{"id":131,"date":"2019-06-28T21:56:36","date_gmt":"2019-06-28T19:56:36","guid":{"rendered":"http:\/\/espanol.beefree.es\/?p=131"},"modified":"2025-06-25T21:34:31","modified_gmt":"2025-06-25T19:34:31","slug":"jarabe-de-azucar-acorta-la-vida-de-las-abejas","status":"publish","type":"post","link":"https:\/\/espanol.resistantbees.es\/?p=131","title":{"rendered":"Jarabe de az\u00facar acorta la vida de las abejas &#8211; estudio cient\u00edfico"},"content":{"rendered":"<p>[vc_row][vc_column][vc_column_text]<strong>Es un h\u00e1bito com\u00fan de la mayor\u00eda de los apicultores de sacar toda la miel de las colmenas y luego alimentarlas con jarabe de az\u00facar, con lo que pueden pasar el invierno.<\/strong><\/p>\n<p><strong>Pero ahora la pregunta es \u00bfhasta qu\u00e9 punto perjudica esta dieta artificial a las abejas?<\/strong><\/p>\n<div id=\"gt-res-content\">\n<div dir=\"ltr\">Ya hemos demostrado que las abejas de celdillas peque\u00f1as tienen<a href=\"\/?p=427\"> una esperanza de vida m\u00e1s larga que las abejas\u00a0\u00a0<\/a><a href=\"\/?p=427\">artificialmente\u00a0<\/a>agrandadas.<\/div>\n<\/div>\n<p>Y s\u00f3lo a trav\u00e9s de esta vida m\u00e1s larga se estimula el\u00a0<a href=\"\/?p=136\">comportamiento higi\u00e9nico.<\/a><\/p>\n<p><span style=\"font-size: 14pt;\">Ahora, sin embargo, llega un\u00a0<a href=\"\/?p=399\">estudio cient\u00edfico<\/a>\u00a0y dice lo que siempre hemos proclamados:<\/span><br \/>\n<span style=\"font-size: 14pt;\"><a href=\"\/?p=399\"><strong>La alimentaci\u00f3n artificial acorta la vida de las abejas<\/strong><\/a><\/span><\/p>\n<p>Abejas se alimentaron con miel y con diferentes jarabes de repuesto:<\/p>\n<p><img decoding=\"async\" src=\"http:\/\/www.resistantbees.com\/fotos\/estudio\/feeding2.jpeg\" alt=\"\" width=\"550\" \/><\/p>\n<p><strong>Alimentadas con miel estas abejas viv\u00edan un promedio de 27 d\u00edas, con jarabe de az\u00facar s\u00f3lo 21 d\u00edas, y con jarabe \u00e1cido invertido s\u00f3lo 12 d\u00edas, mucho menos de la mitad de la vida que con la miel natural.<\/strong><\/p>\n<p>Y ese fue el resultado resumido:<br \/>\n<img decoding=\"async\" src=\"http:\/\/www.resistantbees.com\/fotos\/estudio\/feeding1.jpeg\" alt=\"\" width=\"550\" \/><\/p>\n<p>en resumen:<br \/>\nSe puede decir que las diversas sustancias de los alimentos tienen diferentes influencias principalmente en la pared intestinal de abejas. Miel natural no tuvo efectos sobre la mucosa intestinal. Mientras que la adici\u00f3n de levadura y malta al jarabe ha conducido a da\u00f1os de la mucosa .<\/p>\n<p>\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014-<a name=\"zweite\"><\/a><\/p>\n<p>Otro estudio muestra:<\/p>\n<p>La alimentaci\u00f3n con az\u00facar influye en la actividad de los genes<\/p>\n<p>&nbsp;<\/p>\n<h3>EE.UU.- LOS CIENT\u00cdFICOS RASTREAN LA ACTIVIDAD DE LOS GENES CUANDO LAS ABEJAS MELLIFERAS COMEN MIEL Y CUANDO NO LO HACEN<\/h3>\n<div>\n<p>Muchos apicultores alimentan a sus abejas melliferas con sacarosa \u00a0o jarabe de ma\u00edz de alta fructosa en aquellos tiempos en que falta el alimento \u00a0dentro de la colmena. Esta pr\u00e1ctica ha sido objeto de escrutinio, sin embargo, en respuesta al desorden del colapso de colonias, la enorme y a\u00fan no totalmente explicada mortandad de las abejas en los EE.UU. y Europa. Algunos sospechan que la nutrici\u00f3n inadecuada juega un papel en la disminuci\u00f3n de la poblaci\u00f3n de las abejas<\/p>\n<\/div>\n<p>&nbsp;<\/p>\n<p>aqui el estudio completo:<\/p>\n<h1><a href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html\">Diet-dependent gene expression in honey bees: honey vs. sucrose or high fructose corn syrup<\/a><\/h1>\n<section>\n<div>\n<div>\n<div id=\"first-paragraph\">\n<p>Severe declines in honey bee populations have made it imperative to understand key factors impacting honey bee health. Of major concern is nutrition, as malnutrition in honey bees is associated with immune system impairment and increased pesticide susceptibility. Beekeepers often feed high fructose corn syrup (HFCS) or sucrose after harvesting honey or during periods of nectar dearth. We report that, relative to honey, chronic feeding of either of these two alternative carbohydrate sources elicited hundreds of differences in gene expression in the fat body, a peripheral nutrient-sensing tissue analogous to vertebrate liver and adipose tissues. These expression differences included genes involved in protein metabolism and oxidation-reduction, including some involved in tyrosine and phenylalanine metabolism. Differences between HFCS and sucrose diets were much more subtle and included a few genes involved in carbohydrate and lipid metabolism. Our results suggest that bees receive nutritional components from honey that are not provided by alternative food sources widely used in apiculture.<\/p>\n<\/div>\n<div><\/div>\n<\/div>\n<\/div>\n<\/section>\n<section>\n<div id=\"introduction\">\n<h1><a>Introduction<\/a><\/h1>\n<div>\n<nav><\/nav>\n<p>Honey bees are vital members of natural and agricultural ecosystems worldwide. In the United States, the Western honey bee (<i>Apis mellifera<\/i>) contributes more than 15 billion dollars to the agricultural industry annually<sup><a id=\"ref-link-1\" title=\"Calderone, N. W. Insect Pollinated Crops, Insect Pollinators and US Agriculture: Trend Analysis of Aggregate Data for the Period 1992\u20132009. PLoS ONE 7, e37235 (2012).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref1\">1<\/a><\/sup>. It is therefore of serious concern that honey bee populations have declined steadily in the United States, with dramatic losses of colonies starting in 2006 associated with colony collapse disorder (CCD)<sup><a id=\"ref-link-2\" title=\"Committee on the Status of Pollinators in North America, National Research Council. [Status of Pollinators in North America]. (The National Academies Press, 2007).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref2\">2<\/a>,\u00a0<a id=\"ref-link-3\" title=\"vanEngelsdorp, D., Hayes, J., Jr, Underwood, R. M. &amp; Pettis, J. A Survey of Honey Bee Colony Losses in the U.S., Fall 2007 to Spring 2008. PLoS ONE 3, e4071 (2008).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref3\">3<\/a>,\u00a0<a id=\"ref-link-4\" title=\"vanEngelsdorp, D. et al. A national survey of managed honey bee 2010-11 winter colony losses in the USA: results from the Bee Informed Partnership. J. of Api. Res. 51, 115\u2013124 (2012).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref4\">4<\/a><\/sup>. These losses have intensified the need to understand factors that impact honey bee health.<\/p>\n<p>Central to honey bee health is nutrition<sup><a id=\"ref-link-5\" title=\"Brodschneider, R. &amp; Crailsheim, K. Nutrition and health in honey bees. Apidologie 41, 278\u2013294 (2010).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref5\">5<\/a><\/sup>. Malnutrition in honey bee colonies can result from maintaining densities of colonies that are too high for available flora or placement of colonies for pollination of crops that are deficient in pollen or nectar or have low nutritive value<sup><a id=\"ref-link-6\" title=\"Brodschneider, R. &amp; Crailsheim, K. Nutrition and health in honey bees. Apidologie 41, 278\u2013294 (2010).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref5\">5<\/a>,\u00a0<a id=\"ref-link-7\" title=\"Haydak, M. H. Honey Bee Nutrition. Annu. Rev. Entomol. 15, 143\u2013156 (1970).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref6\">6<\/a><\/sup>. Poor nutrition can make bees more susceptible to pesticides<sup><a id=\"ref-link-8\" title=\"Wahl, O. &amp; Ulm, K. Influence of pollen feeding and physiological condition on pesticide sensitivity of the honey bee Apis mellifera carnica. Oecologia 59, 106\u2013128 (1983).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref7\">7<\/a><\/sup>\u00a0and lead to a compromised immune system making bees more vulnerable to diseases<sup><a id=\"ref-link-9\" title=\"Alaux, C., Ducloz, F., Crauser, D. &amp; Le Conte, Y. Diet effects on honeybee immunocompetence. Biol. Lett. 6, 562\u2013565 (2010).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref8\">8<\/a><\/sup>.<\/p>\n<p>The principal natural carbohydrate source of honey bees is nectar, which is collected from flowers, transported to the hive and converted to honey for storage. This conversion involves reducing the water content to 16\u201320% and adding glandular secretions that contain microorganisms and enzymes, including amylases, glucose oxidases and invertases<sup><a id=\"ref-link-10\" title=\"Brodschneider, R. &amp; Crailsheim, K. Nutrition and health in honey bees. Apidologie 41, 278\u2013294 (2010).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref5\">5<\/a>,\u00a0<a id=\"ref-link-11\" title=\"Winston, M. L. [The biology of the honey bee]. (Harvard Univ Pr, 1987).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref9\">9<\/a><\/sup>. These increase acidity and convert the sucrose in nectar into glucose and fructose<sup><a id=\"ref-link-12\" title=\"Winston, M. L. [The biology of the honey bee]. (Harvard Univ Pr, 1987).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref9\">9<\/a><\/sup>. The final constituents of honey vary depending on the nectar source but are mainly fructose (30\u201345%), glucose (24\u201340%) and sucrose (0.1\u20134.8%), as well as trace amounts of other disaccharides, vitamins, minerals, amino acids and a variety phenolic compounds<sup><a id=\"ref-link-13\" title=\"White, J. W. The Composition of Honey. Bee World 38, 57\u201366 (1957).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref10\">10<\/a><\/sup>.<\/p>\n<p>Adult honey bees use honey as fuel for energy-intensive flights and colony thermoregulation<sup><a id=\"ref-link-14\" title=\"Winston, M. L. [The biology of the honey bee]. (Harvard Univ Pr, 1987).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref9\">9<\/a><\/sup>. Unlike larvae, adults have low levels of abdominal lipids and cannot survive for long periods of time without a carbohydrate source. A continuous supply of sugar is particularly important for foraging honey bees, because they have a diet that is mainly carbohydrate-based<sup><a id=\"ref-link-15\" title=\"Crailsheim, K. et al. Pollen consumption and utilization in worker honeybees (Apis mellifera carnica): Dependence on individual age and function. J. Insect Physiol. 38, 409\u2013419 (1992).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref11\">11<\/a><\/sup>. Compared to younger bees that specialize on performing tasks inside the hive, foragers also have a higher metabolic rate<sup><a id=\"ref-link-16\" title=\"Harrison, J. M. Caste-specific changes in honeybee flight capacity. Physio.l zool. 59, 175\u2013187 (1986).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref12\">12<\/a><\/sup>\u00a0and lose over half their abdominal lipid stores prior to starting to forage<sup><a id=\"ref-link-17\" title=\"Toth, A. L. &amp; Robinson, G. E. Worker nutrition and division of labour in honeybees. Anim. Behav. 69, 427\u2013435 (2005).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref13\">13<\/a><\/sup>.<\/p>\n<p>Beekeepers often provide supplemental carbohydrates in the form of high fructose corn syrup (HFCS) or sucrose following the harvesting of honey or during periods of nectar dearth. Supplementing with HFCS became a widespread practice following early studies that showed acceptable honey bee survival<sup><a id=\"ref-link-18\" title=\"Barker, R. J. &amp; Lehner, Y. Laboratory comparison of high fructose corn syrup, grape syrup, honey and sucrose syrup as maintenance food for caged honey bees. Apidologie 9, 111\u2013116 (1978).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref14\">14<\/a><\/sup>\u00a0and equivalent honey production and long-term productivity relative to honey feeding<sup><a id=\"ref-link-19\" title=\"Severson, D. W. &amp; Erickson, E. H. Honey bee (Hymenoptera: Apidae) colony performance in relation to supplemental carbohydrates. J. of Econ. Entomol. 77, 1473\u20131478 (1984).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref15\">15<\/a><\/sup>. In addition, HFCS has a fructose-to-glucose ratio similar to honey, with the most common bee feed formulation composed of 55% fructose and 42% glucose<sup><a id=\"ref-link-20\" title=\"Hanover, L. M. &amp; White, J. S. Manufacturing, composition, and applications of fructose. Am. J. Clin. Nutr. 58, 724S\u2013732S (1993).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref16\">16<\/a><\/sup>. HFCS is also less expensive than sucrose and is less labor-intensive to administer as food because it comes in liquid form<sup><a id=\"ref-link-21\" title=\"Barker, R. J. Considerations in selecting sugars for feeding to honey bees. Am. Bee J. 117, 76\u201377 (1977).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref17\">17<\/a><\/sup>.<\/p>\n<p>However, questions regarding the suitability of HFCS for honey bees have arisen, in part because of CCD and because of research showing HFCS may have deleterious metabolic effects in mammals<sup><a id=\"ref-link-22\" title=\"Bocarsly, M. E., Powell, E. S., Avena, N. M. &amp; Hoebel, B. G. High-fructose corn syrup causes characteristics of obesity in rats: increased body weight, body fat and triglyceride levels. Pharmacol. Biochem. Behav. 97, 101\u2013106 (2010).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref18\">18<\/a><\/sup>. Artificial honey produced exclusively from HFCS has a different carbohydrate profile (contains fructosyl-fructose) than artificial honey produced from sucrose<sup><a id=\"ref-link-23\" title=\"Ruiz-Matute, A. I., Weiss, M., Sammataro, D., Finely, J. &amp; Sanz, M. L. Carbohydrate Composition of High-Fructose Corn Syrups (HFCS) Used for Bee Feeding: Effect on Honey Composition. J. Agric. Food Chem. 58, 7317\u20137322 (2010).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref19\">19<\/a><\/sup>\u00a0and has been shown to decrease spring brood and wax production relative to sucrose<sup><a id=\"ref-link-24\" title=\"Sammataro, D. &amp; Weiss, M. Comparison of Productivity of Colonies of Honey Bees, Apis mellifera, Supplemented with Sucrose or High Fructose Corn Syrup. J. of Insect Sci. 13, 1\u201313 (2013).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref20\">20<\/a><\/sup>. Moreover, there is growing evidence that constituents in natural honey, absent from sucrose and HFCS, positively affect the honey bee\u2019s xenobiotic detoxification system<sup><a id=\"ref-link-25\" title=\"Johnson, R. M. et al. Ecologically appropriate xenobiotics induce cytochrome P450s in Apis mellifera. PLoS ONE 7, e31051\u2013 (2012).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref21\">21<\/a>,\u00a0<a id=\"ref-link-26\" title=\"Mao, W., Schuler, M. A. &amp; Berenbaum, M. R. Honey constituents up-regulate detoxification and immunity genes in the western honey bee Apis mellifera. Proc. Natl. Acad. Sci. USA 110, 8842\u20138846 (2013).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref22\">22<\/a><\/sup>. These results suggest that honey, sucrose and HFCS may impact honey bee physiology and health differently.<\/p>\n<p>We explored this issue further with whole-genome transcriptomics to comprehensively survey the effects of honey, sucrose and HFCS on fat body gene expression. The fat body is a multifunctional organ responsible for nutrient storage, energy mobilization and the production of antimicrobial peptides. Nutrient storage and mobilization are coupled to hormonal signals that include insulin and adipokinetic hormone to fulfill ongoing physiological demands<sup><a id=\"ref-link-27\" title=\"Arrese, E. L. &amp; Soulages, J. L. Insect fat body: energy, metabolism, and regulation. Annu. Rev. Entomol. 55, 207\u2013225 (2010).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref23\">23<\/a><\/sup>. In adult honey bees, the fat body is known to be transcriptionally responsive to nutritional manipulations and manipulations that affect aging and health<sup><a id=\"ref-link-28\" title=\"Ament, S. A. et al. Mechanisms of stable lipid loss in a social insect. J. Exp. Biol. 214, 3808\u20133821 (2011).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref24\">24<\/a>,\u00a0<a id=\"ref-link-29\" title=\"Alaux, C., Dantec, C., Parrinello, H. &amp; Le Conte, Y. Nutrigenomics in honey bees: digital gene expression analysis of pollen's nutritive effects on healthy and varroa-parasitized bees. BMC Genomics 12, 496 (2011).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref25\">25<\/a><\/sup>. We focused on the fat body to study the effects of different dietary carbohydrate sources on the expression of genes involved in hormonal signaling, nutrient storage, energy metabolism, and immune function. In this study, we used older bees (18\u201321 days old) because their diet is primarily carbohydrate-based<sup><a id=\"ref-link-30\" title=\"Crailsheim, K. et al. Pollen consumption and utilization in worker honeybees (Apis mellifera carnica): Dependence on individual age and function. J. Insect Physiol. 38, 409\u2013419 (1992).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref11\">11<\/a><\/sup>\u00a0and because older bees have been shown to be the primary consumers of carbohydrate supplements inside the hive<sup><a id=\"ref-link-31\" title=\"Brodschneider, R. &amp; Crailsheim, K. Nutrition and health in honey bees. Apidologie 41, 278\u2013294 (2010).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref5\">5<\/a><\/sup>.<\/p>\n<\/div>\n<\/div>\n<\/section>\n<section>\n<div id=\"results\">\n<h1><a>Results<\/a><\/h1>\n<div>\n<nav><\/nav>\n<p>Measurements taken daily throughout the week-long trials showed similar levels of food consumption for bees fed honey, HFCS or sucrose (0.040 \u00b1 0.001, 0.036 \u00b1 0.002, 0.032 \u00b1 0.003g\/bee\/day, respectively, F = 4.26\u00a0<i>P<\/i>\u00a0= 0.055). Mortality also did not vary between diets (F = 0.57\u00a0<i>P<\/i>\u00a0= 0.59) and was between 0\u20137% across all cages.<\/p>\n<p>RNA-sequencing (RNA-seq) was performed to examine the effect of each diet treatment on fat body gene expression. In total, 5 sample pools were sequenced per diet treatment and colony replicate (N = 10). Initial examination of our results revealed that one of our colony replicates was heavily infected with deformed wing virus (DWV). For Colony A, an average of 37.06 \u00b1 8.66% reads aligned to the DWV genome sequence compared to only 1.29 \u00b1 2.77% for Colony B. By contrast, an average of 53.81 \u00b1 0.08% of the RNA-seq reads mapped to the honey bee genome for Colony A, while for Colony B the average was 87.72 \u00b1 0.03%. Due to this difference, we analyzed each colony separately. We explored the effects of diet treatment using multidimensional scaling (MDS) plots (using the log fold change of the top 100 genes) generated for each colony. These analyses indicated that chronic feeding of sucrose and HFCS elicited transcriptomic profiles distinct from bees fed a honey diet (<a href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#supplementary-information\">Figure S1A and S1B<\/a>). This pattern was observed for both colonies, indicating similar transcriptomic responses to diet treatments occurred regardless of differences in apparent viral load.<\/p>\n<p>To probe more extensively for diet effects on gene expression, we analyzed the results from both colonies together, assessing the main effect of diet treatment with colony as a blocking factor. Honey elicited hundreds of differences in gene expression relative to HFCS and sucrose (<a href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#f1\">Figure 1A<\/a>). There were 104 genes differentially expressed (FDR &lt;0.1) in bees fed honey or HFCS and 220 genes differentially expressed between bees fed honey or sucrose. By contrast, differences between HFCS and sucrose diets were much more limited with a total of 8 genes differentially expressed between these two groups. Gene-wise comparisons show a substantial overlap (64 genes) between the honey vs. sucrose (29.1% overlapped) and honey vs. HFCS (61.5% genes overlapped) indicating strong similarities across these gene lists (<a href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#f1\">Figure 1B<\/a>).<\/p>\n<div id=\"f1\">\n<figure><figcaption>Figure 1: Differences in gene expression in honey bee fat body caused by diets of honey, sucrose or high-fructose corn syrup (HFCS).<\/figcaption><div>\n<p><a href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/fig_tab\/srep05726_F1.html\"><img decoding=\"async\" src=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/images_article\/srep05726-f1.jpg\" alt=\"Differences in gene expression in honey bee fat body caused by diets of honey, sucrose or high-fructose corn syrup (HFCS).\" \/><\/a><\/p>\n<div>\n<p>(A) Number of differentially expressed genes (DEGs) for each diet comparison (FDR&lt;0.1). (B) Number of DEGs that overlap among the diet comparisons. (C) Gene Ontology categories significantly enriched (<i>P<\/i>&lt;0.004) for each diet comparison.<\/p>\n<\/div>\n<ul>\n<li><a href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/fig_tab\/srep05726_F1.html\">Full size image (110 KB)<\/a><\/li>\n<\/ul>\n<\/div>\n<nav>\n<ul>\n<li><a href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/fig_tab\/srep05726_ft.html\">Figures\/tables index<\/a><\/li>\n<\/ul>\n<\/nav>\n<\/figure>\n<\/div>\n<p>Class prediction analyses using the support vector machine algorithm<sup><a id=\"ref-link-32\" title=\"Slawski, M., Daumer, M. &amp; Boulesteix, A.-L. CMA: a comprehensive Bioconductor package for supervised classification with high dimensional data. BMC Bioinformatics 9, 439 (2008).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref26\">26<\/a><\/sup>\u00a0revealed that diet-induced fat body gene expression changes were robust and consistent across samples. Class membership was predicted correctly with 96% (Honey vs. Sucrose), 97.5% (Honey vs. HFCS) and 100% (HFCS vs. Sucrose) accuracy, corresponding to sensitivity values (true positives identified) between 0.92\u20131 and specificity values (true negatives identified) of 0.95\u20131. Top predictors for each diet comparison are shown in\u00a0<a href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#supplementary-information\">Figure S2<\/a>. Notable among top predictor genes for the honey-based comparisons were\u00a0<i>glutathione S transferase O3<\/i>\u00a0(GB44803) and\u00a0<i>pale<\/i>\u00a0(GB40967), which are associated with xenobiotic detoxification and tyrosine metabolism, respectively<i>. Maltase B1<\/i>\u00a0(GB54549) and two other genes involved in energy metabolism, GB50596 (GO term: oxido-reductase activity) and GB48029 (GO term: acyl carnitine transporter activity), were top predictors for the HFCS vs. Sucrose comparison.<\/p>\n<p>Consistent with class prediction analyses, Gene Ontology (GO) enrichment analyses showed that the lists of genes upregulated by honey were enriched for genes involved in amino acid metabolism and oxidation reduction (<a href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#f1\">Figure 1C<\/a>,\u00a0<a href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#supplementary-information\">Table S1<\/a>), especially phenylalanine and tyrosine metabolism. These included\u00a0<i>pale, henna<\/i>\u00a0(GB48022) and\u00a0<i>homogentisate 1,2-dioxygenase<\/i>\u00a0(GB53288). Relative to sucrose, honey also upregulated the gene\u00a0<i>flavin monooxygenase<\/i>\u00a0<i>1<\/i>\u00a0(GB42239), which was associated with oxidation reduction and alkaloid detoxification<sup><a id=\"ref-link-33\" title=\"Naumann, C., Hartmann, T. &amp; Ober, D. Evolutionary recruitment of a flavin-dependent monooxygenase for the detoxification of host plant-acquired pyrrolizidine alkaloids in the alkaloid-defended arctiid moth Tyria jacobaeae. Proc. Natl. Acad. Sci. USA 99, 6085\u20136090 (2002).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref27\">27<\/a><\/sup>. By contrast, sucrose upregulated genes that were associated with axonogenesis, anion transport, and several transcription factors associated with organ development, such as\u00a0<i>doublesex<\/i>\u00a0(GB55036)<i>, knot<\/i>\u00a0(GB42304) and\u00a0<i>tolkin<\/i>(GB52106), while HFCS upregulated the transmembrane receptors,\u00a0<i>domeless<\/i>\u00a0(GB42244) and\u00a0<i>tyramine receptor<\/i>\u00a0(GB47385) (<a href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#f1\">Figure 1C<\/a>,\u00a0<a href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#supplementary-information\">Table S1<\/a>). No GO terms were enriched for the 8 genes differentially expressed between HFCS- and sucrose-fed bees.<\/p>\n<p>To gain further insights into the biological significance of these diet-induced differences in fat body gene expression, we compared our results to three previously published transcriptomic (microarray) experiments on nutritional aspects of behavioral maturation in honey bees<sup><a id=\"ref-link-34\" title=\"Ament, S. A. et al. Mechanisms of stable lipid loss in a social insect. J. Exp. Biol. 214, 3808\u20133821 (2011).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref24\">24<\/a><\/sup>. Behavioral maturation in honey bees involves a switch from a high protein to a high carbohydrate diet and a loss of approximately 50% of fat body lipids<sup><a id=\"ref-link-35\" title=\"Toth, A. L. &amp; Robinson, G. E. Worker nutrition and division of labour in honeybees. Anim. Behav. 69, 427\u2013435 (2005).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref13\">13<\/a><\/sup>\u00a0prior to the shift from working in the hive to foraging. This behavioral and physiological shift also includes a reduction in the blood titers of the lipid storage protein vitellogenin (Vg)<sup><a id=\"ref-link-36\" title=\"Nelson, C. M., Ihle, K. E., Fondrk, M. K., Page, R. E. &amp; Amdam, G. V. The gene vitellogenin has multiple coordinating effects on social organization. PLoS Biol. 5, e62 (2007).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref28\">28<\/a><\/sup>\u00a0which has been associated with the promotion of honey bee longevity<sup><a id=\"ref-link-37\" title=\"Seehuus, S.-C., Norberg, K., Gimsa, U., Krekling, T. &amp; Amdam, G. V. Reproductive protein protects functionally sterile honey bee workers from oxidative stress. Proc. Natl. Acad. Sci. USA 103, 962\u2013967 (2006).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref29\">29<\/a><\/sup>and immunity<sup><a id=\"ref-link-38\" title=\"Amdam, G. V. et al. Hormonal control of the yolk precursor vitellogenin regulates immune function and longevity in honeybees. Exp. Gerontol. 39, 767\u2013773 (2004).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref30\">30<\/a><\/sup>. We compared our results to three microarray experiments: 1) Maturation (hive bees compared to foragers); 2)\u00a0<i>vg<\/i>\u00a0knockdown (<i>vg<\/i>\u00a0RNAi compared to control); and 3) Diet [bees fed a high protein diet (45% pollen, 45% honey, 10% water) compared to sucrose (50% w\/v)]. We detected significant overlap between previously reported changes in fat body gene expression that occur during the hive to foraging transition<sup><a id=\"ref-link-39\" title=\"Ament, S. A. et al. Mechanisms of stable lipid loss in a social insect. J. Exp. Biol. 214, 3808\u20133821 (2011).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref24\">24<\/a><\/sup>\u00a0and our present Honey vs. Sucrose and Honey vs. HFCS gene lists (<a href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#t1\">Table 1<\/a>). Surprisingly, the fat body gene expression profile of the nutritionally enriched hive bees was more similar to that of the sucrose-fed bees, while the profile of the more nutritionally deprived foragers was more similar to that of the honey-fed bees (<a href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#t1\">Table 1<\/a>). GO enrichment analyses showed that the subset of genes overlapping the Honey vs. Sucrose or Honey vs. HFCS diets and the maturation-related experiment were associated with protein metabolism and oxidation reduction (<a href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#supplementary-information\">Table S2<\/a>). There also was a significant overlap between the Honey vs. Sucrose and Honey vs. HFCS gene lists and the gene list from the\u00a0<i>vg<\/i>\u00a0RNAi experiment<sup><a id=\"ref-link-40\" title=\"Ament, S. A. et al. Mechanisms of stable lipid loss in a social insect. J. Exp. Biol. 214, 3808\u20133821 (2011).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref24\">24<\/a><\/sup>; however, the log fold changes for overlapping genes were not significantly correlated indicating gene lists were not directionally similar (<a href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#t1\">Table 1<\/a>). There was no significant overlap between our Honey vs. Sucrose and Honey vs. HFCS gene lists and the Diet experiment<sup><a id=\"ref-link-41\" title=\"Ament, S. A. et al. Mechanisms of stable lipid loss in a social insect. J. Exp. Biol. 214, 3808\u20133821 (2011).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref24\">24<\/a><\/sup>, but the genes that did overlap showed a significant positive correlation that suggested honey\u2019s effects were directionally concordant to those of the Pollen + Honey treatment (<a href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#t1\">Table 1<\/a>).<\/p>\n<div id=\"t1\">\n<figure><figcaption>Table 1: Enrichment and correlation analyses for honey-related contrasts and previously published maturation-related microarray experiments. Pairwise comparisons are for genes differentially expressed at an FDR&lt;0.1. The number of genes compared is shown next to each experiment name in parentheses. RF is an enrichment factor for the number of genes that overlapped and\u00a0<i>P<\/i>\u00a0values indicate significant enrichment (hypergeometric test). Correlation results were calculated with genes that overlapped between pairwise comparisons<\/figcaption><div><a href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/fig_tab\/srep05726_T1.html\">Full table<\/a><\/div>\n<nav>\n<ul>\n<li><a href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/fig_tab\/srep05726_ft.html\">Figures\/tables index<\/a><\/li>\n<\/ul>\n<\/nav>\n<\/figure>\n<\/div>\n<\/div>\n<\/div>\n<\/section>\n<section>\n<div id=\"discussion\">\n<h1><a>Discussion<\/a><\/h1>\n<div>\n<p>A honey diet elicited a transcriptional profile distinct from sucrose and HFCS diets. These differences were present in two different honey bee colonies, with vastly different viral loads, indicating the impact of honey on fat body gene expression is robust. These results suggest that constituents in honey differentially regulate physiological processes and that sucrose and HFCS may not be equivalent nutritional substitutes to honey.<\/p>\n<p>Gene Ontology enrichment analyses showed honey upregulates genes associated with processes such as \u201caromatic amino acid family metabolic process,\u201d as well as \u201coxidation reduction.\u201d Among the genes in these categories were orthologs for the\u00a0<i>Drosophila melanogaster<\/i>\u00a0genes\u00a0<i>pale<\/i>\u00a0and\u00a0<i>henna<\/i>, which are related to phenylalanine and tyrosine metabolism. These amino acids have been linked to the production of neurotransmitters<sup><a id=\"ref-link-42\" title=\"Farooqui, T. Review of octopamine in insect nervous systems. Open Access Insect Physiol 4, 1\u201317 (2012).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref31\">31<\/a><\/sup>, and in the case of\u00a0<i>pale<\/i>\u00a0to immune responses to infection<sup><a id=\"ref-link-43\" title=\"De Gregorio, E., Spellman, P. T., Rubin, G. M. &amp; Lemaitre, B. Genome-wide analysis of the Drosophila immune response by using oligonucleotide microarrays. Proc. Natl. Acad. Sci. USA 98, 12590\u201312595 (2001).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref32\">32<\/a><\/sup>. Honey additionally upregulated the gene\u00a0<i>glutathione S transferase O3,<\/i>\u00a0whose activity is known to be induced by plant compounds and to have toxicological significance in the presence of pesticides<sup><a id=\"ref-link-44\" title=\"Yu, S. J. Insect glutathione S-transferases. Zool. Stud. 35, 9\u201319 (1996).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref33\">33<\/a><\/sup>. HFCS and sucrose relative to honey resulted in the upregulation of different biological processes. Sucrose, for example, upregulated processes such as axonogenesis but it is unlikely that axonogenesis is upregulated in our fat body samples; rather this GO category reflects upregulation of signaling pathways that play different roles in different tissues. HFCS upregulated the transmembrane receptors\u00a0<i>domeless<\/i>\u00a0and\u00a0<i>tyramine receptor<\/i>\u00a0suggesting differences in JAK-STAT signaling and tyrosine signaling between HFCS and honey.<\/p>\n<p>Sucrose and HFCS elicited a remarkably similar fat body transcriptional response. This result is consistent with previous studies showing no differences in colony productivity due to these diets<sup><a id=\"ref-link-45\" title=\"Severson, D. W. &amp; Erickson, E. H. Honey bee (Hymenoptera: Apidae) colony performance in relation to supplemental carbohydrates. J. of Econ. Entomol. 77, 1473\u20131478 (1984).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref15\">15<\/a><\/sup>but contrasts with findings showing differences in wax production and honey bee survival due to sucrose or HFCS<sup><a id=\"ref-link-46\" title=\"Barker, R. J. &amp; Lehner, Y. Laboratory comparison of high fructose corn syrup, grape syrup, honey and sucrose syrup as maintenance food for caged honey bees. Apidologie 9, 111\u2013116 (1978).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref14\">14<\/a>,\u00a0<a id=\"ref-link-47\" title=\"Sammataro, D. &amp; Weiss, M. Comparison of Productivity of Colonies of Honey Bees, Apis mellifera, Supplemented with Sucrose or High Fructose Corn Syrup. J. of Insect Sci. 13, 1\u201313 (2013).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref20\">20<\/a><\/sup>. Our results suggest that older bees may not be sensitive to increased fructose consumption because we detected little evidence that reflects the types of changes in hormonal signaling, energy metabolism and nutrient storage associated with high fructose corn syrup and increased fructose consumption in mammals<sup><a id=\"ref-link-48\" title=\"Bocarsly, M. E., Powell, E. S., Avena, N. M. &amp; Hoebel, B. G. High-fructose corn syrup causes characteristics of obesity in rats: increased body weight, body fat and triglyceride levels. Pharmacol. Biochem. Behav. 97, 101\u2013106 (2010).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref18\">18<\/a>,\u00a0<a id=\"ref-link-49\" title=\"Dekker, M. J., Su, Q. &amp; Baker, C. Fructose: a highly lipogenic nutrient implicated in insulin resistance, hepatic steatosis, and the metabolic syndrome. Am. J. of Physiol. Endocrinol Metab 299, E685-94 (2010).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref34\">34<\/a><\/sup>. Future research should test whether greater differences in gene expression due to sucrose or HFCS feeding are observed in young or middle-aged bees, which perform brood rearing and comb building tasks respectively, or whether the few differences in gene expression observed in this experiment can account for differences in colony performance in more natural conditions.<\/p>\n<p>To understand whether the gene expression differences associated with a honey diet relate to maturation-related physiological changes, we compared our results with previously published fat body microarray studies<sup><a id=\"ref-link-50\" title=\"Ament, S. A. et al. Mechanisms of stable lipid loss in a social insect. J. Exp. Biol. 214, 3808\u20133821 (2011).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref24\">24<\/a><\/sup>. These comparisons showed significant enrichment between honey-based gene lists and genes differentially expressed between hive bees vs. foragers, as well as with gene expression differences associated with\u00a0<i>vitellogenin<\/i>\u00a0RNAi treatment. Functional analyses suggest that shared changes in gene expression were related to protein metabolism and oxidation reduction, suggesting these processes are responsive to direct diet manipulations and maturational changes. Contrary to our expectations, we found the transcriptional profile of honey-fed-bees resembled the less well-nourished foragers rather than the more well-nourished nurse bees. These results suggest that there may be compounds in honey that modulate honey bee physiology towards a forager-like state.<\/p>\n<p>We did not find significant overlap between our Honey vs. Sugar differentially expressed gene list and the previously published list from bees that received either a diet of Pollen + Honey or sucrose. This result is surprising because the former contrast was embedded within this diet-related microarray experiment. This indicates that pollen is largely responsible for the gene expression changes in the diet microarray experiment and that those changes are separate from those elicited by honey in our experiment. Lack of statistical enrichment with pollen-induced changes partially reflects the relatively low level of protein in honey. In addition to the use of different experimental platforms (RNA-seq vs. microarray), there were also differences in the age of the bees assayed in each experiment: the diet-related microarray experiment investigated the effect of diet treatment on younger bees able to digest pollen, while our experiment assayed older bees with a decreased capability to digest pollen. Thus, the transcriptional differences elicited by a honey diet cannot be directly attributed to pollen traces in honey.<\/p>\n<p>Our goal was to perform a broad unbiased survey for the effects of honey, sucrose and HFCS on honey bee physiology. Our result that honey \u2013 but not sucrose or HFCS \u2013 upregulates genes associated with protein metabolism and oxidation reduction is indicative that honey elicits health-related physiological differences. We performed our experiment using older bees that typically consume sugar solutions inside the hive and do not digest pollen; therefore, constituents in honey may provide critical nutritional components and inducers that are otherwise absent in this age group. Previous research has already identified honey constituents that upregulate detoxification pathways in the gut<sup><a id=\"ref-link-51\" title=\"Mao, W., Schuler, M. A. &amp; Berenbaum, M. R. Honey constituents up-regulate detoxification and immunity genes in the western honey bee Apis mellifera. Proc. Natl. Acad. Sci. USA 110, 8842\u20138846 (2013).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref22\">22<\/a><\/sup>; our results further show honey induces gene expression changes on a more global scale. These changes may have toxicological relevance under natural conditions in contemporary agroecosystems, where bees are routinely exposed to toxins and pesticides.<\/p>\n<\/div>\n<\/div>\n<\/section>\n<section>\n<div id=\"methods\">\n<h1><a>Methods<\/a><\/h1>\n<div>\n<h2>Bees<\/h2>\n<p>We used bees from honey bee colonies from the University of Illinois Bee Research Facility, Urbana, IL, maintained according to standard beekeeping practices. The bees were a mixture of European subspecies typical of this region. To minimize genetic variation within a replicate, we used adult worker bees from a colony derived from a queen inseminated by single male; due to haplodiploidy, these bees were related to each other by an average coefficient of relatedness of 0.75. The experiment was replicated in two independent trials, each time using bees from different, unrelated, colony.<\/p>\n<h2>Feeding Trials<\/h2>\n<p>We used adult bees between the ages of 18\u201321 days old. These are older bees that readily consume various carbohydrate sources in the hive<sup><a id=\"ref-link-52\" title=\"Brodschneider, R. &amp; Crailsheim, K. Nutrition and health in honey bees. Apidologie 41, 278\u2013294 (2010).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref5\">5<\/a><\/sup>. To obtain focal bees we removed honeycomb frames containing pupae, placed them in an incubator (34\u00b0C\/30% RH), marked newly emerged one-day-old bees with a spot of paint (Testor\u2019s Paint, Rockford, IL, USA) on the dorsal surface of the thorax and reintroduced the marked bees into their natal colony; this was repeated three consecutive days to obtain a base population of &gt;500 marked bees in the hive. Focal bees were collected when they were 18\u201321 days old, placed into Plexiglas cages (10 \u00d7 10 \u00d7 7\u2005cm; 15 bees per cage) and assigned a diet treatment in the laboratory. Diet treatments consisted of 50% (w\/v) honey, 50% high fructose corn syrup (HFCS 55) or 50% sucrose\u00a0<i>ad libitum<\/i>\u00a0for each cage replicate (N = 3 cage replicates per treatment). All cages were kept in a 29\u00b0C incubator. Consumption and mortality were monitored daily for 7 days. After this time period, bees were flash-frozen and stored at \u221280\u00b0C for analysis.<\/p>\n<h2>Dissections and RNA extractions<\/h2>\n<p>Dissections were performed by incubating each abdomen in chilled RNA-later ICE (Ambion) at \u221220\u00b0C for a minimum of 16\u2005h. The gut and ventral tissue were then removed and RNA extraction performed on the fat body and adhering dorsal cuticle (RNeasy kit, Qiagen, with a DNAse treatment).<\/p>\n<h2>cDNA library construction and RNA-sequencing<\/h2>\n<p>We constructed cDNA libraries using pooled total RNA from fat body tissue from 3 individual bees. Pooling was performed to minimize sample variability within treatment groups. Each cDNA library was prepared with 1.5\u2005\u03bcg of pooled total RNA and constructed using the NEXTflex\u2122 Directional RNA-seq kit (Bioo Scientific) with an added mRNA purification step using Dynabeads\u00ae Oligo(dt)<sup><a id=\"ref-link-53\" title=\"Alaux, C., Dantec, C., Parrinello, H. &amp; Le Conte, Y. Nutrigenomics in honey bees: digital gene expression analysis of pollen's nutritive effects on healthy and varroa-parasitized bees. BMC Genomics 12, 496 (2011).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref25\">25<\/a><\/sup>\u00a0(Invitrogen). Library concentrations were quantified using Qubit\u00ae fluorometric quantitation and by quantitative real-time PCR using Kapa Library Quant kits (KapaBiosystems). Average fragment size and overall quality was evaluated with the Agilent 2100 Bioanalyzer platform and an Agilent High Sensitivity DNA kit. For sequencing, all libraries were diluted to a 6\u2005nM concentration. In total, we sequenced 5 libraries per treatment per colony, for a total of 30 libraries. Ten libraries were sequenced per lane (2\u20134 libraries per diet treatment\/per lane) with an Illumina HiSeq2000 instrument and were sequenced as single-end, 100\u2005nt reads.<\/p>\n<h2>Bioinformatics<\/h2>\n<p>RNA sequencing generated an average of 19,200,920 reads per library. Reads from each library were aligned against the\u00a0<i>Apis mellifera<\/i>\u00a0genome, Assembly 4.5<sup><a id=\"ref-link-54\" title=\"Elsik, C. G. et al. Finding the missing honey bee genes: lessons learned from a genome upgrade. BMC Genomics 15, 86 (2014).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref35\">35<\/a><\/sup>\u00a0using Tophat<sup><a id=\"ref-link-55\" title=\"Trapnell, C., Pachter, L. &amp; Salzberg, S. L. TopHat: discovering splice junctions with RNA-Seq. Bioinformatics 25, 1105\u20131111 (2009).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref36\">36<\/a><\/sup>\u00a0and read counts were generated for genes using HTSeq v0.5pv2 (<a href=\"http:\/\/www-huber.embl.de\/users\/anders\/HTSeq\">http:\/\/www-huber.embl.de\/users\/anders\/HTSeq<\/a>) and the\u00a0<i>Apis mellifera<\/i>\u00a0Official Gene Set, version 3.2. Reads that did not map uniquely or that mapped to genomic locations outside genes were not included in analyses for differential expression. We identified differentially expressed genes (DEGs) using a generalized linear model in the EdgeR package, version 3.2.4<sup><a id=\"ref-link-56\" title=\"Robinson, M. D. edgeR: a Bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics 26, 139\u2013140 (2010).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref37\">37<\/a><\/sup>.<\/p>\n<p>Because the number of differentially expressed genes in this study was in the hundreds instead of the thousands, we performed an additional analysis to test the robustness of the results. We compared differences in fat body gene expression between two subsets of the entire group of individuals fed on honey (n = 5 per subset), permuting the samples allocated to each subset a total of 100 times. We found only a few genes [2.07 \u00b1 0.344 genes (FDR&lt;0.1)] to be differentially expressed in these comparisons, suggesting that the differences we report between diet treatments are reliable.<\/p>\n<h2>Identification of Deformed Wing Virus Infection<\/h2>\n<p>Reads that did not align to the\u00a0<i>Apis mellifera<\/i>\u00a0genome were queried using BLAST on the NCBI website. BLAST results identified unaligned reads from Colony A as deformed wing virus (DWV) sequences. We further validated DWV infection by aligning all reads from each sample to the DWV genome (<a href=\"http:\/\/ftp.ncbi.nlm.nih.gov\/genomes\/Viruses\/Deformed_wing_virus_uid14891\">ftp.ncbi.nlm.nih.gov\/genomes\/Viruses\/Deformed_wing_virus_uid14891<\/a>), using Bowtie 2.<\/p>\n<h2>Class Prediction Analyses<\/h2>\n<p>To identify the most robust and consistent changes in fat body gene expression caused by each diet, we performed class prediction analyses using the support vector machine algorithm with 5-fold cross-validation. Class prediction analyses were performed using CMA<sup><a id=\"ref-link-57\" title=\"Slawski, M., Daumer, M. &amp; Boulesteix, A.-L. CMA: a comprehensive Bioconductor package for supervised classification with high dimensional data. BMC Bioinformatics 9, 439 (2008).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref26\">26<\/a><\/sup>, with normalized read counts for genes with a\u00a0<i>P<\/i>\u00a0value &lt;0.05. Top predictors for each diet contrast were selected based on their ranking as a top classifier in at least 10 of the 50 iterations.<\/p>\n<h2>Gene Ontology Analysis<\/h2>\n<p>Inferences for major functional themes for each DEG list were drawn from GO enrichment analyses using the DAVID Bioinformatic Resources 6.7 functional annotation tool<sup><a id=\"ref-link-58\" title=\"Dennis, G. et al. DAVID: Database for Annotation, Visualization, and Integrated Discovery. Genome Biol. 4, P3 (2003).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref38\">38<\/a><\/sup>. These analyses were performed using the\u00a0<i>Drosophila melanogaster<\/i>\u00a0orthologs associated with each DEG list. Statistical analyses of enrichment were performed with a hypergeometric test, using the number of honey bee genes with annotated\u00a0<i>Drosophila<\/i>\u00a0orthologs as the reference.<\/p>\n<h2>Comparisons to Previous Microarray Studies<\/h2>\n<p>Ament et al. (2011) previously published the results of microarray experiments investigating fat body gene expression related to nutritional aspects of behavioral maturation. We compared our results to three of them: 1) Maturation (hive bees compared to foragers; 2)\u00a0<i>vitellogenin<\/i>\u00a0knockdown (<i>vitellogenin<\/i>\u00a0dsRNA compared to control; and 3) Diet: [bees fed a high-protein diet (45% pollen, 45% honey, 10% water) compared to sucrose (50% w\/v)].<\/p>\n<p>To determine whether the lists of DEGs contained significant levels of overlap, we calculated an enrichment factor (RF) by dividing the observed number of overlapping genes by the expected number. The expected number of overlapping genes was calculated by multiplying the length of each DEG list and then dividing this value by the total number of genes included in the analyses<sup><a id=\"ref-link-59\" title=\"Alaux, C. et al. Honey bee aggression supports a link between gene regulation and behavioral evolution. Proc. Natl. Acad. Sci. USA 106, 15400\u201315405 (2009).\" href=\"http:\/\/www.nature.com\/srep\/2014\/140717\/srep05726\/full\/srep05726.html#ref39\">39<\/a><\/sup>. An RF value greater than 1 indicates the observed number of overlapping genes is greater than the expected value, thus demonstrating enrichment. Significant enrichment was determined using a hypergeotmetric test (1-tailed) with the p-hyper function in R. To determine whether the overlapping genes between two lists were directionally concordant in the predicted direction, we compared the log-fold changes for each gene list and assessed significance with a Pearson\u2019s correlation. A positive and significant Pearson\u2019s correlation shows the majority of overlapping genes between two gene lists show concordant gene expression changes.<\/p>\n<\/div>\n<\/div>\n<\/section>\n<section>\n<div id=\"references\">\n<h1><a>References<\/a><\/h1>\n<div>\n<nav><\/nav>\n<ol>\n<li id=\"ref1\">Calderone, N. W. Insect Pollinated Crops, Insect Pollinators and US Agriculture: Trend Analysis of Aggregate Data for the Period 1992\u20132009. PLoS ONE 7, e37235 (2012).<\/li>\n<li id=\"ref2\">Committee on the Status of Pollinators in North America, National Research Council. [<i>Status of Pollinators in North America<\/i>]. (The National Academies Press, 2007).<\/li>\n<li id=\"ref3\">vanEngelsdorp, D., Hayes, J., Jr, Underwood, R. M. &amp; Pettis, J. A Survey of Honey Bee Colony Losses in the U.S., Fall 2007 to Spring 2008. 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Zool. Stud. 35, 9\u201319 (1996).<\/li>\n<li id=\"ref34\">Dekker, M. J., Su, Q. &amp; Baker, C. Fructose: a highly lipogenic nutrient implicated in insulin resistance, hepatic steatosis, and the metabolic syndrome. Am. J. of Physiol. Endocrinol Metab 299, E685-94 (2010).<\/li>\n<li id=\"ref35\">Elsik, C. G.\u00a0<i>et al.<\/i>\u00a0Finding the missing honey bee genes: lessons learned from a genome upgrade. BMC Genomics 15, 86 (2014).\n<ul>\n<li><a>t<\/a><\/li>\n<\/ul>\n<\/li>\n<li id=\"ref36\">Trapnell, C., Pachter, L. &amp; Salzberg, S. L. TopHat: discovering splice junctions with RNA-Seq. Bioinformatics 25, 1105\u20131111 (2009).<\/li>\n<li id=\"ref37\">Robinson, M. D. edgeR: a Bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics 26, 139\u2013140 (2010).<\/li>\n<li id=\"ref38\">Dennis, G.\u00a0<i>et al.<\/i>\u00a0DAVID: Database for Annotation, Visualization, and Integrated Discovery. Genome Biol. 4, P3 (2003).<\/li>\n<li id=\"ref39\">Alaux, C.\u00a0<i>et al.<\/i>\u00a0Honey bee aggression supports a link between gene regulation and behavioral evolution. Proc. Natl. Acad. Sci. USA 106, 15400\u201315405 (2009).<\/li>\n<\/ol>\n<\/div>\n<\/div>\n<\/section>\n<p>[\/vc_column_text][\/vc_column][\/vc_row]<\/p>\n","protected":false},"excerpt":{"rendered":"<p>[vc_row][vc_column][vc_column_text]Es un h\u00e1bito com\u00fan de la mayor\u00eda de los apicultores de sacar toda la miel de las colmenas y luego alimentarlas con jarabe de az\u00facar, con lo que pueden pasar&hellip;<\/p>\n","protected":false},"author":1,"featured_media":1339,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":[],"categories":[19,48,52,34],"tags":[37,41,64],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v20.10 - https:\/\/yoast.com\/wordpress\/plugins\/seo\/ -->\n<title>Jarabe de az\u00facar acorta la vida de las abejas - estudio cient\u00edfico - AbejasResistentes<\/title>\n<meta name=\"description\" content=\"Jarabe de az\u00facar acorta la vida de las abejas \u2013 estudio cient\u00edfico\" \/>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/espanol.resistantbees.es\/?p=131\" \/>\n<meta property=\"og:locale\" content=\"es_ES\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Jarabe de az\u00facar acorta la vida de las abejas - estudio cient\u00edfico - AbejasResistentes\" \/>\n<meta property=\"og:description\" content=\"Jarabe de az\u00facar acorta la vida de las abejas \u2013 estudio cient\u00edfico\" \/>\n<meta property=\"og:url\" content=\"https:\/\/espanol.resistantbees.es\/?p=131\" \/>\n<meta property=\"og:site_name\" content=\"AbejasResistentes\" \/>\n<meta property=\"article:publisher\" content=\"https:\/\/www.facebook.com\/profile.php?id=100006580005971\" \/>\n<meta property=\"article:published_time\" content=\"2019-06-28T19:56:36+00:00\" \/>\n<meta property=\"article:modified_time\" content=\"2025-06-25T19:34:31+00:00\" \/>\n<meta property=\"og:image\" content=\"http:\/\/espanol.resistantbees.es\/wp-content\/uploads\/sites\/2\/2019\/06\/OLBarrelFeeding.jpg\" \/>\n\t<meta property=\"og:image:width\" content=\"1024\" \/>\n\t<meta property=\"og:image:height\" content=\"681\" \/>\n\t<meta property=\"og:image:type\" content=\"image\/jpeg\" \/>\n<meta name=\"author\" content=\"beefree\" \/>\n<meta name=\"twitter:card\" content=\"summary_large_image\" \/>\n<meta name=\"twitter:creator\" content=\"@bioapi\" \/>\n<meta name=\"twitter:site\" content=\"@bioapi\" \/>\n<meta name=\"twitter:label1\" content=\"Escrito por\" \/>\n\t<meta name=\"twitter:data1\" content=\"beefree\" \/>\n\t<meta name=\"twitter:label2\" content=\"Tiempo de lectura\" \/>\n\t<meta name=\"twitter:data2\" content=\"24 minutos\" \/>\n<script type=\"application\/ld+json\" class=\"yoast-schema-graph\">{\"@context\":\"https:\/\/schema.org\",\"@graph\":[{\"@type\":\"Article\",\"@id\":\"https:\/\/espanol.resistantbees.es\/?p=131#article\",\"isPartOf\":{\"@id\":\"https:\/\/espanol.resistantbees.es\/?p=131\"},\"author\":{\"name\":\"beefree\",\"@id\":\"https:\/\/espanol.resistantbees.es\/#\/schema\/person\/3ebceda286102ef949fcf3ab84562502\"},\"headline\":\"Jarabe de az\u00facar acorta la vida de las abejas &#8211; 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