Isolation and identification of mycobacteria from livestock specimens and milk obtained in Brazil

The prevalence of Mycobacterium bovis and other mycobacterial species in livestock specimens and milk was evaluated. An emphasis was placed upon the distribution of these organisms in milk that is readily available to the public that was either untreated, pasteurized, or treated using ultra high temperature. Twenty-two pathologic specimens from livestock (bovine, swine and bubaline) in five Brazilian states and 128 bovine milk samples from retail markets in the State of São Paulo were examined for mycobacteria. Identification was made by classical biochemical tests, thin layer chromatography of mycolic acids and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. Mycobacteria were isolated from 15 (68.2%) caseous lesions and from 23 (18%) milk samples. Eleven isolates were identified as M. bovis, and the remaining 27 nontuberculous mycobacterial isolates were represented by five species and six unidentified rapidly growing mycobacterial strains. The data demonstrate that animal products in Brazil are frequent reservoirs of mycobacteria and may pose a risk to the public.

Parasitism of two zoonotic reservoirs Dasyprocta leporina and D. fuliginosa (Rodentia) from Amazonas, with Trichostrongylina nematodes (Heligmonellidae): description of a new genus and a new species

A new genus and a new species of Heligmonellidae nematodes are described parasiting the stomach of three agoutis (two Dasyprocta fuliginosa and one D. leporina) captured in the middle and high Negro river microregion, state of Amazonas, Brazil. The new genus, as well as its type-species, are closely related to the trichostrongylids included in Fuellebornema, particularly on what concerns the pattern of the caudal bursa, but differing from them by the characteristics of the synlophe, that presents a poorly developed carene, when compared to the referred number of body ridges in Freitastrongylus n. gen. and consequently in F. angelae n. sp.,in which the ridges are well developed and the carene at mid-body has a similar size when compared to the ridge situated in front of the right field (ridge no. 5). Caudal bursa is of the type 1-4, with rays 9 shorter than rays 10, with a very long genital cone.

Natural Leishmania sp. reservoirs and phlebotomine sandfly food source identification in Ibitipoca State Park, Minas Gerais, Brazil

Leishmania spp are distributed throughout the world and different species are associated with varying degrees of disease severity. However, leishmaniasis is thought to be confined to areas of the world where its insect vectors, sandflies, are present. Phlebotomine sandflies obtain blood meals from a variety of wild and domestic animals and sometimes from humans. These vectors transmit Leishmania spp, the aetiological agent of leishmaniasis. Identification of sandfly blood meals has generally been performed using serological methods, although a few studies have used molecular procedures in artificially fed insects. In this study, cytochrome b gene (cytB) polymerase chain reaction (PCR) was performed in DNA samples isolated from 38 engorged Psychodopygus lloydi and the expected 359 bp fragment was identified from all of the samples. The amplified product was digested using restriction enzymes and analysed for restriction fragment length polymorphisms (RFLPs). We identified food sources for 23 females; 34.8% yielded a primate-specific banding profile and 26.1% and 39.1% showed banding patterns specific to birds or mixed restriction profiles (rodent/marsupial, human/bird, rodent/marsupial/human), respectively. The food sources of 15 flies could not be identified. Two female P. lloydi were determined to be infected by Leishmania using internal transcribed spacer 1 and heat shock protein 70 kDa PCR-RFLP. The two female sandflies, both of which fed on rodents/marsupials, were further characterised as infected with Leishmania (Viannia) braziliensis. These results constitute an important step towards applying methodologies based on cytB amplification as a tool for identifying the food sources of female sandflies.

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.

Persistent organic pollutants in two reservoirs along the Paraíba do Sul-Guandu River system, Rio de Janeiro, Brazil

Sediment contamination is evaluated by determining organic micropollutants (organochlorine compounds - OCs and polycyclic aromatic hydrocarbons - PAHs) in two important Brazilian water reservoirs. Trace levels of OCs were observed in the Santana reservoir (44.8 ng g-1 d.w. of p,p'-DDT), while in the Funil reservoir the levels were below detection level. Forty-eight percent of the found sigmaocs were polychlorinated biphenyls, 29% dichlorodiphenyltrichloroethane (DDT), 18% Drins, and 5% other pesticides (HCB, Heptachlor, Heptachlor-epoxide, gamma-HCH and a-Endosulfan). We observed lower levels of sigmaPAH in the Funil reservoir (1 to 275 ng g-1d.w.) than in the Santana reservoir (2.2 to 26.7 µg g-1 d.w.).

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.

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.

Photometric procedure for quantitative analysis of Aflatoxin B1 in peanuts by thin-layer chromatography using charge coupled device detector

A photometric procedure was developed for determination of aflatoxin B1 in peanuts by TLC-CCD technique. The quantification and detection limit were 1.2 μg kg-1 and 0.4 ng per spot, respectively, with mean recovery of 98%. The CCD camera is sufficiently sensitive to detect small changes in spots fluorescence intensity and the results for performance confirmed the efficiency of the method. Another important property of CCD detector is its linearity for a wide range of luminous stimulus determined by analysis of five-point calibration curves using the intensity of AFB1 fluorescence versus AFB1 concentration (0.8 to 4.8 ng per spot). The method was applied to the analysis of thirty nine peanut samples and aflatoxin B1 levels ranged from 16 to 115 μg kg-1. The TLC-CCD and the photometric procedure developed in this study demonstrated to be a simple and efficient tool for quantitative analyses of AFB1 in peanut samples.

High performance thin layer chromatographic determination of nifedipine in human serum after liquid-liquid extraction

A method using HPTLC for quantitation of nifedipine in serum was developed and validated. It includes a liquid-liquid extraction, and carbamazepine as internal standard. Chloroform: ethyl acetate: cyclohexane (19:2:2, v/v/v) was the mobile phase. The method showed good relationship (r = 0.996) (2.00 to 25.00 ng/band, corresponding to 0.02 and 0.25 ng/µL in serum). The % RSD of intra-assay and inter-assay, were between 0.57 and 3.56 and 1.16 to 3.60, respectively. LOD and LOQ were 0.72 and 0.86 ng/band, respectively. The recovery values were between 93 and 102%. Rf for nifedipine and carbamazepine were 0.31 and 0.10, respectively.

Georeferenced database generation with the purpose of hydrologic molding in reservoirs of the hydrographic basin of Jaguaribe river in the state of Ceará, Brazil

The edafoclimatic conditions of the Brazilian semiarid region favor the water loss by surface runoff. The state of Ceará, almost completely covered by semiarid, has developed public policies for the construction of dams in order to attend the varied water demand. Several hydrological models were developed to support decisive processes in the complex management of reservoirs. This study aimed to establish a methodology for obtaining a georeferenced database suitable for use as input data in hydrological modeling in the semiarid of Ceará. It was used images of Landsat satellite and SRTM Mission, and soil maps of the state of Ceará. The Landsat images allowed the determination of the land cover and the SRTM Mission images, the automatic delineation of hydrographic basins. The soil type was obtained through the soil map. The database was obtained for Jaguaribe River hydrographic basin, in the state of Ceará, and is applicable to hydrological modeling based on the Curve Number method for estimating the surface runoff.

Planktic Cyanobacteria from upper Tietê basin reservoirs, SP, Brazil

Considering the great ecological and sanitary importance of the cyanobacteria and the need of detailed information about these organisms in Brazilian water bodies, the present study aims at contributing towards the knowledge of the cyanobacterial flora of five reservoirs belonging to the upper Tietê Basin, São Paulo: Billings, Guarapiranga, Jundiaí, Pirapora, Ponte Nova and Taiaçupeba. In the past several years, these reservoirs have been submitted to severe environmental deterioration and have repeatedly presented cyanobacterial blooms, including those of toxic species. The samples were collected between 1997 and 2003 either with plankton net (20 µm mesh) or van Dorn's bottle, and preserved with lugol solution or formaldehyde. Some species were isolated and maintained in culture. Forty-eight species of cyanobacteria were identified, with predominance of the order Chroococcales (58%), followed by the orders Oscillatoriales (21%) and Nostocales (21%). Among the 48 studied species, 17 (35%) were considered potentially toxic. The occurrence and biodiversity of the cyanobacteria in each reservoir depend on the environmental conditions. Among the five water bodies, Billings Reservoir presented the most adequate situation for the development of a greater number of species (34), probably due to its high pH values (around 8). Pirapora Reservoir on the other hand, with highest conductivity (445.0 µS cm-1) and lowest Secchi depth values (0.2 m), presented the lowest cyanobacterial biodiversity (14 species).

Floristic surveys of aquatic macrophytes in reservoirs in the Agreste zone of Pernambuco State, Brazil

We surveyed the aquatic macrophytes in reservoirs in Camocim de São Félix in the Agreste zone of Pernambuco State, Brazil. Plants were collected on a bimonthly schedule from November/2008 to May/2009 in four reservoirs, and 33 taxa distributed among 23 families were encountered. The family Cyperaceae had the greatest number of species, followed by Lemnaceae. The predominant biologic form was emergent (33.3%) reflecting the extensive shorelines around the reservoirs analyzed.

Vertical growth phase and positive sentinel node in thin melanoma

Sentinel node (SN) status is the most important prognostic factor for localized melanoma. Usually, patients with Breslow thickness of less than 1.0 mm are not included in SN protocols. However, the literature presents a rate ranging from 3 to 7% of nodal recurrence in thin melanoma. Ulceration, regression and high mitotic rate have been considered to be indications for an SN biopsy. The metastatic potential of the vertical growth phase is uncertain. To correlate pathological features in thin melanoma with SN metastasis, we reviewed 358 patients submitted to SN biopsy. Seventy-seven patients with lesions of 1 mm or smaller were included in the study group. Histological evaluation of the primary tumor included thickness, Clark level, mitotic rate, ulceration, regression, and growth phase. Lymphoscintigraphy was performed on all patients. Lymphatic mapping and gamma probe detection were both used for SN biopsy. Histological examination of SN consisted of hematoxylin-eosin and immunohistochemical staining. Median follow-up was 37 months. Six patients had micrometastases. Statistical analysis by the Fisher test showed that ulceration (P = 0.019), high mitotic rate (P = 0.008) and vertical growth phase (P = 0.002) were positively correlated with micrometastases. If other studies confirm these results, more melanoma patients must be submitted to SN biopsy.

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.

Construction of a supercritical fluid extraction (SFE) equipment: validation using annatto and fennel and extract analysis by thin layer chromatography coupled to image

Abstract The present work describes setting up a laboratory unit for supercritical fluid extraction. In addition to its construction, a survey of cost was done to compare the cost of the homemade unit with that of commercial units. The equipment was validated using an extraction of annatto seeds’ oil, and the extraction and fractionation of fennel oil were used to validate the two separators; for both systems, the solvent was carbon dioxide. The chemical profiles of annatto and fennel extracts were assessed using thin layer chromatography; the images of the chromatographic plates were processed using the free ImageJ software. The cost survey showed that the homemade equipment has a very low cost (~US$ 16,000) compared to commercial equipment. The extraction curves of annatto were similar to those obtained in the literature (yield of 3.8% oil). The separators were validated, producing both a 2.5% fraction of fennel seed extract rich in essential oils and another extract fraction composed mainly of oleoresins. The ImageJ software proved to be a low-cost tool for obtaining an initial evaluation of the chemical profile of the extracts.