Insights from the yield, protein production, and detailed alkaloid composition of white (Lupinus albus), narrow-leafed (Lupinus angustifolius), and yellow (Lupinus luteus) lupin cultivars in the Mediterranean region.

Introduction: Lupins and other legumes have been considered as alternative plant-based protein sources to soybeans for both humans and livestock. Furthermore, they can contribute to more sustainable agricultural systems. The productivity and chemical composition of legumes is highly variable between species, cultivars, and with the edaphoclimatic conditions. Methods: This work evaluated the adaptability of seven Lupinus cultivars in two different sowing locations, during two consecutive years, through the characterization of their seed, as a means of investigating their suitability to be used as a source of food and/or feed. Results and discussion: Lupinus angustifolius cv. Tango and Lupinus luteus cv. Acos were the most stable genotypes across the environments when considering the seed and protein production, while L. luteus cv. Alburquerque and L. luteus cv. Mister showed less variation in the total alkaloid content across the environments. The edaphoclimatic conditions affected seed and protein yields, as higher rainfall resulted in high productivity. The lower temperatures observed in the first year at both locations caused a reduction in the production of alkaloids in L. luteus cv. Acos and Cardiga. Due to the high alkaloid content of some of the studied cultivars their use as food or feed can pose some safety concerns. However, these cultivars can have high levels of resistance to herbivore and insect attacks, which can be of the utmost importance for the use of these crops for recovering poor or exhausted soils.

Effect of Total Replacement of Soya Bean Meal by Whole Lupine Seeds and of Gender on the Meat Quality and Fatty Acids Profile of Growing Rabbits.

In Europe, the most appropriate strategy to replace soybean meal (SBM) in animal feed has been the development of diets containing locally produced protein sources. One of these sources is lupine (Lupinus spp.). The effect of the total substitution of SBM by white lupine (WL) and yellow lupine (YL) seeds in the diets of growing rabbits and of gender on meat quality and the fatty acids (FA) profile were evaluated. Sixty hybrid weaned rabbits (New Zealand × Californian) (20 rabbits per diet), were fed diets that contained 150 g/kg of SBM (SBMD) and WL (WLD) or YL (YLD) for 35 to 69 days. At the end of this period, 30 rabbits (10 rabbits per diet) were slaughtered to evaluate the carcass and meat characteristics and the FA profile of the longissimus dorsi (LD) muscle. In general, the carcass and meat characteristics (pH and colour) were not affected (p > 0.05) by diet or gender. Further, there was no observed effect (p > 0.05) of gender on meat FA and on the calculated indexes related to human health. However, diet had an effect (p < 0.05) on the FA profile, FA categories, and calculated indexes related to human health. The meat from rabbits fed SBMD presented higher (p < 0.05) saturated FA (SFA; 44 vs. 39 g/100 g average on lupine diets) and lower (p < 0.05) polyunsaturated FA (PUFA; 24 vs. 28 g/100 g average on lupine diets). Our results showed that SBM may be completely replaced by WL or YL, improving the quality of LD muscle FA in terms of nutritional quality for humans.

Introducing Mediterranean Lupins in Lamb Diets: Effects on Carcass Composition, Meat Quality, and Intramuscular Fatty Acid Profile.

The objective of this preliminary study was to evaluate the effects of partial replacement of soybean meal by lupins on lambs' diets, on the carcass traits, meat characteristics, and meat fatty acid profile. Two trials were conducted: In trial 1, the soybean meal (control; C) was partially replaced by Lupinus albus or Lupinus luteus (50 g/kg; LA5 and LL5, respectively); in trial 2, lambs were fed four diets with graded levels of Lupinus luteus (0, 100, 150 and 200 g/kg; C, LL10, LL15, LL20, respectively). At the end of the feeding trials, animals were slaughtered to evaluate carcass characteristics and meat composition, including fatty acids. Carcass composition in tissues was not affected (p > 0.05) by diet in both trials. Additionally, no significant (p < 0.05) differences were observed in meat quality attributes between diets in trials 1 and 2. Overall, the Longissimus muscle's fatty acid content was not affected by diet (p > 0.05) in both trials. Carcass and meat quality was overall comparable between lambs fed with soybean meal and lupins, indicating the latter as a potential alternative protein source. However, the lack of significant differences could also be attributed to the small sample size.

Introducing Mediterranean Lupins in Lambs' Diets: Effects on Growth and Digestibility.

Lupins are suitable candidates to replace soybean meal in livestock feeding in the Mediterranean area, presenting a solution for the European Union's dependence on soybean importations. This study aimed to assess the effect of incorporating Lupinus albus and Lupinus luteus into Churra da Terra Quente lambs' diets on growth performance and digestibility. Two trials were conducted over two years. In trial 1, two experimental diets containing 50 g/kg Lupinus albus and 50 g/kg Lupinus luteus were tested. In trial 2, lambs were fed with diets containing higher incorporations of Lupinus luteus (100, 150, and 200 g/kg: LL10, LL15, and LL20, respectively). Total dry matter, hay dry matter, and crude protein intake were calculated, as well as average daily gains. At the end of the growth trials, dry matter, organic matter, and NDF digestibility was determined. Incorporating 50 g/kg of lupins did not affect (p > 0.05) the performance. Lambs fed on LL20 diets presented the lowest HDMI and CPI values (p < 0.05). The highest intakes (p < 0.05) were observed from LL15 lambs. No differences were found in apparent digestibility coefficients between diets (p > 0.05), except for NDF digestibility which was highest (p < 0.05) for LL20. The optimum level of lupin inclusion in lambs' diets seems to be 150 g/kg.

Zebrafish sex differentiation and gonad development after exposure to 17α-ethinylestradiol, fadrozole and their binary mixture: A stereological study.

Current knowledge on zebrafish (Danio rerio) sex determination suggests that this trait has a polygenic genetic basis, although environmental factors, such as endocrine disrupting chemicals (EDC), may also be involved in modeling or disturbing the species sex differentiation and development. This study aimed to assess how sex steroids imbalance triggers impact on sex differentiation and gonad development in zebrafish. Fish where exposed to an estrogen (EE2, i.e. 17α-ethinylestradiol, 4ng/L), to an inhibitor of estrogen synthesis (Fad, i.e. fadrozole, 50µg/L) or to their binary mixture (Mix-EE2+ Fad, 4ng/L+50µg/L), from 2h to 60 days post-fertilization (dpf). Afterwards, a quantitative (stereological) analysis using light microscopy, based on systematic sampling, was made at 35 and 60dpf, to identify alterations on gonad differentiation and development. During the sex differentiation period, our histological data showed that not all zebrafish males develop a "juvenile ovary", contrarily to what is currently taken for granted. Furthermore, the stereological analysis suggests that EE2 alone enhanced both zebrafish growth and gonad development. On the other hand, exposure to Fad affected the sexual development in zebrafish, inducing masculinization of the specimens, with some degree of intersex observed in males. In addition, the binary mixture allowed identifying sex-dependent roles of steroid hormones in the general growth and gonad development of zebrafish, with estrogens acting as growth promoters in females and being essential for ovary development. Data further support that sex-specific and single EDC impact studies are important, but clearly not sufficient to understand what may occur in the environment.

Effects of Cd injection on osmoregulation and stress indicators in freshwater Nile tilapia.

Freshwater tilapia (Oreochromis niloticus) were intraperitoneally injected with sublethal doses of cadmium (1.25 or 2.5 mg Cd kg(-1) body mass) and sampled after 1, 4 and 7 days in order to evaluate the mechanisms of Cd toxicity at physiological and biochemical levels. Cd levels were significantly elevated in the gill and kidney following injection however levels in the kidney continued to accumulate while levels in the gill either did not change or decreased with time. Cd caused a generalized stress condition as indicated by an increase in blood glucose, lactate and cortisol levels as well as an oxidative stress indicated by increases in lipid peroxidation and protein carbonyl content. Furthermore, tilapia exhibited impairment in their osmoregulatory status based on the fall in plasma sodium levels. Concerning ion regulatory disruption, the kidney was the most affected organ since there was a generalized increase in renal Na(+)/K(+)-ATPase activity after 1 day of exposure to Cd followed by a significant decrease in day 7. This study provides some insights into the mechanisms of Cd toxicity at physiological and biochemical levels and complements previously reported findings on O. niloticus. The disruption of ion homeostasis, alterations in Na(+)/K(+)-ATPase activity and oxidative damage are the effects of Cd exposure that can be integrated in a comprehensive model for Cd impacts.

Metabolic and osmoregulatory changes and cell proliferation in gilthead sea bream (Sparus aurata) exposed to cadmium.

The impact of cadmium on metabolism and osmoregulation was assessed in gilthead sea bream (Sparus aurata). Seawater acclimated fish were injected intraperitoneally with a sublethal dose of cadmium (1.25 mg Cd/kg body wt). After 7 days, half of the injected fish were sampled. The remaining fish were transferred to hypersaline water and sampled 4 days later. Gill and kidney Na(+)/K(+)-ATPase activities, plasma levels of cortisol, several metabolites and osmolytes, as well as osmolality were measured. Hepatosomatic index and condition factor were calculated. The expression levels of Na(+)/K(+)-ATPase, heat shock proteins (HSP70, HSP90) and proliferating cell nuclear antigen was assessed by western blotting. Cadmium treatment adversely affected the Na(+)/K(+)-ATPase activity, although, there was no perturbation in ion homeostasis and the animals were not compromised following transfer to hypersaline water. Increased cell proliferation and Hsp90 expression likely contributed to the attenuation of the deleterious effects of cadmium exposure.

Gill histopathological alterations in Nile Tilapia, Oreochromis niloticus exposed to treated sewage Water

Adult Nile tilapia, Oreochromis niloticus, of both sexes were exposed in wastewater from a sewage treatment plant for a period of 4 days. Gill samples were collected after 24, 48, 72 and 96 h and histopathological changes were analyzed by light and scanning electronic microscopy. Gill epithelium of control O. niloticus (freshwater group) was similar to that of other teleosts, while histopathological lesions were observed in exposed fishes. The main histopathological changes were edema, lifting of lamellar and filamentar epithelia and lamellar fusion. Cell proliferation with consequent thickening of the filament epithelium was also found in fishes exposed to the treated sewage water. The severity of the lesions increased with the time of exposure, namely the hyperplasia of the epithelial cells with proliferation of filamentar epithelium and fusion of lamellae observed at 96 h. Additionally, several histopathological results obtained by light microscopy were confirmed through scanning microscopy.

Tilápias adultas, Oreochromis niloticus, de ambos os sexos foram expostas em águas residuais de uma estação de tratamento de esgoto durante 4 dias. Amostras de brânquia foram recolhidas após 24, 48, 72 e 96 h e as alterações histopatológicas foram analisadas por microscopia óptica e eletrônica de varredura. O epitélio da brânquia do grupo controle apresentou uma morfologia similar à de outros peixes teleosteos, enquanto foram observadas lesões nos peixes expostos. As principais alterações histopatológicas foram edema, destacamento dos epitélios lamelar e filamentar e fusão lamelar. Os peixes expostos às águas residuais mostraram também proliferação celular com consequente aumento da espessura do filamento branquial. A severidade das lesões aumentou com o tempo de exposição, nomeadamente a hiperplasia das células epiteliais com proliferação do epitélio filamentar e fusão das lamelas observadas preferencialmente às 96 h. Adicionalmente, alguns dos resultados obtidos pela análise em microscopia de luz foram confirmados com o recurso da microscopia eletrônica de varredura.

Histopathological changes in liver and gill epithelium of Nile tilapia, Oreochromis niloticus, exposed to waterborne copper

Nile tilapia, Oreochromis niloticus, of both sexes were reared in freshwater and exposed to 0.5, 1.0 and 2.5mg L-1 of waterborne copper for a period of 21 days. Liver and gill samples were collected after 21 days of exposure to copper and lesions were analyzed by light microscopy. The main histopathological changes observed in gills exposed to the highest concentration were edema, lifting of lamellar epithelia and an intense vasodilatation of the lamellar vascular axis. Although less frequent, lamellar fusion caused by the filamentar epithelium proliferation and some lamellar aneurisms were also found. The liver of control group exhibited a quite normal architecture, while the fish exposed to copper showed vacuolation and necrosis. These hepatic alterations were more evident in fish exposed to 1.0 and 2.5mg L-1 copper concentrations. The number of hepatocytes nucleus per mm² of hepatic tissue decreased with the increase of copper concentration. In contrast, the hepatic somatic index was high in fish exposed at 2.5mg L-1 of copper. In short, this work advance new knowledge as influence of copper in the gill and liver histology of O. niloticus and demonstrated that their effects could be observed at different concentrations.

Tilápia do Nilo, Oreochromis niloticus, de ambos os sexos foram mantidas em água doce e expostas a concentrações de 0.5, 1.0 e 2.5mg L-1 de sulfato de cobre durante um período de 21 dias. Amostras de fígado e de brânquia foram coletadas após o tempo de exposição e as lesões foram analisadas por microscopia óptica. As principais alterações histopatológicas observadas nas brânquias foram edema, "lifting" do epitélio lamelar e uma intensa vasodilatação do eixo lamelar. Embora menos freqüentes, a fusão lamelar causada pela proliferação do epitélio filamentar e alguns aneurismas lamelares também foram observadas. O fígado do grupo controle exibiu uma arquitetura normal, enquanto os dos peixes expostos ao cobre apresentaram vacuolização e necrose. Estas lesões hepáticas foram mais evidentes nos peixes expostos a concentrações de cobre de 1.0 e 2.5mg L-1. O número dos núcleos dos hepatócitos / mm² de tecido hepático decresceu com o aumento da concentração de cobre. Em contraste, o índice hepatossomático foi mais elevado nos peixes expostos a 2.5mg L-1 de cobre. Em resumo, este trabalho acrescenta nova informação sobre a influência do cobre ao nível da histologia da brânquia e do fígado de O. niloticus e demonstra que os seus efeitos podem ser observados em diferentes concentrações.

Cadmium tolerance in the Nile tilapia (Oreochromis niloticus) following acute exposure: assessment of some ionoregulatory parameters.

The Nile tilapia (Oreochromis niloticus) can tolerate very high levels of waterborne cadmium. It has one of the highest 96 h LC50 recorded for a freshwater teleost fish (14.8 mg/L Cd; hardness 50 mg/L CaCO(3)). Cadmium is known to perturb ion balance in teleost fishes. However, in an acute time course experiment, plasma Na(+) concentrations were unaffected, and plasma Ca(2+) values only decreased after 96 h exposure in a dose-independent manner. Branchial Na(+)/K(+)-ATPase activity and alpha-subunit protein level expression in crude gill homogenates were not affected by Cd exposure during this period. Branchial chloride cell numbers, identified as Na(+)/K(+)-ATPase immunoreactive cells using immunohistochemistry, decreased 24 h after exposure but recovered thereafter. Histopathological changes did not follow a consistent pattern of variation with exposure time, and the alterations noted in gill epithelium were basically nonspecific to cadmium. Because of its tolerance, it can be concluded that the tilapia O. niloticus would not be a suitable test organism to evaluate sublethal toxicity of cadmium and the realistic impact of this pollutant in the environment. However, it certainly could contribute significantly to our understanding of the toxic mechanism of cadmium exposure in aquatic organisms. This is the first work to investigate the effect of waterborne pollutants on Na(+)/K(+)-ATPase alpha-subunit protein expression in fish gills.