Combined effects of dietary Laminaria digitata with alginate lyase on plasma metabolites and hepatic lipid, pigment and mineral composition of broilers.

BACKGROUND: The Laminaria digitata is an abundant macroalga and a sustainable feedstock for poultry nutrition. L. digitata is a good source of essential amino acids, carbohydrates and vitamins, including A, D, E, and K, as well as triacylglycerols and minerals, in particular iron and calcium. However, the few studies available in the literature with broilers document the application of this macroalga as a dietary supplement rather than a feed ingredient. No study has addressed up until now the effects of a high-level incorporation (> 2% in the diet) of L. digitata on plasma biochemical markers and hepatic lipid composition, as well as minerals and pigments profile in the liver of broilers. Our experimental design included one hundred and twenty Ross 308 male birds contained in 40 wired-floor cages and distributed to the following diets at 22 days of age (n = 10) for 15 days: 1) a corn-soybean basal diet (Control); 2) the basal diet plus 15% of L. digitata (LA); 3) the basal diet plus 15% of L. digitata with 0.005% of Rovabio® Excel AP (LAR); and 4) the basal diet plus 15% of L. digitata with 0.01% of the recombinant CAZyme, alginate lyase (LAE). RESULTS: L. digitata compromised birds' growth performance by causing a reduction in final body weight. It was found an increase in hepatic n-3 and n-6 fatty acids, in particular C18:2n-6, C18:3n-6, C20:4n-6, C20:5n-3, C22:5n-3 and C22:6n-3 with the addition of the macroalga, with or without feed enzymes, to the broiler diets. Also, the beneficial C18:3n-3 fatty acid was increased by combining L. digitata and commercial Rovabio® Excel AP compared to the control diet. The sum of SFA, MUFA and the n-6/n-3 PUFA ratio were decreased by L. digitata, regardless the addition of exogenous enzymes. ß-carotene was enhanced by L. digitata, individually or combined with CAZymes, being also responsible for a positive increase in total pigments. Macrominerals, in particular phosphorous and sulphur, were increased in the liver of broilers fed L. digitata individually relative to the control. For microminerals, copper, iron and the correspondent sum were consistently elevated in the liver of broilers fed L. digitata, individually or combined with exogenous CAZymes. The powerful discriminant analysis tool based on the hepatic characterization revealed a good separation between the control group and L. digitata diets but failed to discriminate the addition of feed enzymes. CONCLUSIONS: Overall, this study highlights the value of L. digitata as a feed ingredient for the poultry industry. Moreover, we can conclude that the effect of L. digitata overpowers the effect of feed enzymes, both the Rovabio® Excel AP and the alginate lyase. Having in mind the negative effects observed on birds' performance, our main recommendation at this stage is to restraint L. digitata incorporation level in forthcoming nutritional studies.

Muscle mineral profile of water buffaloes (Bubalus bubalis) reared in different production systems of the Brazilian Eastern Amazon.

Healthy food must have an adequate balance of macroelements, such as calcium or phosphorus and, microelements, such as iron, copper. This study aimed to evaluate the influence of three extensive systems, during the dry and rainy seasons, and an intensive (feedlot) system in the Eastern Amazon, on the muscle mineral profile of water buffaloes. In total, 12 male buffaloes, aged between 24 and 36 months, slaughtered in commercial slaughterhouses, were used in each of the systems considered: Marajó island, Santarém, Nova Timboteua, and a feedlot. Approximately 5 g of muscle was collected, stored, and frozen, until freeze-dried. The samples were analyzed for the mineral profile using inductively coupled plasma-optical emission spectrometry (ICP-OES). There were significant differences (P < 0.05) for concentrations of sodium (Na), magnesium (Mg), phosphorus (P), sulfur (S), copper (Cu), zinc (Zn), and iron (Fe). Extensive and intensive systems showed significant differences (P < 0.05) for Na, Ca, S, Cu, and Fe concentrations. The season also influenced (P < 0.05) K, Ca, P, S, Zn, and Fe concentrations. The location and season of the year had a significant interaction (P < 0.05) for K, Mg, P, Zn, and Fe concentrations. The study showed that the different Amazonian production systems and the year season influenced the levels of minerals present in buffalo muscle. The values obtained were, in general, higher in extensive production systems, and Marajó Island stood out with higher mineral values in the dry season. Therefore, the meat from animals reared in these systems is a good mineral source for daily human needs.

Mineral Content of Liver of Buffaloes (Bubalus bubalis) Reared in Different Ecosystems in the Eastern Amazon.

This study aimed to evaluate the influence of different production ecosystems, three in native and cultivated pastures (extensive), at two seasons of the year (dry and rainy), and one in confinement (intensive) in the Eastern Amazon, on the mineral content of buffalo liver raised on these ecosystems. Twelve male buffalo (n = 12), aged between 24 and 36 months, slaughtered in commercial slaughterhouses, were used in each of the ecosystems considered: Marajó; Lower Amazon; Cultivated Pasture, and in confinement system, Pará, Brazil. Approximately 5 g of liver was collected, stored and frozen until lyophilization. Samples were analyzed for mineral content based on inductively coupled plasma optical emission spectrometry (ICPOES) readings. The relationship between extensive ecosystems and an intensive production system (p < 0.05) in the values of potassium (K), iron (Fe), copper (Cu), manganese (Mn) and barium (Ba) was evaluated. The different ecosystems studied influenced (p < 0.05) the mineral values found in the liver of buffaloes raised in the ecosystems, for sodium (Na), K, calcium (Ca), magnesium (Mg), sulfur (S), copper (Cu), iron (Fe), manganese (Mn) and barium (Ba). The period of the year interacted with the values of Na, K, S and Cu; however, an interaction of local vs. period of the yer was observed for the values of K, Mg, P, S and Cu. It can be concluded that the buffalo liver is an excellent source of minerals and can be included in the human diet and that the ecosystem the animals are raised influences its content.

Antioxidative response of lettuce (Lactuca sativa) to carbamazepine-induced stress.

Carbamazepine (CBZ) is a widely used anti-epileptic drug that has been detected in wastewaters from sewage treating plants and thus appears in rivers, streams and other water bodies. As plants can absorb this compound, it can also appear in edible plants like lettuce, entering the food chain. In this study, the effect of carbamazepine in lettuce plants grown in hydroponic solution is analyzed. CBZ was detected both in roots and in leaves and is shown to induce oxidative stress. Hydrogen peroxide levels increased both in leaves and in roots while malondialdehyde increased only in leaves. Regarding the activity of antioxidative enzymes in the leaves, it is shown that superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (GPOD) and ascorbate peroxidase (APX) have a relevant role in quenching reactive oxygen species induced by oxidative stress. In roots, the only enzymes that showed increased activity were CAT, GPOD and glutathione reductase (GR). Ascorbate and glutathione also appear to have an important role as antioxidants in response to increased concentrations of carbamazepine. Although the roots are in direct contact with the contaminant, the leaves showed the strongest oxidative effects.

Efficient regulation of copper homeostasis underlies accession-specific sensitivities to excess copper and cadmium in roots of Arabidopsis thaliana.

The commonly used Arabidopsis thaliana natural accessions Columbia (Col-0) and Wassilewskija (Ws) are known to differ in their metal sensitivity, with Col-0 being more sensitive to copper (Cu) and cadmium (Cd) than Ws. As both Cu and Cd are known to affect Cu homeostasis, it was investigated whether this process is part of an accession-specific mechanism underlying their difference in metal sensitivity. As roots are the first contact point during metal exposure, responses were compared between roots of both accessions of hydroponically grown plants exposed to excess Cu or Cd for 24 and 72 h. Root Cu levels increased in both accessions under Cu and Cd exposure. However, under Cu exposure, the downregulation of Cu transporter (COPT) genes in combination with a more pronounced upregulation of metallothionein gene MT2b indicated that Ws plants coped better with the elevated Cu concentrations. The Cd-induced disturbance in Cu homeostasis was more efficiently counteracted in roots of Ws plants than in Col-0 plants. This was indicated by a higher upregulation of the SPL7-mediated pathway, crucial in the regulation of the Cu homeostasis response. In conclusion, maintaining the Cu homeostasis response in roots is key to accession-specific differences in Cu and Cd sensitivity.

Accession-specific life strategies affect responses in leaves of Arabidopsis thaliana plants exposed to excess Cu and Cd.

The natural accession Columbia (Col-0) is considered as the reference genome of the model plant Arabidopsis thaliana. Nonetheless, Col-0 plants are more sensitive to excess copper (Cu) and cadmium (Cd) than other widely used accessions such as Wassilewskija (Ws) plants. In the current study, this accession-specific metal sensitivity is further explored by comparing the responses in leaves of Col-0 and Ws plants exposed to excess Cu and Cd. Our results suggest that different life strategies favored by both accessions under physiological conditions affect their response to metal exposure. While Col-0 plants mainly invest in metal detoxification, Ws plants center on nutrient homeostasis. In particular, the higher expression of genes related to Cu homeostasis genes in non-exposed conditions indicates that Ws plants possess a constitutively efficient metal homeostasis. On the other hand, oxidative stress-related MAPK signaling appears to be boosted in leaves of Col-0 plants exposed to excess Cu. Furthermore, the upregulation of the glutathione (GSH) biosynthesis GSH2 gene and the increased GSH concentration after Cd exposure suggest the activation of detoxification mechanisms, such as phytochelatin production, to counteract the more severe Cd-induced oxidative stress in leaves of Col-0 plants. Exposure to Cd also led to a more pronounced ethylene signaling response in leaves of Col-0 as compared to Ws plants, which could be related to Cd-induced GSH metabolism. In conclusion, accession-specific life strategies clearly affect the way in which leaves of A. thaliana plants cope with excess Cu and Cd.

Characterization of aspartate aminotransferase isoenzymes from leaves of Lupinus albus L. cv Estoril.

Two aspartate aminotransferase (EC 2.6.1.1) isoenzymes (AAT-1 and AAT-2) from Lupinus albus L. cv Estoril were separated, purified, and characterized. The molecular weight, pI value, optimum pH, optimum temperature, and thermodynamic parameters for thermal inactivation of both isoenzymes were obtained. Studies of the kinetic mechanism, and the kinetics of product inhibition and high substrate concentration inhibition, were performed. The effect of some divalent ions and irreversible inhibitors on both AAT isoenzymes was also studied. Native PAGE showed a higher molecular weight for AAT-2 compared with AAT-1. AAT-1 appears to be more anionic than AAT- 2, which was suggested by the anion exchange chromatography. SDS-PAGE showed a similar sub-unit molecular weight for both isoenzymes. The optimum pH (between 8.0 and 9.0) and temperature (60-65 degrees C) were similar for both isoenzymes. In the temperature range of 45-65 degrees C, AAT-2 has higher thermostability than AAT-1. Both isoenzymes showed a high affinity for keto-acid substrates, as well as a higher affinity to aspartate than glutamate. Manganese ions induced an increase in both AAT isoenzymes activities, but no cooperative effect was detected. Among the inhibitors tested, hydroxylamine affected both isoenzymes activity by an irreversible inhibition mechanism.