The Reynolds number was the best hydraulic predictor for both processes. The formed rills were shallow and could easily be removed by tillage between crops. However, during rill development, large amounts of soil and contaminants could also be transferred.

Use of thin films obtained by plasma polymerization for grain protection and germination enhancement

In this work, preliminary results of the use of hydrophobic thin films obtained by plasma deposition to protect grains and seeds are presented: grains coated by the films did not present biological degradation when stored in a saturated water vapor environment, but had their germination accelerated in the presence of water. A model that explains the difference of behavior of the films when exposed to water in vapor form or in liquid form, based on the formation of microchannels within the film that lead to water uptake in seeds, is presented. The model was successfully tested using quartz crystal measurements, which showed that the microchannels within the films can favor the adsorption and permeation of water when the films are immersed in water.

Um espalhador de baixo custo de fase estacionária em placas para cromatografia em camada delgada

Chromatography is a means of separating mixtures into their several components. TLC, mainly a liquid/solid process, is one of the separation techniques most often used. It is indispensable in laboratories dealing with natural products, organic and analytical chemistry. Commercial chromatography plates are offered at relatively high cost. In this work the construction of a hand-operated plate coater of stationary phases of low cost and good reproducibility is described to be used in teaching laboratories and research.

Plasma polymerized acetaldehyde thin films for retention of volatile organic compounds

The aim of this work is the production and characterization of plasma polymerized acetaldehyde thin films. These films show highly polar species, are hydrophilic, organophilic and easily adsorb organic reactants with CO radicals but only allow permeation of reactants with OH radicals. The good step coverage of films deposited on aluminum trenches is useful for sensor development. Films deposited on hydrophobic substrates may result in a discontinued layer, which allows the use of preconcentration in sample pretreatment. Deposition on microchannels showed the possibility of chromatographic columns and/or retention system production to selectively detect or remove organic compounds from gas flows.