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An amendment to this paper has been published and can be accessed via a link at the top of the paper.

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Understanding the effect of pesticides on the survival of honeybee colonies is important because these pollinators are reportedly declining globally. In the present study, we examined the changes in the head proteome of nurse honeybees exposed to individual and combined pesticides (the fungicide pyraclostrobin and the insecticide fipronil) at field-relevant doses (850 and 2.5 ppb, respectively). The head proteomes of bees exposed to pesticides were compared with those of bees that were not exposed, and proteins with differences in expression were identified by mass spectrometry. The exposure of nurse bees to pesticides reduced the expression of four of the major royal jelly proteins (MRJP1, MRJP2, MRJP4, and MRJP5) and also several proteins associated with carbohydrate metabolism and energy synthesis, the antioxidant system, detoxification, biosynthesis, amino acid metabolism, transcription and translation, protein folding and binding, olfaction, and learning and memory. Overall, when pyraclostrobin and fipronil were combined, the changes in protein expression were exacerbated. Our results demonstrate that vital proteins and metabolic processes are impaired in nurse honeybees exposed to pesticides in doses close to those experienced by these insects in the field, increasing their susceptibility to stressors and affecting the nutrition and maintenance of both managed and natural colonies.

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This study aimed to evaluate the quality of the royal jelly produced by Apis mellifera bees in the presence of different iron concentrations (ferrous sulfate heptahydrate-0, 25, 50, and 100 mg L-1). Two-dimensional electrophoresis was used for the fractionation of royal jelly proteins, and iron level was quantified using flame atomic absorption spectrometry technique. The proteins were identified using electrospray ionisation mass spectrometry. Analysis of variance followed by the Tukey test (P < 0.05) was utilised. Dietary supplementation with mineral Fe affected the protein content and number of proteins in the experimental period. Further, the diet containing the highest iron concentration showed a greater number of spots containing iron, as well as in the abdomen of the bees. The most protein containing Fe were classified as major royal jelly proteins. These results showed that Fe influenced the quality of royal jelly and can improve its nutritional value.

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Fish is an important source of protein, vitamins, and minerals. However, this food is also a major source of human exposure to toxic contaminants such as mercury. Thus, this paper aimed to evaluate mercury-binding proteins for possible application as biomarkers of mercury contamination in hepatic and renal tissues of Plagioscion squamosissimus (carnivorous fish) and Colossoma macropomum (omnivorous fish) from the Amazon region using metalloproteomic approach. The proteome of hepatic and renal tissues of fish species was separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), and the mercury concentrations in protein spots were determined by graphite furnace atomic absorption spectrometry (GFAAS). Finally, the protein spots associated to mercury were characterized by electrospray ionization mass spectrometry (ESI-MS/MS). The activity of antioxidant enzymes (SOD, CAT, GPx, and GST) and lipid peroxidation (LPO) were also determined. The results showed that the highest concentrations of mercury were found in the carnivorous species (P. squamosissimus) and that the accumulation pattern of this metal was higher in hepatic tissues than in renal tissues for both species. A tendency was observed for greater enzymatic activity in the hepatic and renal tissues of P. squamosissimus, the species with the highest concentration of mercury. Only GPx activity in the kidney and GST in the liver were lower for the P. squamosissimus species, and this finding can be explained by the interaction of mercury with these enzymes. The data obtained by ESI-MS/MS allowed for the characterization of the protein spots associated to mercury, revealing proteins involved in energy metabolism, biomolecules transport, protein synthesis and degradation, cell differentiation, gene regulation, and the antioxidant system. The results obtained in the present study can contribute to understanding the physiological processes underlying mercury toxicity and have provided new perspectives on possible candidates for mercury contamination biomarkers in fish.

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High concentrations of mercury found in soils, sediments, fish, and humans of the Amazon region have gained prominence in scientific studies during the last decade. However, studies related to the elucidation of mercury toxicity mechanisms in ichthyofauna at the molecular and metallomic levels that seek to elucidate physiological and functional aspects, as well as the search for biomarkers of mercury exposure, are still sparse. In the search for these answers, the present study analyzed the hepatic tissue proteome of the Arapaima gigas (pirarucu) fish species collected in the Jirau hydroelectric power plant reservoir in the state of Rondônia state, Brazil, in order to identify mercury-related metal-binding proteins and to elucidate their physiological and functional aspects. The proteomic profile of the hepatic tissue of Arapaima gigas was obtained by two-dimensional electrophoresis (2D-PAGE) and the presence of mercury was mapped in the protein SPOTS by graphite furnace atomic absorption spectrometry(GFAAS). Mercury was detected in 18 protein SPOTS with concentrations ranging from 0.13 ±â€¯0.003 to 131.00 ±â€¯3 mg kg-1. The characterization of the protein SPOTS associated with mercury was performed by electrospray ionisation tandem mass spectrometry (ESI-MS/MS), and 10 proteins were identified. Bioinformatics analyses showed that most of the proteins found linked to mercury were involved in cellular component processes and biological processes. For the most part, protein sequences have cellular functions comprising catalytic, binding, sense of localization, and metabolic processes.

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This paper presents a slurry sampling method for total mercury determination by graphite furnace atomic absorption spectrometry (GFAAS) in tissue of fish from the Amazon. The tissue samples were lyophilized and macerated, and then the slurry samples were prepared by putting 20 mg of tissue, added to a solution containing Triton X-100, Suprapur HNO3, and zirconium nitrate directly in sampling vials of a spectrometer. Mercury standard solutions were prepared under the same conditions as the slurry samples. The slurry samples and the mercury standard solutions were sonicated for 20 s. Twenty microliters of slurry samples were injected into the graphite tube, which contained an internal wall lined with tungsten carbide. Under these conditions, it was possible to thermally stabilize the mercury up to an atomization temperature of 1700 °C. The method was validated by mercury determination in reference materials DORM-4 and DOLT-4. The LOD and LOQ were 0.014 and 0.045 mg kg-1, respectively, and recovery percentages in relation to the concentration values were certified in the order of 98%.

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The aim of the present study was to determine the lysine requirement of common carp fingerlings. Two hundred and eighty common carp fingerlings, with mean weight and mean length initial of 0.62 ± 0.05 g and 3.38 ± 0.11 cm, respectively were randomly distributed into 28 squares net cages of 150 litres arranged inside a masonry circular tank with water capacity of 25 m-3 (seven treatments and four replications). The fish were fed until apparent satiation, four times per day, with isoproteic and isoenergetic diets supplemented with lysine at 1.58, 1.81, 2.05, 2.29, 2.53, 2.76, and 3.0% of the diet for 57 days. Productive performance, survival rate, and chemical composition of the fish body were evaluated at the end of the study. The data obtained was subjected to regression analysis and, when significantly different (P<0.05), the Tukey multiple comparison test was used. Additionally, linear response plateau (LRP) analysis was performed. Lysine supplementation significantly improved (P<0.05) mean final weight, mean weight gain, and specific growth rate. However, lysine levels above 2.29% compromised the feed efficiency. Lysine requirements of common carp fingerlings were found to be 1.89% of the diet or 4.73% of the crude protein in the diet for obtain better growth.(AU)

O presente estudo teve por objetivo determinar a exigência de lisina para alevinos de carpa comum (Cyprinus carpio). Foram distribuídos inteiramente ao acaso 280 alevinos de carpa comum com peso e comprimento inicial médio de 0,62 ± 0,05 g e 3,38 ± 0,11 cm, respectivamente, em 28 tanques-rede quadrados com volume útil de 150 litros, dispostos no interior de um tanque circular de alvenaria com capacidade para 25 m3 de água (sete tratamentos e quatro repetições). Os peixes foram alimentados até a saciedade aparente quatro vezes ao dia com dietas isproteicas e isoenergéticas suplementadas com lisina nas concentrações de 1,58; 1,81; 2,05; 2,29; 2,53; 2,76 e 3,0% da dieta por 57 dias. Ao final do estudo foi avaliado o desempenho produtivo, sobrevivência e composição centesimal da carcaça dos peixes. Os dados obtidos foram submetidos a análise de regressão (p<0,05) e ao teste múltiplo de média Tukey. A análise do platô de resposta linear (LRP) foi aplicado adicionalmente. A suplementação de lisina melhorou (p<0,05) o peso final médio, ganho de peso médio e taxa de crescimento específico e a partir de 2,29% de suplementação de lisina na dieta prejudicou a eficiência alimentar dos peixes. A exigência de lisina para alevinos de carpa comum é de 1,89% da dieta ou 4,73% da proteína bruta para obtenção do melhor crescimento.(AU)

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The aim of the present study was to determine the lysine requirement of common carp fingerlings. Two hundred and eighty common carp fingerlings, with mean weight and mean length initial of 0.62 ± 0.05 g and 3.38 ± 0.11 cm, respectively were randomly distributed into 28 squares net cages of 150 litres arranged inside a masonry circular tank with water capacity of 25 m-3 (seven treatments and four replications). The fish were fed until apparent satiation, four times per day, with isoproteic and isoenergetic diets supplemented with lysine at 1.58, 1.81, 2.05, 2.29, 2.53, 2.76, and 3.0% of the diet for 57 days. Productive performance, survival rate, and chemical composition of the fish body were evaluated at the end of the study. The data obtained was subjected to regression analysis and, when significantly different (P<0.05), the Tukey multiple comparison test was used. Additionally, linear response plateau (LRP) analysis was performed. Lysine supplementation significantly improved (P<0.05) mean final weight, mean weight gain, and specific growth rate. However, lysine levels above 2.29% compromised the feed efficiency. Lysine requirements of common carp fingerlings were found to be 1.89% of the diet or 4.73% of the crude protein in the diet for obtain better growth.

O presente estudo teve por objetivo determinar a exigência de lisina para alevinos de carpa comum (Cyprinus carpio). Foram distribuídos inteiramente ao acaso 280 alevinos de carpa comum com peso e comprimento inicial médio de 0,62 ± 0,05 g e 3,38 ± 0,11 cm, respectivamente, em 28 tanques-rede quadrados com volume útil de 150 litros, dispostos no interior de um tanque circular de alvenaria com capacidade para 25 m3 de água (sete tratamentos e quatro repetições). Os peixes foram alimentados até a saciedade aparente quatro vezes ao dia com dietas isproteicas e isoenergéticas suplementadas com lisina nas concentrações de 1,58; 1,81; 2,05; 2,29; 2,53; 2,76 e 3,0% da dieta por 57 dias. Ao final do estudo foi avaliado o desempenho produtivo, sobrevivência e composição centesimal da carcaça dos peixes. Os dados obtidos foram submetidos a análise de regressão (p<0,05) e ao teste múltiplo de média Tukey. A análise do platô de resposta linear (LRP) foi aplicado adicionalmente. A suplementação de lisina melhorou (p<0,05) o peso final médio, ganho de peso médio e taxa de crescimento específico e a partir de 2,29% de suplementação de lisina na dieta prejudicou a eficiência alimentar dos peixes. A exigência de lisina para alevinos de carpa comum é de 1,89% da dieta ou 4,73% da proteína bruta para obtenção do melhor crescimento.

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O presente trabalho avaliou a influência da adição de urucum como agente pigmentante sobre as características de coloração da pele, músculo e desempenho produtivo de Carassius auratus, criados em hapas de 0,15 m3 (dimensões de 0,50 x 0,50 x 0,65 m), dispostos no interior de um tanque de alvenaria circular de 25 m3. Foram utilizados 336 alevinos de C. auratus distribuídos inteiramente ao acaso em 28 hapas, com sete tratamentos e quatro repetições, com 12 peixes por unidade. Os níveis de adição de urucum foram 0,0; 0,25; 0,50; 1,00; 2,00; 4,00 e 8,00%. Os peixes com peso inicial médio de 1,12 ± 0,18 g e comprimento de 4,31 ± 0,44 cm, foram alimentados as 08, 11, 14 e 17 horas. A característica colorimétrica de b* (amarelo) mostrou valores significativos aos 96 e 141 dias no sistemade coordenadas Hunter com adição de 2,0 e 1,0% de urucum na dieta, respectivamente e, no sistema de coordenadas CMYK aos 96 dias para C (ciano) e aos 141 dias para Y (amarelo) com adição de 8,0 e 2,0% de urucum na dieta respectivamente. Observaram-se diferenças (P<0,05) para peso final médio, ganho em peso médio, conversão alimentar aparente, taxa de crescimento diário e eficiência alimentar. Recomenda-se uma dieta contendo 1,0% de adição de urucum para C. auratus para o melhor desempenho e de 1,0 a 4,0% de adição de urucum para intensificação luminosa da pele dos peixes.(AU)

This study evaluated the effect of adding annatto as agent pigmentation on the staining characteristics of the skin, muscle and productive performance of Carassius auratus, reared in 0.15 m3 (dimensions 0.50 x 0.50 x 0.65 m) hapas disposed within a circular masonry tank of 25 m3. We used 336 C. auratus fingerlings completely randomized design in 28 hapas with seven treatments and four replicates with 12fish per unit. The addition of annatto levels were 0.0, 0.25, 0.50, 1.00, 2.00, 4.00 and 8.00%. Fish with an average initial weight of 1.12 ± 0.18 g and length of 4.31 ± 0.44 cm, were fed the 08, 11, 14 and 17 hours. The b * colorimetric characteristics (yellow) showed significant at 96 and 141 days in the Hunter coordinate system with the addition of 2.0 and 1.0% annatto in the diet, respectively, and the coordinatesystem of CMYK and 96 days for C (cyan), and 141 days for the Y (yellow), with the addition of 8.0 to 2.0% annatto in the diet, respectively. We observed differences (P <0.05) average final weight, weight gain, feed conversion, daily growth rate and feed efficiency. It is recommended a diet containing 1.0% added annatto to C. auratus to better performance and 1.0 to 4.0% added annatto to intensified lightfish skin.(AU)

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Vitamin A is important to the proper development and functioning of the body, appearing in several metabolic functions. This experiment was conducted with aim evaluate to the vitamin A supplementation in diets for Goldfish Carassius auratus. The experimental was completely randomized with six treatments (0, 2.000, 4.000, 8.000, 16.000 and 32.000 IU of vitamin A kg-1 diet) with four repetitions into 24 experimental cages, with capacity for 150 liters useful volume, made of mesh shading, installed inside a masonry circular tank with a capacity of 25 m3 of water. 192 fish with an average initial weight of 6.66±0.57 g and initial length of 7.30±0.33 cm, feed at 8h, 11h, 14h and 17h at satiety were used. Evaluated the effects on productive performance and intensity of yellow-red coloring of the skin of the fish were. The Linear Response Plateau analysis of weight gain and length total, feed conversion and daily growth rate versus vitamin A requirement estimate showed 2,624, 2,385 and 2,358, 4,381 and 2,866 IU of vitamin A per kg diet, respectively. The data indicated that this vitamin has little or no influence on the intensity of the skin color of the Goldfish.(AU)

A vitamina A é importante para o correto desenvolvimento e funcionamento do organismo, figurando em diversas funções metabólicas. Foi conduzido um experimento com o objetivo de avaliar a suplementação de vitamina A em dietas para kinguio Carassius auratus. O delineamento experimental foi inteiramente casualizado com seis tratamentos e quatro repetições (0, 2.000, 4.000, 8.000, 16.000 e 32.000 UI de vitamina A kg-1 de dieta) em 24 tanques rede experimentais, com capacidade para 150 litros de volume útil, confeccionados em malha sombrite, instalados no interior de um tanque circular de alvenaria com capacidade para 25 m3 de água. Foram utilizados 192 peixes com peso inicial médio de 6,66±0,57g e comprimento inicial médio de 7,30±0,33 cm, alimentados às 8h, 11h, 14h e 17h ate a saciedade. Foram avaliados os efeitos sobre o desempenho produtivo e intensidade de coloração vermelho-amarelada da pele dos peixes. A análise de Linear Response Plateau do ganho de peso, comprimento padrão e total, conversão alimentar aparente e taxa de crescimento diário versus níveis de vitamina A demonstrou estimativa de exigência de 2.624, 2.385 e 2.358, 4.381 e 2.866 UI de vitamina A por kg de ração, respectivamente. Os dados indicam que aparentemente esta vitamina tem pouca ou nenhuma influência sobre a intensidade da cor da pele do kinguio.(AU)

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Vitamin A is important to the proper development and functioning of the body, appearing in several metabolic functions. This experiment was conducted with aim evaluate to the vitamin A supplementation in diets for Goldfish Carassius auratus. The experimental was completely randomized with six treatments (0, 2.000, 4.000, 8.000, 16.000 and 32.000 IU of vitamin A kg-1 diet) with four repetitions into 24 experimental cages, with capacity for 150 liters useful volume, made of mesh shading, installed inside a masonry circular tank with a capacity of 25 m3 of water. 192 fish with an average initial weight of 6.66±0.57 g and initial length of 7.30±0.33 cm, feed at 8h, 11h, 14h and 17h at satiety were used. Evaluated the effects on productive performance and intensity of yellow-red coloring of the skin of the fish were. The Linear Response Plateau analysis of weight gain and length total, feed conversion and daily growth rate versus vitamin A requirement estimate showed 2,624, 2,385 and 2,358, 4,381 and 2,866 IU of vitamin A per kg diet, respectively. The data indicated that this vitamin has little or no influence on the intensity of the skin color of the Goldfish.

A vitamina A é importante para o correto desenvolvimento e funcionamento do organismo, figurando em diversas funções metabólicas. Foi conduzido um experimento com o objetivo de avaliar a suplementação de vitamina A em dietas para kinguio Carassius auratus. O delineamento experimental foi inteiramente casualizado com seis tratamentos e quatro repetições (0, 2.000, 4.000, 8.000, 16.000 e 32.000 UI de vitamina A kg-1 de dieta) em 24 tanques rede experimentais, com capacidade para 150 litros de volume útil, confeccionados em malha sombrite, instalados no interior de um tanque circular de alvenaria com capacidade para 25 m3 de água. Foram utilizados 192 peixes com peso inicial médio de 6,66±0,57g e comprimento inicial médio de 7,30±0,33 cm, alimentados às 8h, 11h, 14h e 17h ate a saciedade. Foram avaliados os efeitos sobre o desempenho produtivo e intensidade de coloração vermelho-amarelada da pele dos peixes. A análise de Linear Response Plateau do ganho de peso, comprimento padrão e total, conversão alimentar aparente e taxa de crescimento diário versus níveis de vitamina A demonstrou estimativa de exigência de 2.624, 2.385 e 2.358, 4.381 e 2.866 UI de vitamina A por kg de ração, respectivamente. Os dados indicam que aparentemente esta vitamina tem pouca ou nenhuma influência sobre a intensidade da cor da pele do kinguio.

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O presente trabalho avaliou a influência da adição de urucum como agente pigmentante sobre as características de coloração da pele, músculo e desempenho produtivo de Carassius auratus, criados em hapas de 0,15 m3 (dimensões de 0,50 x 0,50 x 0,65 m), dispostos no interior de um tanque de alvenaria circular de 25 m3. Foram utilizados 336 alevinos de C. auratus distribuídos inteiramente ao acaso em 28 hapas, com sete tratamentos e quatro repetições, com 12 peixes por unidade. Os níveis de adição de urucum foram 0,0; 0,25; 0,50; 1,00; 2,00; 4,00 e 8,00%. Os peixes com peso inicial médio de 1,12 ± 0,18 g e comprimento de 4,31 ± 0,44 cm, foram alimentados as 08, 11, 14 e 17 horas. A característica colorimétrica de b* (amarelo) mostrou valores significativos aos 96 e 141 dias no sistemade coordenadas Hunter com adição de 2,0 e 1,0% de urucum na dieta, respectivamente e, no sistema de coordenadas CMYK aos 96 dias para C (ciano) e aos 141 dias para Y (amarelo) com adição de 8,0 e 2,0% de urucum na dieta respectivamente. Observaram-se diferenças (P<0,05) para peso final médio, ganho em peso médio, conversão alimentar aparente, taxa de crescimento diário e eficiência alimentar. Recomenda-se uma dieta contendo 1,0% de adição de urucum para C. auratus para o melhor desempenho e de 1,0 a 4,0% de adição de urucum para intensificação luminosa da pele dos peixes.

This study evaluated the effect of adding annatto as agent pigmentation on the staining characteristics of the skin, muscle and productive performance of Carassius auratus, reared in 0.15 m3 (dimensions 0.50 x 0.50 x 0.65 m) hapas disposed within a circular masonry tank of 25 m3. We used 336 C. auratus fingerlings completely randomized design in 28 hapas with seven treatments and four replicates with 12fish per unit. The addition of annatto levels were 0.0, 0.25, 0.50, 1.00, 2.00, 4.00 and 8.00%. Fish with an average initial weight of 1.12 ± 0.18 g and length of 4.31 ± 0.44 cm, were fed the 08, 11, 14 and 17 hours. The b * colorimetric characteristics (yellow) showed significant at 96 and 141 days in the Hunter coordinate system with the addition of 2.0 and 1.0% annatto in the diet, respectively, and the coordinatesystem of CMYK and 96 days for C (cyan), and 141 days for the Y (yellow), with the addition of 8.0 to 2.0% annatto in the diet, respectively. We observed differences (P <0.05) average final weight, weight gain, feed conversion, daily growth rate and feed efficiency. It is recommended a diet containing 1.0% added annatto to C. auratus to better performance and 1.0 to 4.0% added annatto to intensified lightfish skin.

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