Analysis of polycyclic aromatic hydrocarbons in fish: optimisation and validation of microwave-assisted extraction

An accurate and sensitive method for determination of 18 polycyclic aromatic hydrocarbons (PAHs) (16 PAHs considered by USEPA as priority pollutants, dibenzo[a,l]pyrene and benzo[j]fluoranthene) in fish samples was validated. Analysis was performed by microwave-assisted extraction and liquid chromatography with photodiode array and fluorescence detection. Response surface methodology was used to find the optimal extraction parameters. Validation of the overall methodology was performed by spiking assays at four levels and using SRM 2977. Quantification limits ranging from 0.15–27.16 ng/g wet weight were obtained. The established method was applied in edible tissues of three commonly consumed and commercially valuable fish species (sardine, chub mackerel and horse mackerel) originated from Atlantic Ocean. Variable levels of naphthalene (1.03–2.95 ng/g wet weight), fluorene (0.34–1.09 ng/g wet weight) and phenanthrene (0.34–3.54 ng/g wet weight) were detected in the analysed samples. None of the samples contained detectable amounts of benzo[a]pyrene, the marker used for evaluating the occurrence and carcinogenic effects of PAHs in food.

Analysis of polycyclic aromatic hydrocarbons in fish: evaluation of a quick, easy, cheap, effective, rugged, and safe extraction method

QuEChERS method was evaluated for extraction of 16 PAHs from fish samples. For a selective measurement of the compounds, extracts were analysed by LC with fluorescence detection. The overall analytical procedure was validated by systematic recovery experiments at three levels and by using the standard reference material SRM 2977 (mussel tissue). The targeted contaminants, except naphthalene and acenaphthene, were successfully extracted from SRM 2977 with recoveries ranging from 63.5–110.0% with variation coefficients not exceeding 8%. The optimum QuEChERS conditions were the following: 5 g of homogenised fish sample, 10 mL of ACN, agitation performed by vortex during 3 min. Quantification limits ranging from 0.12– 1.90 ng/g wet weight (0.30–4.70 µg/L) were obtained. The optimized methodology was applied to assess the safety concerning PAHs contents of horse mackerel (Trachurus trachurus), chub mackerel (Scomber japonicus), sardine (Sardina pilchardus) and farmed seabass (Dicentrarchus labrax). Although benzo(a)pyrene, the marker used for evaluating the carcinogenic risk of PAHs in food, was not detected in the analysed samples (89 individuals corresponding to 27 homogenized samples), the overall mean concentration ranged from 2.52 l 1.20 ng/g in horse mackerel to 14.6 ± 2.8 ng/ g in farmed seabass. Significant differences were found between the mean PAHs concentrations of the four groups.

New approach for vitamin E extraction in rainbow trout flesh: Application in fish fed commercial and red seaweed-supplemented diets

A vitamin E extraction method for rainbow trout flesh was optimized, validated, and applied in fish fed commercial and Gracilaria vermiculophylla-supplemented diets. Five extraction methods were compared. Vitamers were analyzed by HPLC/DAD/fluorescence. A solid-liquid extraction with n-hexane, which showed the best performance, was optimized and validated. Among the eight vitamers, only α- and γ-tocopherol were detected in muscle samples. The final method showed good linearity (>0.999), intra- (<3.1%) and inter-day precision (<2.6%), and recoveries (>96%). Detection and quantification limits were 39.9 and 121.0 ng/g of muscle, for α-tocopherol, and 111.4 ng/g and 337.6 ng/g, for γ-tocopherol, respectively. Compared to the control group, the dietary inclusion of 5% G. vermiculophylla resulted in a slight reduction of lipids in muscle and, consequently, of α- and γ-tocopherol. Nevertheless, vitamin E profile in lipids was maintained. In general, the results may be explained by the lower vitamin E level in seaweed-containing diet. Practical Applications: Based on the validation results and the low solvent consumption, the developed method can be used to analyze vitamin E in rainbow trout. The results of this work are also a valuable information source for fish feed industries and aquaculture producers, which can focus on improving seaweed inclusion in feeds as a source of vitamin E in fish muscle and, therefore, take full advantage of all bioactive components with an important role in fish health and flesh quality.

Exploring the bioavailability of (poly)phenols from berries and their potential activities in humans

(Poly)phenols are the most widely distributed secondary metabolites, in plants, and, therefore, are regular constituents of human food products. The regular ingestion of (poly)phenol-containing foods has been associated with a reduced risk of acquiring chronic diseases and many studies are currently trying to corroborate this theory. However, the precise contribution of (poly)phenols to disease prevention is still unknown.(...)

A review of mycotoxins in food and feed products in Portugal and estimation of probable daily intakes

Mycotoxins are toxic secondary metabolites produced by filamentous fungi that occur naturally in agricultural commodities worldwide. Aflatoxins, ochratoxin A, patulin, fumonisins, zearalenone, trichothecenes and ergot alkaloids are presently the most important for food and feed safety. These compounds are produced by several species that belong to the Aspergillus, Penicillium, Fusarium and Claviceps genera and can be carcinogenic, mutagenic, teratogenic, cytotoxic, neurotoxic, nephrotoxic, estrogenic and immunosuppressant. Human and animal exposure to mycotoxins is generally assessed by taking into account data on the occurrence of mycotoxins in food and feed as well as data on the consumption patterns of the concerned population. This evaluation is crucial to support measures to reduce consumer exposure to mycotoxins. This work reviews the occurrence and levels of mycotoxins in Portuguese food and feed to provide a global overview of this issue in Portugal. With the information collected, the exposure of the Portuguese population to those mycotoxins is assessed, and the estimated dietary intakes are presented.

Process simulation of lime calcination in mixed feed shaft kilns

Magdeburg, Univ., Fak. für Verfahrens- und Systemtechnik, Diss., 2012

Impact of fish cage-culture on the community structure of zooplankton in a tropical reservoir

We investigated the impact of fish cage culture on the zooplankton community structure in a tropical reservoir. We hypothesized that community abundance is greater near cages and increases over time due to the increase in food availability. Samplings were performed near, upstream and downstream from net cages, and before and after net cage installation. The abundance of zooplankton increased 15 days after the experiment was set up, followed by a reduction and finally increased. Rotifer abundance showed significant differences among sites (p<0.05) and sampling periods (p<0.001). Significant differences were also observed in total zooplankton and cladoceran abundance (p<0.001). The spatial and temporal variation of the physical and chemical variables were indirectly correlated with the structure and dynamic of the zooplankton community, as they indicated the primary production in the environment. Our hypothesis was rejected, since the zooplankton was abundant at the reference site. Only rotifers showed higher abundance near cages, due to the influence of food availability. Community dynamics during the experiment was also correlated to food availability. Our results suggest an impact of fish farming on the zooplankton community.

Diel vertical migration of predators (planktivorous fish larvae) and prey (zooplankton) in a tropical lagoon

ABSTRACT Based on the hypothesis that diel vertical migration (DVM) is a mechanism of predator avoidance, the objective of the present study was to test for the occurrence of DVM in planktivorous fish larvae of Hypophthalmus edentatus (Spix, 1829) (Siluriformes, Pimelodidae) and Plagioscion squamosissimus (Heckel, 1840) (Perciformes, Sciaenidae), and zooplankton (rotifers, cladocerans and copepods) in an isolated tropical lagoon in the floodplain of the Upper Paraná River, Brazil (region of Parque Nacional de Ilha Grande). We investigated spatial overlap between predators (planktivorous fish larvae) and prey (zooplankton), and tested which physical and chemical variables of the water are related to the DVM of the studied communities. We performed nocturnal (8:00 pm and 4:00 am) and diurnal sampling (8:00 am and 4:00 pm) in the limnetic region of the lagoon for six consecutive months, from October 2010 to March 2011, which comprises the reproductive period of the fish species analyzed. During the day the larvae tried to remain aggregated in the bottom of the lagoon, whereas at night they tried to disperse in the water column. Especially for cladocerans, the diel vertical migration is an important behavior to avoid predation larvae of H. edentatus and P. squamosissimus once decreased spatial overlap between secured and its potential predators, which corroborates the hypothesis that DVM is a mechanism of predator avoidance. Although significant correlations were observed between the abiotic factors and WMD of microcrustaceans at certain times of day, the effect of predation of fish larvae on zooplankton showed more important in this environment, because the small depth and isolation not allow great variation of abiotic factors seasonally and between strata the lagoon.

Bioaccumulation of 210Pb and 210Po in fish tissues in a radioactive naturally enhanced area: the Peníscola marsh (Castelló, Spain)

210Pb and 210Po concentration in fish tissues from Peníscola marsh were analyzed, being this area a radioactive naturally enhanced marsh located in the East coast of Spain. Results showed that 210Po accumulation in tissues could reach values ranging from 28±8 Bq kg-1 in muscle of Cyprinus carpio, to 8558±6378 Bq kg-1 in gut content of Chelon labrosus. On the other hand, 210Pb concentrations ranged from 8±4 Bq kg-1 in muscle of Cyprinus carpio, to 475±481 Bq kg-1 in gut content of Chelon labrosus. Bioaccumulation pattern is generally 210Po&210Pb, except in spine, where more 210Pb than 210Po is accumulated. When comparing our samples to those collected as blanks, individuals from Peníscola marsh showed an enrichment in 210Po and 210Pb in their tissues compared to the blanks. Bioaccumulation factors showed that feeding is the major input route of 210Pb and 210Po into the fish body. Highest values of 210Pb and 210Po concentration in tissues were found on Chelon labrosus and Carassius auratus, being Cyprinus carpio the species with the lowest average values of 210Pb and 210Po accumulation.

The benefits of using selenium in the treatment of Chagas disease: prevention of right ventricle chamber dilatation and reversion of Trypanosoma cruzi-induced acute and chronic cardiomyopathy in mice

Cardiac damage is a frequent manifestation of Chagas disease, which is caused by the parasite Trypanosoma cruzi. Selenium (Se) is an essential micronutrient, the deficiency of which has been implicated in the development of cardiomyopathy. Our group has previously demonstrated that Se supplementation prevents myocardial damage during acute T. cruzi infection in mice. In this study, we analyzed the effect of Se treatment in cases of T. cruzi infection using prevention and reversion schemes. In the Se prevention scheme, mice were given Se supplements (2 ppm) starting two weeks prior to inoculation with T. cruzi(Brazil strain) and continuing until 120 days post-infection (dpi). In the Se reversion scheme, mice were treated with Se (4 ppm) for 100 days, starting at 160 dpi. Dilatation of the right ventricle was observed in the infected control group at both phases of T. cruzi infection, but it was not observed in the infected group that received Se treatment. Surviving infected mice that were submitted to the Se reversion scheme presented normal P wave values and reduced inflammation of the pericardium. These data indicate that Se treatment prevents right ventricular chamber increase and thus can be proposed as an adjuvant therapy for cardiac alterations already established by T. cruziinfection.

Effect of Bacillus sphaericus Neide on Anopheles (Diptera: Culicidae) and associated insect fauna in fish ponds in the Amazon

ABSTRACTWe analyzed the effects of Bacillus sphaericus on Anopheles larvae and on the associated insect fauna in fish farming ponds. Five breeding sites in the peri-urban area of the city of Manaus, AM, Brazil, were studied. Seven samples were collected from each breeding site and B. sphaericus was applied and reapplied after 15 days. The samples were made at 24 h before application, 24 h post-application and 5 and 15 days post-application. We determined abundance, larval reduction and larval density for Anopheles, and abundance, richness, Shannon diversity index and classified according to the functional trophic groups for associated insect fauna. A total of 904 Anopheles larvae were collected and distributed into five species. Density data and larval reduction demonstrated the rapid effect of the biolarvicide 24 h after application. A total of 4874 associated aquatic insects belonging to six orders and 23 families were collected. Regression analysis of diversity and richness indicated that the application of the biolarvicide had no influence on these indices and thus no effect on the associated insect fauna for a period of 30 days. B. sphaericus was found to be highly effective against the larvae of Anopheles, eliminating the larvae in the first days after application, with no effect on the associated insect fauna present in the fish ponds analyzed.

ASSESSMENT OF BIOAVAILABILITY OF HEAVY METALS AFTER VERMICOMPOSTING IN THE PRESENCE OF ELECTRONIC WASTE

ABSTRACT Heavy metals contained in electronic waste, if discarded improperly, can become bioavailable after vermicomposting, posing a risk to the environment. Small-scale vermicomposting experiments were carried out with printed circuit boards (PCBs) to investigate the migration of heavy metals (Cu, Pb, Zn, Ni, and Sn) to the final compost, as well as the mobility and bioavailability of these metals. High total levels of Pb, Sn and Cu in samples of manure with electronic waste (MEW) and vegetables with electronic waste (VEW) were detected. Based on the initial metal levels in the PCBs and their concentration in the resulting compost, the order of migration of these metals to the MEW and VEW samples was Sn (23.1 %)>Pb (18.4 %)>Ni (4.63 %)>Zn (0.46 %)>Cu (0.14 %) and Sn (24.3 %)>Pb (23.6 %)>Ni (11.33 %)>Zn (1.76 %)>Cu (0.60 %), respectively. Mobility and bioavailability of these metals in the compost were evaluated by three-stage sequential extraction, where F1 was the exchangeable fraction, F2 the organic fraction and F3 the residual fraction. The bioavailability factor (BF) was calculated by the ratio of the sum of fractions F1 and F2 divided by the total sum of the fractions (F1 + F2 + F3). The highest bioavailability factor (BF = 0.92) was found for Pb, the heavy metal considered the greatest environmental concern in this study, indicating the high mobility and the possibility of becoming bioavailable of this metal.

Bioavailability of repeated oral administration of MDL 100,240, a dual inhibitor of angiotensin-converting enzyme and neutral endopeptidase in healthy volunteers.

BACKGROUND: MDL 100,240 (pyrido[2,1-a] [2]benzazepine-4-carboxylic acid,7-[[2-(acetylthio)-1-oxo-3-phenylpropyl]amino]-1,2,3,4,6,7,8, 12b-octahydro-6-oxo, [4S-[4alpha,7alpha(R(*)),12bbeta]]-) is a molecule possessing an inhibiting ability on both angiotensin converting enzyme (ACE) and neutral endopeptidase, the enzyme responsible for atrial natriuretic peptide (ANP) degradation. Such a dual mechanism of action presents a potential clinical interest for the treatment of hypertension and congestive heart failure. OBJECTIVES: To evaluate the bioavailability of MDL 100,240 and its accumulation over repeated oral administration, using ACE inhibition as a surrogate for plasma drug level and determining its profile after oral and i.v. administration. METHODS: First, in an open, one-period, single-dose study, the ACE inhibition profile was characterised following a 12.5 mg MDL 100,240 i.v. infusion. Second, in a three-group, parallel, randomised, double-blind study, each group of four subjects received q.d., over 8 days, 2.5, 10 or 20 mg of MDL 100,240 orally. The ACE inhibition profile was determined on day 1 and day 8. Trough plasma ACE was measured on days 2, 3 and 4. The recovery of ACE activity was monitored up to 72 h after the last dose of MDL 100,240. RESULTS: ACE inhibition profile was similar on day 1 and day 8, and trough inhibition remained unchanged after the 8 days of treatment with 10 mg or 20 mg. Following repeated 2.5-mg ingestion, trough inhibition increased from 33% to 44% after the eighth dose. The oral bioavailability of MDL 100,240 was estimated at 85%, not statistically different from 100%. The accumulation ratio at steady state was estimated at 112%. Expressing the accumulation ratio in terms of half-life, a t(1/2) of 0.31 days or 7. 5 h was estimated. CONCLUSION: MDL 100,240 (oral solution) has a good bioavailability, as estimated by ACE inhibition, and no drug accumulation seems to occur over 8 days with the 10-mg and 20-mg doses, but a slight rise in the trough level is observed with the 2. 5-mg dose.

Allocation of added selenium in lettuce and its impact on roots

Selostus: Seleenin jakautuminen salaatin versoihin ja vaikutus juuriin