Isolation of Mycobacterium gordonae from poultry slaughterhouse water in São Paulo State, Brazil

Water samples (24 untreated water, 12 treated water and 24 served water) used in different stages of the slaughter process were examined to identify a possible source of pathogenic mycobacteria. The isolates were identified based on microscopy, morphological and biochemical features, mycolic acid analysis and molecular method - PCR-restriction-enzyme analysis. Eighteen mycobacterial strains were isolated from 60 water samples: 11 from untreated water, 5 from treated water and 2 from served water. All mycobacteria isolated were identified as Mycobacterium gordonae and showed the following PRA genotypes: III (27.8%), IV (38.9%) and V (33.3%).

Photodegradation of sparfloxacin and isolation of its degradation products by preparative HPLC

Sparfloxacin, a third generation fluoroquinolone derivative, is a potent antibacterial agent active against a wide range of Gram-positive and Gram-negative organisms including Streptococcus pneuinoniae, Staphylococcus aureus, methicillin resistant S. aureus, Legionella spp., Mycoplasina spp., Chlamydia spp. and Mycobacterium spp. A drawback of fluoroquinolones is their photoreactivity. Sparfloxacin has been studied in terms of therapeutic activities. However, there are few published of analytical methods being applied to sparfloxacin. The aim in this study was to determine the photodegradation products of sparfloxacin, when submitted to UV light, and to characterize two of these products, designated SPAX-PDP1 and SPAX-PDP2. An accelerated study of stability in methanol solution was carried out by exposing a solution of sparfloxacin to UV light (peak wavelength 290 nm) for 36 hours at room temperature. The products were analyzed by NMR spectrophotometry, IR spectrometry and mass spectrophotometry. The results suggest that the products isolated here could be used to estimate the degradation of sparfloxacin in a stability study. However, the low activity exhibited by UV-irradiated sparfloxacin is a source of concern that demands further investigation of the mechanism of its photodegradation mechanism.

Isolation of lignans glycosides from Alibertia sessilis (Vell.) K. Schum. (Rubiaceae) by preparative high-performance liquid chromatography

Enantiomeric aglycone lignans contained in a mixture were separated from a fraction of the extract of the stems of Alibertia sessilis (Vell.) K. Schum. (Rubiaceae) by preparative high-performance liquid chromatography. An efficient and fast separation can be achieved with methanol-water (30:70, v/v). Their structures were identified as (+)-lyoniresinol 3α-O-β-glucopyranoside and (-)-lyoniresinol 3α-O-β-glucopyranoside, being reported for the first time in Rubiaceae.

First isolation of microorganisms from the gut diverticulum of Aedes aegypti (Diptera: Culicidae): New perspectives for an insect-bacteria association

We show for the first time that the ventral diverticulum of the mosquito gut (impermeable sugar storage organ) harbors microorganisms. The gut diverticulum from newly emerged and non-fed Aedes aegypti was dissected under aseptic conditions, homogenized and plated on BHI medium. Microbial isolates were identified by sequencing of 16S rDNA for bacteria and 28S rDNA for yeast. A direct DNA extraction from Ae. aegypti gut diverticulum was also performed. The bacterial isolates were: Bacillus sp., Bacillus subtilis and Serratia sp. The latter was the predominant bacteria found in our isolations. The yeast species identified was Pichia caribbica.

Environmental isolation of black yeast-like fungi involved in human infection

The present study focuses on potential agents of chromoblastomycosis and other endemic diseases in the state of Paraná, Southern Brazil. Using a highly selective protocol for chaetothyrialean black yeasts and relatives, environmental samples from the living area of symptomatic patients were analysed. Additional strains were isolated from creosote-treated wood and hydrocarbon-polluted environments, as such polluted sites have been supposed to enhance black yeast prevalence. Isolates showed morphologies compatible with the traditional etiological agents of chromoblastomycosis, e.g. Fonsecaea pedrosoi and Phialophora verrucosa, and of agents of subcutaneous or systemic infections like Cladophialophora bantiana and Exophiala jeanselmei. Some agents of mild disease were indeed encountered. However, molecular analysis proved that most environmental strains differed from known etiologic agents of pronounced disease syndromes: they belonged to the same order, but mostly were undescribed species. Agents of chromoblastomycosis and systemic disease thus far are prevalent on the human host. The hydrocarbon-polluted environments yielded yet another spectrum of chaetothyrialean fungi. These observations are of great relevance because they allow us to distinguish between categories of opportunists, indicating possible differences in pathogenicity and virulence.

Selective factors involved in oil flotation isolation of black yeasts from the environment

The oil flotation isolation technique has been successfully applied to recover chaetothyrialean black yeasts and relatives from the environment. The selective mechanisms playing a role in isolation are unknown. The fungi concerned are supposed to occupy specialized microniches in nature, taking advantage of (1) oligotrophism. Mineral oil as a main selective agent may be based on (2) hydrophobicity or on (3) assimilation. All three hypotheses are tested in this paper. Results show that cell wall hydrophobicity is unlikely to be a selective factor. Incubation under poor nutrient conditions provides competitive advantage for black yeasts, especially for Exophiala strains, which are subsequently enriched by mineral oil which enhances growth in this group of fungi. Incubation under mineral media and mineral oil can be used as selective factor.

Isolation of toxoplasma gondii from goats from Brazil

Goats are economically important in many countries, and little is known of caprine toxoplasmosis in Brazil. Antibodies to Toxoplasma gondii were assayed in the sera of 143 goats from 3 Brazilian states, using modified agglutination test (MAT titer ≥1:25); 46 (32.2%) tested positive. Samples of brain, heart, diaphragm, and masseter of seropositive animals were pooled, digested in pepsin, and bioassayed in mice. Viable T. gondii specimens were isolated from tissue homogenates of 12 goats; the isolates were designated TgGtBr1-12. Ten of the 12 isolates killed 100 of infected mice, indicating that goats can harbor mouse-virulent T. gondii and, hence, can serve as a source of infection for humans. © 2009 American Society of Parasitologists.

Seduced and abandoned: the social isolation of the urban poor

Includes bibliography

Enzymatic transesterification of soybean oil with ethanol using lipases immobilized on highly crystalline PVA microspheres

Polyvinyl alcohol (PVA) microspheres with different degree of crystallinity were used as solid supports for Rhizomucor miehei lipase immobilization, and the enzyme-PVA complexes were used as biocatalysts for the transesterification of soybean oil to fatty acid ethyl esters (FAEE). The amounts of immobilized enzyme on the polymeric supports were similar for both the amorphous microspheres (PVA4) and the high crystalline microspheres (PVA25). However, the enzymatic activity of the immobilized enzymes was depended on the crystallinity degree of the PVA microspheres: enzymes immobilized on the PVA4 microspheres have shown low enzymatic activity (6.13 U mg-1), in comparison with enzymes immobilized on the high crystalline PVA25 microspheres (149.15 U mg-1). A synergistic effect was observed for the enzyme-PVA25 complex during the transesterification reaction of soybean oil to FAEE: transesterification reactions with free enzyme with the equivalent amount of enzyme that were immobilized onto the PVA25 microspheres (5.4 U) have yielded only 20% of FAEE, reactions with the pure highly crystalline microsphere PVA25 have not yielded FAEE, however reactions with the enzyme-PVA25 complexes have yielded 66.3% of FAEE. This synergistic effect of an immobilized enzyme on a polymeric support has not been observed before for transesterification reaction of triacylglycerides into FAEE. Based on ATR-FTIR, 23Na- and 13C-NMR-MAS spectroscopic data and the interaction of the polymeric network intermolecular hydrogen bonds with the lipases residual amino acids a possible explanation for this synergistic effect is provided. © 2013 Elsevier Ltd. All rights reserved.

The enzymatic recalcitrance of internodes of sugar cane hybrids with contrasting lignin contents

The recalcitrance in grasses varies according to cell type and maturation. The origin of the recalcitrance in different regions from sugar cane internodes with varied lignin contents was evaluated. The efficiency of enzymatic hydrolysis was correlated with the chemical, micromorphological and microspectrophotometric characteristics of the samples. The internodes of three sugar cane hybrids were dissected into four different fractions. The outermost fraction and the rind were the most recalcitrant regions, whereas the pith-rind interface and the pith were less recalcitrant. Cellulose conversion reached 86% after 72h of enzymatic digestion of the pith from the hybrid with the lowest lignin content. There was an inversely proportional correlation between the area occupied by vascular bundles and the efficiency of cellulose hydrolysis. High cellulose and low lignin or hemicellulose contents enhanced the efficiency of enzymatic hydrolysis of the polysaccharides. The critical evaluation of the results permitted to propose an empirical parameter for predicting cellulose conversion levels that accounts for the positive effect of high cellulose and low lignin plus hemicellulose and the detrimental effect of abundant vascular bundles. The cellulose conversion levels fit well to this calculated parameter, following a second order polynomial with an r2 value of 0.96. © 2013 Elsevier B.V.

First isolation of Cryptococcus gattii molecular type VGII and Cryptococcus neoformans molecular type VNI from environmental sources in the city of Belém, Pará, Brazil

Cryptococcus neoformans and Cryptococcus gattii are important agents of meningoencephalitis in humans in the city of Belém. This clinical data suggests that the region may be a highly endemic area for the pathogenic Cryptococcus species within the state of Pará (PA), Northern Brazil. Preliminary analysis of 11 environmental samples from the city of Belém showed two positive locations, including a hollow of a kassod tree (Senna siamea) colonized simultaneously by C. gattii molecular type VGII and C. neoformans molecular type VNI, and a birdcage in a commercial aviary positive for C. neoformans, molecular type VNI. This is the first evidence of an environmental occurrence of molecular types VNI and VGII in PA.

Isolation of Fonsecaea pedrosoi from thorns of Mimosa pudica, a probable natural source of chromoblastomycosis

Este trabalho demonstra o isolamento de Fonsecaea pedrosoi de espinhos da planta Mimosa pudica L., a partir do local de suposta infecção identificado pela paciente infectada. O diagnóstico clínico de cromoblastomicose foi estabelecido pelo achado de corpos fumagóides no exame microscópico direto e pelas culturas de F. pedrosoi do material obtido da lesão da paciente. A mesma espécie foi isolada da paciente e da planta. A microscopia eletrônica de transmissão da superfície dos espinhos evidenciou a disposição conidial característica de F. pedrosoi. Estes dados indicam que a planta M. pudica deve ser uma fonte natural de infecção do fungo F. pedrosoi.

Mechanical isolation of capuchin monkey (Cebus apella) preantral ovarian follicles

O presente trabalho objetivou adaptar um procedimento mecânico para o isolamento de folículos pré-antrais a partir de ovários de Cebus apella. Para isso, foi testado o efeito do intervalo de cortes seriados do tissue chopper sobre o número de folículos pré-antrais isolados a partir de ovários (n=6) de três fêmeas de C. apella, duas pré-púberes e uma adulta. Os ovários foram divididos em quatro partes iguais e fragmentados com auxílio de um tissue chopper, ajustado para a realização de secções seriadas a intervalos de 250, 500, 750 e 1000µm, respectivamente. Os folículos isolados foram contados em câmara de Neubauer e classificados em primordiais, primários e secundários. O número (média ± EP) de folículos pré-antrais isolados de 1/4 de ovário variou de 68.330+17.590, no intervalo de corte de 1.000µm, a 300.830+111.460, no intervalo de corte de 500µm, o de melhores resultados. O diâmetro médio dos folículos pré-antrais isolados variou de 11,6µm a 27, 8µm.

Detection of A/B toxin and isolation of Clostridium difficile and Clostridium perfringens from foals

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)