Estudo da bile por drenagem nasobilar na coledocolitiase apos esfincterotomia endoscopica : descricao de tecnica e analise bacteriologica

Esse trabalho teve como objetivo principal salientar o potencial da drenagem nasobiliar (DNB) como uma forma não cirúrgica de acesso à bile, utilizando como modelo uma téc- nica de DNB no estudo bacteriológico da bile em pesquisa. Para tal, foram estudados 17 pacientes portadores de coledo- colitíase submetidos eletivamente à colangiopancreatografia endoscópica retrógrada na Unidade de Endoscopia do Hospital de Clínicas de Porto Alegre. Foram realizadas DNB por até três dias com coletas seriadas de bile no momento do exame e a cada 24 horas, visando analisar os germes mais prevalentes e o perfil evolutivo da microbiota bacteriana. Correlacionou- se infecção biliar(IB), definida como 105 unidades formadoras de colônias (UFC)/ml de bile, com dados clínico-laboratori- ais obtidos dos prontuários (idade, sexo, presença ou não de febre, icterícia, leucocitose, elevação de fosfatase alca- lina, divertículos justapapilares, uso de antibióticos e co- lecistectomia prévia). A única intercorrência foi desconfor- to na retrofaringe em 28% dos casos. Foram tomadas medidas preventivas visando reduzir a contaminação do sistema. As enterobacteriácias (Klebsiella e E. coli) foram os germes mais encontrados. Ocorreu crescimento bacteriano em 71% dos casos na primeira coleta, embora 30% tivessem IB. Houve al- teração da microbiota biliar em 58% dos casos da primeira para a segunda coleta e em 81% dos casos desta para a terceira. Enquanto IB foi identificada em 30% dos casos na primeira coleta, esta atingiu 50% na segunda, 90% na terceira e 100% na última coleta, embora todos os pacientes tivessem evoluído satisfatóriamente. O perfil bacteriano qualitativo também se alterou, havendo predominância de Klebsiella e E. coli na primeira coleta, acréscimo de Streptococcus faecalis na segunda e apenas Pseudomonas na última. A associação entre IB e os dados clínico-laboratoriais não foi estatisticamente significativa. Concluiu-se que as enterobacteriácias Gram - foram os germes mais prevalentes nos pacientes com coledocolitíase, sendo que o perfil bacteriológico foi significativamente alterado com a DNB, embora sem implicação no quadro clínico. Além disto, não houve associação entre os dados clínico-laboratoriais estudados e a presença de IB.

Aspectos microbiológicos da bile de pacientes com suspeita de coledocolilitíase e suas repercussões no tratamento das infecções biliares

A infecção das vias biliares é uma doença freqüente com alta morbidade e mortalidade, que pode variar de 10 a 60% dependendo de sua gravidade. A causa mais comum desta infecção é a presença de cálculos na via biliar principal que propicia o surgimento de bacteriobilia. O profundo conhecimento das características microbiológicas da bile nos casos de coledocolitíase e infecção das vias biliares são fundamentais para o melhor diagnóstico desta infecção e escolha da antibioticoterapia a ser instituída. Assim, o objetivo deste estudo foi de caracterizar os principais aspectos microbiológicos da bile dos pacientes com e sem coledocolitíase e avaliar sua importância na escolha dos antimicrobianos para o tratamento da infecção das vias biliares. Foram analisados 33 pacientes que foram divididos em um grupo de 10 pacientes sem coledocolitíase (grupo controle) no momento da Colangiografia Endoscópica (CPER) e em outro grupo de 23 pacientes com coledocolitíase. A bile de todos os pacientes foi coletada no início do procedimento endoscópico, através de catater introduzido na via biliar. O exame de microscopia direta com coloração de Gram e as culturas da bile foram negativas nos 10 pacientes que não apresentaram coledocolitíase durante a CPER. Dos 23 pacientes com cálculos na via biliar principal, 19 (83%) apresentaram culturas positivas. Desses 19 pacientes com culturas de bile positivas, 18 (94,7%) apresentaram microorganismos detectáveis à microscopia direta com coloração de Gram. Apenas um paciente apresentou crescimento de germe anaeróbio (Bacteroides fragilis). O cultivo de 28 bactérias teve predominância de microorganismos Gram negativos (18 bactérias- 64,3%). Os germes isolados foram E. coli (9, 32,1%), Klebsiella pneumoniae (5, 17,9%), Enterococcus faecalis (5, 17,9%), Streptococcus alfa-haemoliticus (3, 10,7%), Streptococcus viridans (2, 7,1%), Enterobacter cloacae (2, 7,1%), Panteona aglomerans (1, 3,6%) e Pseudomonas aeruginosa (1, 3,6%). Todos os pacientes com microorganismos detectados pela microscopia direta com coloração de Gram tiveram crescimento bacteriano em suas culturas, por outro lado nenhum paciente com cultura negativa apresentou microoorganismos à microsopia direta ( p= 0,0005). Nesses casos, a microsopia direta apresentou uma especificidade de 100% e sensibilidade de 80%. A análise quantitativa das culturas da bile mostrou que das 19 culturas positivas, 12 (63,2%) tiveram pelo menos um germe com contagem superior a 105 ufc/ml. Todas as bactérias Gram positivas isoladas foram sensíveis à ampicilina, da mesma forma que todas as Gram negativas foram sensíveis aos aminoglicosídeos. Os achados deste estudo demonstram uma boa correlação entre a microscopia direta da bile com coloração de Gram e os achados bacteriológicos das culturas da bile coletada por colangiografia endoscópica retrógrada. O esquema terapêutico antimicrobiano tradicionalmente empregado em nosso hospital, que inclui a combinação de ampicilina e gentamicina, parece ser adequado, pois apresenta eficácia terapêutica contra os principais microorganismos responsáveis pela infecção das vias biliares.

HPLC-DAD methodology for the quantification of organic acids, furans and polyphenols by direct injection of wine samples

This article proposes a simple and sensitive HPLC method with photo-diode array detection for the analysis of organic acids, monomeric polyphenols and furanic compounds in wine samples by direct injection. The chromatographic separation of 8 organic acids, 2 furans and 22 phenolic compounds was carried out with a buffered solution (pH 2.70) and acetonitrile as mobile phases and a difunctionally bonded C18 stationary phase, Atlantis dC18 (250 4.6 mm, 5mm) column. The elution was performed in 12 min for the organic acids and in 60 min for the phenolic compounds, including phenolic acids, stilbenes and flavonoids. Target compounds were detected at 210 nm (organic acids, flavan-3-ols and benzoic acids), 254 nm (ellagic acid), 280 nm (furans and cinnamic acid), 315 nm (hydroxycinnamic acids and trans-resveratrol) and 360 nm (flavonoids). The RSD for the repeatability test (n55) of peak area and retention times were below 3.1 and 0.3%, respectively, for phenolics and below 1.0 and 0.2% for organic acids. The RSDs expressing the reproducibility of the method were higher than for the repeatability results but all below 9.0%. Method accuracy was evaluated by the recovery results, with averaged values between 80 and 104% for polyphenols and 97–105% for organic acids. The calibration curves, obtained by triplicate injection of standard solutions, showed good linearity with regression coefficients higher than 0.9982 for polyphenols and 0.9997 for organic acids. The LOD was in the range of 0.07–0.49 mg/L for polyphenols (cinnamic and gallic acids, respectively) and 0.001–0.046 g/L for organic acids (oxalic and lactic acids, respectively). The method was successfully used to measure and assess the polyphenolic fingerprint and organic acids profile of red, white, rose ´ and fortified wines.

An improved and fast UHPLC-PDA methodology for determination of L-ascorbic and dehydroascorbic acids in fruits and vegetables: evaluation of degradation rate during storage

This study provides a versatile validated method to determine the total vitamin C content, as the sum of the contents of L-ascorbic acid (L-AA) and dehydroascorbic acid (DHAA), in several fruits and vegetables and its degradability with storage time. Seven horticultural crops from two different origins were analyzed using an ultrahigh-performance liquid chromatographic–photodiode array (UHPLC-PDA) system, equipped with a new trifunctional high strength silica (100% silica particle) analytical column (100 mm×2.1 mm, 1.7 μm particle size) using 0.1% (v/v) formic acid as mobile phase, in isocratic mode. This new stationary phase, specially designed for polar compounds, overcomes the problems normally encountered in HPLC and is suitable for the analysis of large batches of samples without L-AA degradation. In addition, it proves to be an excellent alternative to conventional C18 columns for the determination of L-AA in fruits and vegetables. The method was fully validated in terms of linearity, detection (LOD) and quantification (LOQ) limits, accuracy, and inter/intraday precision. Validation experiments revealed very good recovery rate of 96.6±4.4% for L-AA and 103.1±4.8 % for total vitamin C, good linearity with r2-values >0.999 within the established concentration range, excellent repeatability (0.5%), and reproducibility (1.6%) values. The LOD of the method was 22 ng/mL whereas the LOQ was 67 ng/mL. It was possible to demonstrate that L-AA and DHAA concentrations in the different horticulture products varied oppositely with time of storage not always affecting the total amount of vitamin C during shelf-life. Locally produced fruits have higher concentrations of vitamin C, compared with imported ones, but vegetables showed the opposite trend. Moreover, this UHPLC-PDA methodology proves to be an improved, simple, and fast approach for determining the total content of vitamin C in various food commodities, with high sensitivity, selectivity, and resolving power within 3 min of run analysis.

Simultaneous analysis of free amino acids and biogenic amines in honey and wine samples using in loop orthophthalaldeyde derivatization procedure

This work presents a RP-HPLC method for the simultaneous quantification of free amino acids and biogenic amines in liquid food matrices and the results of the application to honey and wine samples obtained from different production processes and geographic origins. The developed methodology is based on a pre-column derivatization with o-phthaldialdehyde carried out in the sample injection loop. The compounds were separated in a Nova-Pack RP-C18 column (150 mm × 3.9 mm, 4 μm) at 35 °C. The mobile phase used was a mixture of phase A: 10 mM sodium phosphate buffer (pH 7.3), methanol and tetrahydrofuran (91:8:1); and phase B: methanol and phosphate buffer (80:20), with a flow rate of 1.0 ml/min. Fluorescence detection was used at an excitation wavelength of 335 nm and an emission wavelength of 440 nm. The separation and quantification of 19 amino acids and 6 amines was carried out in a single run as their OPA/MCE derivatives elute within 80 min, ensuring a reproducible quantification. The method showed to be adequate for the purpose, with an average RSD of 2% for the different amino acids; detection limits varying between 0.71 mg/l (Asn) and 8.26 mg/l (Lys) and recovery rates between 63.0% (Cad) and 98.0% (Asp). The amino acids present at the highest concentration in honey and wine samples were phenylalanine and arginine, respectively. Only residual levels of biogenic amines were detected in the analysed samples.

Histopathology findings in common marmosets (Callithrix jacchus Linnaeus, 1758) with chronic weight loss associated with bile tract obstruction by infestation with Platynosomum (Loos, 1907)

Chronic weight loss in marmosets is often associated with wasting marmoset syndrome (WMS), an important disease that occurs in callitrichid colonies around the world. Even though its etiology is very difficult to determine, particular variables, such as weight loss, diarrhea and alopecia, associated or not with infestation in the pancreatic ducts with Trichospirura leptossoma (Nematoda: Thelazioidea), seem to be linked with the syndrome. This study investigated the histopathology of the lungs, duodenum, liver, gallbladder, extrahepatic bile ducts and pancreatic ducts of six common marmosets (Callithrix jacchus) suffering from severe non-diarrheic weight loss. Three individuals died naturally and the other three were euthanized. Microscopic findings showed the presence of adult flukes (Platynosomum) in the liver. These flukes, which provoke common infection in cats, were also observed inside the gallbladder as well as in the intra and extrahepatic bile ducts in common marmosets. Portal fibrosis was observed in two animals, which developed chronic fibrosing hepatopathy (biliary pattern, grade 3). The disease progresses without diarrhea and without pancreatic lesions or infestation. With the rogression, the animals presented with ascending cholangitis, cholestasis and portal fibrosis, sometimes culminating in secondary biliary cirrhosis. Therefore, this nfirmity, associated with chronic weight loss in common marmosets, could be another tiological factor linked with WMS

Quality of polyunsaturated fatty acids in Nile tilapias (Oreochromis niloticus) fed with vitamin E supplementation

Freshwater fish are an important source of protein, but they also contain other highly nutritive components such as fats. Polyunsaturated fatty acids (PUFAs) are essential for normal growth, development and reproduction of vertebrates. The antioxidant role of vitamin E in cell membranes prevents fatty acid and cholesterol oxidation, thereby promoting PUFA and subcellular particle stabilization. The effects of vitamin E supplementation on the quality of Nile tilapia (Oreochromis niloticus) carcass were investigated. The experiments were carried out in an experimental laboratory over 106 d. After sex reversal, 400 early juvenile O. niloticus were tested in a completely randomized experiment with 5 treatments (4 repetitions each), consisting of vitamin E monophosphate supplementation at 0, 50, 100, 150 or 200 mg/kg of a base diet. Treatment diets contained equal amounts of protein and energy. Tilapias supplemented with vitamin E contained arachidonic acid (20:4 omega-6; AA) which participates in inflammatory response. Nile tilapia carcasses that received vitamin E at 100 and 150 mg/kg diet had improved carcass quality by increasing the PUFA:SFA ratio and had the highest levels of polyunsaturated fatty acids from the omega-3 (linolenic acid; 18:3 omega-3) and omega-6 (linoleic acid; 18:2 omega-6) series. (C) 2012 Elsevier Ltd. All rights reserved.

Prevention of Salmonella infection by contact using intestinal flora of adult birds and/or a mixture of organic acids

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Organic acids and/or compound with defined microorganisms to control Salmonella enterica serovar Enteritidis experimental infection in chickens

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Kinetics of chronic inflammation in Nile tilapia fed n-3 and n-6 essential fatty acids

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Impact of atmosphere, organic acids, and calcium on quality of fresh-cut 'Kensington' mango

Fresh-cut slices from ripe 'Kensington' mango (Mangifera indica L.) were prepared aseptically and stored under various treatments at 3 degrees C. Treatments included reduced oxygen (2.5%), enhanced carbon dioxide (5-40%), organic acid application, calcium chloride application, and combinations of the above. Symptoms limiting shelf-life were characterised by tissue darkening, development of a 'glassy' appearance, surface desiccation, and loss of firmness. Reduced oxygen (2.5%) was effective at controlling tissue darkening and the development of a 'glassy' appearance, while calcium application (3%) was partly effective at controlling darkening. Calcium chloride however significantly slowed (but did not stop) loss of tissue firmness. Carbon dioxide (5-40%) and citric acid had little positive effect on shelf-life, with both treatments appearing to promote tissue softening. A combination of low oxygen and calcium allowed 'Kensington' slices to be held for at least 15 days at 3 degrees C. (C) 2006 Elsevier B.V All rights reserved.

Sunflower meal in commercial layer diets formulated on total and digestible amino acids basis

An experiment was conduced to evaluate the inclusion of sunflower meal (SBM) in commercial layer diets formulated on total or digestible amino acids basis. One hundred forty-four 41-week-old Lohmann LSL layers were distributed in a completely randomized experimental design in a 2 x 4 factorial arrangement with three replications of six birds each. Treatments consisted of a combination of four SBM inclusion levels SBM(0%, 4%, 8%, and 12%) and feed formulation according two amino acid recommendations (total or digestible). The experimental period was divided into five periods of fourteen days. Performance parameters (egg production, feed intake, feed conversion, egg mass) were evaluated for each period. In the last two days of each period, three eggs per replication were collected to evaluate egg quality (Haugh units, specific gravity, egg weight, eggshell thickness, and eggshell percentage). Hens fed on total amino acid recommendation presented the highest values for egg weight. Diets formulated on digestible amino acids basis showed an improvement in eggshell percentage and egg specific gravity. SBM addition in commercial layer diets did not influence performance; however, increasing SBM dietary levels SBM improved eggshell quality.

Commercial laying hen diets formulated according to different recommendations of total and digestible amino acids

An experiment was conducted to evaluate different commercial laying hen diets formulated based on recommendations for total and digestible amino acids. One hundred and twenty Lohmann LSL commercial laying hens aged 25 weeks were distributed in a completely randomized experimental design involving five replications of six birds in four treatments. Diet formulation on a total amino acid basis followed the recommendations of NRC (1994) and Rostagno et al. (2000), whereas formulation on digestible amino acids basis was according to Rostagno et al. (2000) and Degussa (1997) recommendations. The experimental period was divided into five periods of fourteen days. Performance parameters (egg production, feed intake, feed conversion, egg mass) were evaluated for each period, and on the last two days of each period, three eggs per replication were collected to evaluate egg quality parameters (Haugh unit, egg specific gravity, egg weight, eggshell thickness and percentage). Means were compared by orthogonal contrasts. Results on feed intake, egg production, egg mass, feed conversion and egg specific gravity showed that total amino acid recommendations promoted better bird responses than digestible amino acid recommendations.