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Comparative Fecal Metabolomes of Silkworms Being Fed Mulberry Leaf and Artificial Diet.

ABSTRACT: Metabonomics accurately monitors the precise metabolic responses to various dietary patterns. Metabolic profiling allows simultaneous measurement of various fecal metabolites whose concentrations may be affected by food intake. In this study, we analyzed the fecal metabolomes of silkworm (Bombyx mori) larvae reared on fresh mulberry leaves and artificial diets. 57 differentially expressed metabolites were identified by gas chromatography-mass spectrometry. Of these, 39 were up-regulated and 18 were downregulated in the mulberry leaf meal group. Most of the amino acids, carbohydrates and lipids associated with physical development and silk protein biosynthesis were enriched in silkworms reared on mulberry leaves. In contrast, the urea, citric acid, D-pinitol, D-(+)-cellobiose and N-acetyl glucosamine levels were relatively higher in the silkworm feeding on the artificial diets. The findings of this study help clarify the association between diet and metabolic profiling.

SUBMITTER: Qin D

PROVIDER: S-EPMC7759890 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature

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Comparative Fecal Metabolomes of Silkworms Being Fed Mulberry Leaf and Artificial Diet.

Qin DaoYuan D Wang GenHong G Dong ZhaoMing Z Xia QingYou Q Zhao Ping P

Insects 20201130 12

Metabonomics accurately monitors the precise metabolic responses to various dietary patterns. Metabolic profiling allows simultaneous measurement of various fecal metabolites whose concentrations may be affected by food intake. In this study, we analyzed the fecal metabolomes of silkworm (<i>Bombyx mori</i>) larvae reared on fresh mulberry leaves and artificial diets. 57 differentially expressed metabolites were identified by gas chromatography-mass spectrometry. Of these, 39 were up-regulated a ...[more]

PMID: 33266201

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Project description:BackgroundSilkworm pupae is the main by-product of the sericulture industry with an interesting nutritional profile, especially in terms of proteins. In consideration of its possible use as a food or food ingredient in Western countries, a comparative proteomic experiment has been performed to investigate the differences of the protein profile of male and female silkworm pupae reared on mulberry leaves or on an artificial diet.MethodsThe nutritional profile of lyophilized silkworm pupae in terms of dry matter and ash was evaluated according to the AOAC procedures, the total nitrogen content was determined by a nitrogen analyzer and the silkworm pupae gross energy value was measured using an adiabatic calorimetric bomb. The comparative proteomic analysis was performed on male and female silkworm pupae reared on mulberry leaves or on the artificial diet. Proteins were separated by two-dimensional electrophoresis and, after a multivariate statistical analysis, the differentially expressed proteins were identified by LC-MS/MS.ResultsThe comparative proteomic approach highlighted 47 silkworm pupae proteins differentially expressed comparing diet and gender. PCA analysis showed that seven proteins were more effective in discriminating the sex and five were more effective in discriminating the diet type. In spite of the above-mentioned differences in the silkworm pupae protein profile, no strong alteration of the pupa physiological traits have been demonstrated, suggesting a general silkworm pupae flexibility to adapt to a well-balanced artificial diet. Differences in lipid transport and metabolism were found among the experimental groups, that might have a relevant effect on the timing and on hormone secretion. This aspect may also affect silk production, as univoltine strains are the most productive. The proteomic data provided in this work, may offer a contribution in understanding also the influence of gender and farming strategy on the allergen profile of Bombyx mori, when used as food or as a food ingredient. Female silkworm pupae reared on mulberry leaves seemed to contain lower levels of known allergens than those reared in the other experimental conditions; these findings will have to be taken into account when farming B. mori for food production purposes. However, our results need to be supported by further characterization of the allergenic potential of B. mori.

Project description:Use of formula feed (FF) for silkworms for all instars, has promoted transformation and progress in traditional sericulture. However, the cocoon yield of FF silkworms has failed to reach that of silkworms fed mulberry leaves (ML). The biological mechanisms underlying this phenomenon have not been well described. This study aimed to identify metabolic mechanisms and potential biomarkers relating to the poor cocoon yield of FF silkworms. In this study, silkworms received treatments of either ML (ML group) or FF (FF group) for all instars. At the 3rd day of the 5th instar, the midgut (MG), hemolymph (HL) and posterior silk gland (PSG) were collected for the metabolome profiles detection. The remaining silkworms were fed ML or FF until cocooning for investigation. The whole cocoon yield (WCY) was significantly higher in the FF group than the ML group (p < 0.05), whereas the cocoon shell weight (CSW) and cocoon shell rate (CSR) were significantly lower in the FF group (p < 0.05). A total of 845, 867 and 831 metabolites were qualified and quantified in the MG, HL and PSG of the FF silkworms, respectively. Correspondingly, 789, 833 and 730 metabolites were quantified in above three tissues of the ML group. Further, 230, 249 and 304 significantly different metabolites (SDMs) were identified in the MG, HL and PSG between the FF and ML group, respectively. Eleven metabolic pathways enriched by the SDMs were mutual among the three tissues. Among them, cysteine and methionine metabolism, arginine biosynthesis, and arginine and proline metabolism were the top three pathways with the highest impact value in the PSG. Six biomarkers were obtained through biomarker analysis and Pearson correlation calculation. Among them, homocitrulline, glycitein, valyl-threonine, propyl gallate and 3-amino-2,3-dihydrobenzoic acid were positively correlated with WCY, but negatively correlated with CSW and CSR (p < 0.05). An opposite correlation pattern was observed between 3-dimethylallyl-4-hydroxyphenylpyruvate and the three cocoon performance traits. Overall, three key metabolic pathways and six biomarkers associated with cocoon yield were interpreted, and should provide directions for formula feed optimization in factory-raised silkworms.

Project description:Background. Silkworm pupae (SWP) is the main by-product of the sericulture industry with an interesting nutritional profile, especially in terms of proteins. In consideration of its possible use as a food or food ingredient in Western countries, a comparative proteomic experiment has been performed to investigate the differences of the protein profile of male and female SWP reared on mulberry leaves or on an artificial diet. Methods. The nutritional profile of lyophilized SWP in terms of dry matter and ash was evaluated according to the AOAC procedures, the total nitrogen content was determined by a nitrogen analyzer and the SWP gross energy value was measured using an adiabatic calorimetric bomb. The comparative proteomic analysis was performed on male and female SWP reared on mulberry leaves or on the artificial diet. Proteins were separated by Bidimensional Electrophoresis (2DE) and, after a multivariate statistical analysis, the differentially expressed proteins were identified by LC-MS/MS. Results. The comparative proteomic approach highlighted 47 SWP proteins differentially expressed comparing diet and gender. PCA analysis showed that 7 proteins were more effective in discriminating the sex and 5 were more effective in discriminating the diet type. In spite of the above mentioned differences in the SWP protein profile, no strong alteration of the pupa physiological traits have been demonstrated, suggesting a general SWP flexibility to adapt to a well-balanced artificial diet. Differences in lipid transport and metabolism were found among the experimental groups, that might have a relevant effect on the timing, on hormone secretion and, in turn, on insect voltinism. This aspect may also affect silk production, as univoltine strains are the most productive. Although this is a preliminary study, the proteomic data may offer a contribution in understanding also the influence of gender and farming strategy on the allergen profile of B. mori, when used as food or as a food ingredient. Since female silkworm pupae reared on mulberry leaves seemed to contain lower levels of known allergens than those reared in the other experimental conditions, we speculated that these findings may help when farming B. mori for food production purposes. However, these results need to be supported by further characterization of the allergenic potential of B. mori.

Dataset Information

Comparative Fecal Metabolomes of Silkworms Being Fed Mulberry Leaf and Artificial Diet.

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Comparative Fecal Metabolomes of Silkworms Being Fed Mulberry Leaf and Artificial Diet.

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