Nitrogen metabolism in lactating goats fed with diets containing different protein sources.

This study aimed to evaluate urea excretion, nitrogen balance and microbial protein synthesis in lactating goats fed with diets containing different protein sources in the concentrate (soybean meal, cottonseed meal, aerial part of cassava hay and leucaena hay). Four Alpine goats whose mean body weight was 42.6±6.1 kg at the beginning of the experiment, a mean lactation period of 94.0±9.0 days and a production of 1.7±0.4 kg of milk were distributed in a 4×4 Latin square with four periods of 15 days. Diets were formulated to be isonitrogenous, containing 103.0 g/kg of CP, 400 g/kg of Tifton 85 hay and 600 g/kg of concentrate. Diet containing cottonseed meal provided (p<0.05) increased excretion of urea and urea nitrogen in the urine (g/d and mg/kg of BW) when compared with leucaena hay. The diets affected the concentrations of urea nitrogen in plasma (p<0.05) and excretion of urea nitrogen in milk, being that soybean meal and cottonseed meal showed (p<0.05) higher than the average aerial part of the cassava hay. The use of diets with cottonseed meal as protein source in the concentrate in feeding of lactating goats provides greater nitrogen excretion in urine and negative nitrogen balance, while the concentrate with leucaena hay as a source of protein, provides greater ruminal microbial protein synthesis.

GC-MS determination of malondialdehyde, acrolein, and 4-hydroxy-2-nonenal by ultrasound-assisted dispersive liquid-liquid microextraction in beverages.

A new, fast, simple, and effective ultrasound-assisted dispersive liquid-liquid microextraction procedure (UA-DLLME) for the gas chromatography-mass spectrometry (GC-MS) determination of malondialdehyde, acrolein, and 4-hydroxy-2-nonenal in beverages was successfully developed. 2,4-Dinitrophenylhydrazine derivatization was performed during extraction. An asymmetrical 3541//18 screening design and a central composite surface response design were used to investigate the influence of the most critical factors during the extraction process (ultrasound time and temperature, extraction and disperser solvents volumes, salt addition, and derivatization reagent concentration). According to FDA guidelines, the method was validated, achieving good linearities with r2 ≥ 0.9982, recoveries between 94.0 and 102.4%, and reproducibility with RSD lower than 4.5%. The method was applied to simultaneously determine the compounds in 60 different beverage samples, including beer, coffee, black tea, and fruit juices. The presence of secondary lipid oxidation products is demonstrated in beverages with a strong roasting process or oxidation.

Determination of malondialdehyde, acrolein and four other products of lipid peroxidation in edible oils by Gas-Diffusion Microextraction combined with Dispersive Liquid-Liquid Microextraction.

A new and sensitive analytical method for the simultaneous determination of secondary lipid peroxidation aldehydes has been successfully developed and validated. Malondialdehyde, acrolein, formaldehyde, acetaldehyde, propanal, and pentanal were extracted and derivatized using 2,4-dinitrophenylhydrazine (DNPH) by gas-diffusion microextraction (GDME) combined with dispersive liquid-liquid microextraction (DLLME) for gas chromatography-mass spectrometry (GC-MS) analysis. The experimental conditions have been optimized by experimental designs. The analytical method validation, in accordance to the Food and Drug Administration (FDA) guidance, provided good results in terms of linearity with r2≥0.9974, in the range from 0.15 or 0.3 µg·g-1 to 3 µg·g-1. Limits of detection and limits of quantification were 0.05 or 0.10 and 0.15 or 0.3 µg·g-1, respectively. Precision was tested as a relative standard deviation (RSD≤ 9.5%) and recoveries were between 95% and 110%. The method was applied in the characterization of aldehydes in forty-eight edible oil samples; with the highest concentration found in pomace olive oil for malondialdehyde at 6.64 µg·g-1.

Time domain add-drop multiplexing scheme enhanced using a saw-tooth pulse shaper.

We experimentally demonstrate the use of saw-tooth optical pulses, which are shaped using a fiber Bragg grating, to achieve robust and high performance time-domain add-drop multiplexing in a scheme based on cross-phase (XPM) modulation in an optical fiber, with subsequent offset filtering. As compared to the use of more conventional pulse shapes, such as Gaussian pulses of a similar pulse width, the purpose-shaped saw-tooth pulses allow higher extinction ratios for the add and drop windows and significant improvements in the receiver sensitivity for the dropped and added channels.

Low pressure ion pair chromatography with amperometric detection for the determination of trigonelline in coffee samples.

In this work, the use of ion pair chromatography strategy in low pressure chromatographic flow systems is explored for the first time. The straightforward flow manifold encompassed a peristaltic pump, an injection valve and a 1 cm-length C18 monolithic column. The amperometric detection system relied on a boron-doped diamond electrode, used as working electrode. The determination of trigonelline in coffee samples was the case-study selected. This alkaloid is an important quality marker for this commodity and is usually determined using HPLC-UV methodologies. The proposed methodology, based on ion-pair chromatography with amperometric detection, enabled the quantitative resolution of the studied analyte from the matrix compounds by adding to the mobile phase the ion pair reagent, 1-tetradecanosulfonate sodium. The present work, following the recent developments of the low pressure chromatography approach, demonstrates the potentialities of coupling monolithic columns to traditional flow analysis systems for separation and quantification of ionic or ionisable compounds.

Development of a partitioned liquid-liquid extraction- dispersive solid phase extraction procedure followed by liquid chromatography-tandem mass spectrometry for analysis of 3-monochloropropane-1,2-diol diesters in edible oils.

A fast and effective method using a modified QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) approach which includes partitioned liquid-liquid extraction (PLLE) and dispersive solid phase extraction (dSPE) clean-up step for the determination of seven 3-monochloropropane-1,2-diol (3-MCPD) fatty acid diesters in vegetable oils is developed and validated according to the Food and Drug Administration (FDA) guidelines. Due to the complexity of the matrices, combination of silica based sorbents (Silica Strong Anion Exchange (Si-SAX), Supel™ QuE Z-Sep+ (Z-Sep+) and Primary Secondary Amine (PSA) were tested for lipid removal. The effect of several experimental factors on the efficiency of the extraction procedure was studied by a screening design 3422//16 and a response surface Doehlert design. The separation and determination was carried out by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). The method provided suitable linearity (r2 > 0.9960), precision (relative standard deviation (RSD) lower than 10%) and accuracy, in terms of recovery. The limits of detection (LOD) and limits of quantification (LOQ) ranged from 10 to 20 µg kg -1 and from 25 to 50 µg kg-1, respectively. The recoveries at three spiking levels of 100, 250, and 500 µg kg-1 were over the range of 71.4-122.9% with RSD lower than 13%. The method was successfully applied in edible oils and fatty food samples. The results provide valuable information to assess the risk of exposure to these foodborne contaminants.

Detecting and modelling delayed density-dependence in abundance time series of a small mammal (Didelphis aurita).

We study the population size time series of a Neotropical small mammal with the intent of detecting and modelling population regulation processes generated by density-dependent factors and their possible delayed effects. The application of analysis tools based on principles of statistical generality are nowadays a common practice for describing these phenomena, but, in general, they are more capable of generating clear diagnosis rather than granting valuable modelling. For this reason, in our approach, we detect the principal temporal structures on the bases of different correlation measures, and from these results we build an ad-hoc minimalist autoregressive model that incorporates the main drivers of the dynamics. Surprisingly our model is capable of reproducing very well the time patterns of the empirical series and, for the first time, clearly outlines the importance of the time of attaining sexual maturity as a central temporal scale for the dynamics of this species. In fact, an important advantage of this analysis scheme is that all the model parameters are directly biologically interpretable and potentially measurable, allowing a consistency check between model outputs and independent measurements.

Evaluation of low doses BPA-induced perturbation of glycemia by toxicogenomics points to a primary role of pancreatic islets and to the mechanism of toxicity.

Epidemiologic and experimental studies have associated changes of blood glucose homeostasis to Bisphenol A (BPA) exposure. We took a toxicogenomic approach to investigate the mechanisms of low-dose (1 × 10(-9 )M) BPA toxicity in ex vivo cultures of primary murine pancreatic islets and hepatocytes. Twenty-nine inhibited genes were identified in islets and none in exposed hepatocytes. Although their expression was slightly altered, their impaired cellular level, as a whole, resulted in specific phenotypic changes. Damage of mitochondrial function and metabolism, as predicted by bioinformatics analyses, was observed: BPA exposure led to a time-dependent decrease in mitochondrial membrane potential, to an increase of ROS cellular levels and, finally, to an induction of apoptosis, attributable to the bigger Bax/Bcl-2 ratio owing to activation of NF-κB pathway. Our data suggest a multifactorial mechanism for BPA toxicity in pancreatic islets with emphasis to mitochondria dysfunction and NF-κB activation. Finally, we assessed in vitro the viability of BPA-treated islets in stressing condition, as exposure to high glucose, evidencing a reduced ability of the exposed islets to respond to further damages. The result was confirmed in vivo evaluating the reduction of glycemia in hyperglycemic mice transplanted with control and BPA-treated pancreatic islets. The reported findings identify the pancreatic islet as the main target of BPA toxicity in impairing the glycemia. They suggest that the BPA exposure can weaken the response of the pancreatic islets to damages. The last observation could represent a broader concept whose consideration should lead to the development of experimental plans better reproducing the multiple exposure conditions.

Endothelium-dependent and endothelium-independent effects of adenosine diphosphate in renovascular hypertension.

Norepinephrine-induced responses in isolated perfused mesenteric vascular bed from normotensive and renovascular hypertensive rats were examined in the presence of adenosine diphosphate (ADP, 2 x 10(-6) M). Responses to norepinephrine were significantly greater in vessels from hypertensive rats. Norepinephrine-induced contractions increased after the removal of endothelium. N omega-Nitro-L-arginine (L-NOARG), a potent inhibitor of nitric oxide formation, similarly increased contractions. The greatest responses were obtained, however, after treatment of the vascular segments with methylene blue. The presence of ADP caused significant endothelium-dependent decreases in contractions. Although decreases caused by ADP in vessels with endothelium after treatment with L-NOARG were not statistically significant, a tendency to decreased responses seems to suggest that L-NOARG diminishes but does not completely prevent the effect of ADP in mesenteric vessels. Methylene blue partially reduced the endothelium-dependent ADP-induced relaxant effects in sham-operated nephrectomized rats. A tendency to increased contractions to norepinephrine was observed in the presence of ADP after removal of endothelium. Thus, in the mesenteric resistance arteries of the rat under stimulation by ADP, it appears that nitric oxide released from L-arginine and the activity of soluble guanylate cyclase account only in part for the endothelium-dependent decreased responses to norepinephrine. When nitric oxide formation or soluble guanylate cyclase activity are depressed simultaneously with endothelium damage, ADP released from platelets or red blood cells may be an important factor that acts synergically with vasoconstrictor stimuli.(ABSTRACT TRUNCATED AT 250 WORDS)

Diagnostic evaluation of PCR in goats experimentally infected with Trypanosoma vivax.

Six goats were experimentally infected with a stock of Trypanosoma vivax. Parasitaemia was weekly monitored by buffy coat and wet blood film examination during a period of 15 weeks and another 3 weeks following drug-treatment. Dried blood samples were tested by the polymerase chain reaction (PCR), using an extraction method with Chelex 100 (BioRad). PCR proved consistently more sensitive than the parasitological techniques.

Determination of E-2-nonenal by high-performance liquid chromatography with UV detection assay for the evaluation of beer ageing.

The analysis of E-2-nonenal is of considerable interest for the brewery industry as this compound is claimed to be responsible for a paper/cardboard unpleasant flavour. Usually, the presence of E-2-nonenal can be noticed in aged beers at levels higher than 0.1 microg/l. In this work, an analytical method was developed to determine E-2-nonenal in beer involving steam distillation of beer followed by an extraction/concentration step using solid-phase extraction and determination of E-2-nonenal by HPLC with UV detection. Fastness and simplicity are the main advantages of the proposed method, when compared with other existing methodologies for the determination of E-2-nonenal in beer. Using the developed conditions, the interference of E-2-nonenal formed by degradation of its precursors during steam distillation is almost negligible. The presence of sulphur dioxide at legal levels does not interfere with the assay. The method was used in a comparative study of fresh and either naturally or forced aged beers. A much larger chromatographic peak was found near the peak of E-2-nonenal that correlates well with the peak of E-2-nonenal. Identification of the corresponding compound is currently under investigation, considering its future application on the evaluation of beer ageing.

Simultaneous determination of E-2-nonenal and beta-damascenone in beer by reversed-phase liquid chromatography with UV detection.

A method for the simultaneous determination of E-2-nonenal and beta-damascenone in beer by reversed-phase liquid chromatography using UV detection is presented. The method consists of beer steam distillation, followed by an extraction/concentration step using Sep-Pak Plus C18 RP cartridges and determination by HPLC at 226 nm UV-absorption maximum. The identity of the compounds was confirmed by GC analysis with MS detection of the isolated fractions. A recovery factor of approximately 80% was obtained for beta-damascenone with a R.S.D. of 3%. E-2-Nonenal and beta-damascenone were monitored in a comparative study of fresh and either naturally and forced aged beer. The results obtained show that both compounds have a similar behaviour through an extended storage of beer and consequently can be used as good analytical markers of beer ageing. Nevertheless, the use of beta-damascenone seems to be more convenient because this compound appears in beer in higher concentrations than E-2-nonenal, thus making it easier to measure.

PCR primer evaluation for the detection of trypanosome DNA in naturally infected goats.

The buffy coat of 76 roaming goats from the Bansang and Missira regions in Gambia, was examined for the presence of trypanosomes. From these animals, extractions from dry blood samples on filter paper were subjected to PCR using three different primer sets, ORPHON5J, GOL and TVW, specific for Trypanosoma brucei/Trypanosoma evansi, Trypanosoma congolense and Trypanosoma vivax, respectively. PCR results for T. congolense were 100% concordant with buffy coat examination. Besides the three T. vivax buffy coat-positive samples, another 15 yielded positive with the TVW primers. The ORPHON5J primers yielded no positive results. Analyses with the GOL primers of putatively negative samples, yielded aberrant band patterns whose diagnostic significance still remains to be determined.

Prevalence and incidence of trypanosomosis in horses and donkeys in the Gambia.

In a study of the prevalence and incidence of trypanosomosis in horses and donkeys in two regions of the Gambia, surveys were carried out at Niamina east and Bansang south with a high and low to moderate tsetse challenge, respectively. Eleven horses and 67 donkeys were sampled monthly from August 1997 to September 1998. Blood samples were examined for trypanosomes using the buffy-coat (BC) method and polymerase chain reaction (PCR). Three primer sets were used, specific for either Trypanosoma vivax (TVW), Trypanosoma congolense (GOL) or Trypanosoma brucei (ORPHON5J). The BC results showed that the prevalence (August 1997) and the average monthly incidence (September 1997-1998) of trypanosome infections in horses (45.5 and 16%, respectively) were significantly higher than in donkeys (6.2 and 9%, respectively). Using PCR, the number of detected cases was seven times higher than using the BC. T. congolense was the most frequently observed species, followed by T. vivax and T. brucei. This study confirms earlier observations by other authors that donkeys, which are exposed to a similar tsetse challenge as horses, are significantly less infected with trypanosomes than the latter.