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      Colony Collapse Disorder (CCD) and bee age impact honey bee pathophysiology

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          Abstract

          Honey bee ( Apis mellifera) colonies continue to experience high annual losses that remain poorly explained. Numerous interacting factors have been linked to colony declines. Understanding the pathways linking pathophysiology with symptoms is an important step in understanding the mechanisms of disease. In this study we examined the specific pathologies associated with honey bees collected from colonies suffering from Colony Collapse Disorder (CCD) and compared these with bees collected from apparently healthy colonies. We identified a set of pathological physical characteristics that occurred at different rates in CCD diagnosed colonies prior to their collapse: rectum distension, Malpighian tubule iridescence, fecal matter consistency, rectal enteroliths (hard concretions), and venom sac color. The multiple differences in rectum symptomology in bees from CCD apiaries and colonies suggest effected bees had trouble regulating water. To ensure that pathologies we found associated with CCD were indeed pathologies and not due to normal changes in physical appearances that occur as an adult bee ages (CCD colonies are assumed to be composed mostly of young bees), we documented the changes in bees of different ages taken from healthy colonies. We found that young bees had much greater incidences of white nodules than older cohorts. Prevalent in newly-emerged bees, these white nodules or cellular encapsulations indicate an active immune response. Comparing the two sets of characteristics, we determined a subset of pathologies that reliably predict CCD status rather than bee age (fecal matter consistency, rectal distension size, rectal enteroliths and Malpighian tubule iridescence) and that may serve as biomarkers for colony health. In addition, these pathologies suggest that CCD bees are experiencing disrupted excretory physiology. Our identification of these symptoms is an important first step in understanding the physiological pathways that underlie CCD and factors impacting bee health.

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          Most cited references72

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          Diet effects on honeybee immunocompetence.

          The maintenance of the immune system can be costly, and a lack of dietary protein can increase the susceptibility of organisms to disease. However, few studies have investigated the relationship between protein nutrition and immunity in insects. Here, we tested in honeybees (Apis mellifera) whether dietary protein quantity (monofloral pollen) and diet diversity (polyfloral pollen) can shape baseline immunocompetence (IC) by measuring parameters of individual immunity (haemocyte concentration, fat body content and phenoloxidase activity) and glucose oxidase (GOX) activity, which enables bees to sterilize colony and brood food, as a parameter of social immunity. Protein feeding modified both individual and social IC but increases in dietary protein quantity did not enhance IC. However, diet diversity increased IC levels. In particular, polyfloral diets induced higher GOX activity compared with monofloral diets, including protein-richer diets. These results suggest a link between protein nutrition and immunity in honeybees and underscore the critical role of resource availability on pollinator health.
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            Pathogen Webs in Collapsing Honey Bee Colonies

            Recent losses in honey bee colonies are unusual in their severity, geographical distribution, and, in some cases, failure to present recognized characteristics of known disease. Domesticated honey bees face numerous pests and pathogens, tempting hypotheses that colony collapses arise from exposure to new or resurgent pathogens. Here we explore the incidence and abundance of currently known honey bee pathogens in colonies suffering from Colony Collapse Disorder (CCD), otherwise weak colonies, and strong colonies from across the United States. Although pathogen identities differed between the eastern and western United States, there was a greater incidence and abundance of pathogens in CCD colonies. Pathogen loads were highly covariant in CCD but not control hives, suggesting that CCD colonies rapidly become susceptible to a diverse set of pathogens, or that co-infections can act synergistically to produce the rapid depletion of workers that characterizes the disorder. We also tested workers from a CCD-free apiary to confirm that significant positive correlations among pathogen loads can develop at the level of individual bees and not merely as a secondary effect of CCD. This observation and other recent data highlight pathogen interactions as important components of bee disease. Finally, we used deep RNA sequencing to further characterize microbial diversity in CCD and non-CCD hives. We identified novel strains of the recently described Lake Sinai viruses (LSV) and found evidence of a shift in gut bacterial composition that may be a biomarker of CCD. The results are discussed with respect to host-parasite interactions and other environmental stressors of honey bees.
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              A Survey of Honey Bee Colony Losses in the U.S., Fall 2007 to Spring 2008

              Background Honey bees are an essential component of modern agriculture. A recently recognized ailment, Colony Collapse Disorder (CCD), devastates colonies, leaving hives with a complete lack of bees, dead or alive. Up to now, estimates of honey bee population decline have not included losses occurring during the wintering period, thus underestimating actual colony mortality. Our survey quantifies the extent of colony losses in the United States over the winter of 2007–2008. Methodology/Principal Findings Surveys were conducted to quantify and identify management factors (e.g. operation size, hive migration) that contribute to high colony losses in general and CCD symptoms in particular. Over 19% of the country's estimated 2.44 million colonies were surveyed. A total loss of 35.8% of colonies was recorded; an increase of 11.4% compared to last year. Operations that pollinated almonds lost, on average, the same number of colonies as those that did not. The 37.9% of operations that reported having at least some of their colonies die with a complete lack of bees had a total loss of 40.8% of colonies compared to the 17.1% loss reported by beekeepers without this symptom. Large operations were more likely to have this symptom suggesting that a contagious condition may be a causal factor. Sixty percent of all colonies that were reported dead in this survey died without dead bees, and thus possibly suffered from CCD. In PA, losses varied with region, indicating that ambient temperature over winter may be an important factor. Conclusions/Significance Of utmost importance to understanding the recent losses and CCD is keeping track of losses over time and on a large geographic scale. Given that our surveys are representative of the losses across all beekeeping operations, between 0.75 and 1.00 million honey bee colonies are estimated to have died in the United States over the winter of 2007–2008. This article is an extensive survey of U.S. beekeepers across the continent, serving as a reference for comparison with future losses as well as providing guidance to future hypothesis-driven research on the causes of colony mortality.

                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                17 July 2017
                2017
                : 12
                : 7
                : e0179535
                Affiliations
                [1 ] Department of Entomology, Plant Science Building University of Maryland, College Park, Maryland, United States of America
                [2 ] Cooperative Extension Butte County, University of California Cooperative Extension, Oroville, California, United States of America
                [3 ] Department of Ecology and Evolutionary Biology, The University of Arizona, Tucson, Arizona, United States of America
                [4 ] USDA-ARS, Bee Research Laboratory, Beltsville, Maryland, United States of America
                [5 ] Research Unit in Epidemiology and Risk Analysis applied to Veterinary Sciences (UREAR-ULg), Fundamental and Applied Research for Animal and Health (FARAH) Center, Faculty of Veterinary Medicine, University of Liège, Liège, Sart-Tilman, Belgium
                [6 ] USDA-ARS-PWA, Pollinating Insect-Biol., Mgmt. Syst.- Research Unit, Logan, Utah, United States of America and Department of Entomology, The Pennsylvania State University, University Park, Pennsylvania, United States of America
                Ghent University, BELGIUM
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                • Conceptualization: DVE DCF.

                • Data curation: DVE DCF MA EML KST.

                • Formal analysis: DVE DCF EML CS.

                • Funding acquisition: DVE DCF.

                • Investigation: DVE DCF MA.

                • Methodology: DVE DCF CS.

                • Project administration: DVE DCF.

                • Resources: DVE DCF MA YC.

                • Software: CS.

                • Supervision: DVE DCF.

                • Validation: YC KST.

                • Visualization: MA EML DCF KST.

                • Writing – original draft: MA DVE EML CS KST DCF.

                • Writing – review & editing: DVE KST MA EML YC CS DCF.

                Author information
                http://orcid.org/0000-0002-2418-1753
                Article
                PONE-D-16-43096
                10.1371/journal.pone.0179535
                5513415
                28715431
                7db06381-6528-4da5-9e1d-bd0c5cd98a4a

                This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

                History
                : 29 October 2016
                : 31 May 2017
                Page count
                Figures: 12, Tables: 3, Pages: 23
                Funding
                Funded by: Pennsylvania Department of Agriculture
                Award Recipient :
                Funded by: US National Honey Board
                Award Recipient :
                Funded by: The Pennsylvania State University
                Award Recipient :
                The following provided funding for this study: Pennsylvania Department of Agriculture (DVE, DCF), US National Honey Board (DVE, DCF), and the Pennsylvania State University, Department of Entomology (DCF, DVE). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Organisms
                Animals
                Invertebrates
                Arthropoda
                Insects
                Hymenoptera
                Bees
                Biology and Life Sciences
                Agriculture
                Agroecology
                Colony Collapse
                Biology and Life Sciences
                Ecology
                Agroecology
                Colony Collapse
                Ecology and Environmental Sciences
                Ecology
                Agroecology
                Colony Collapse
                Biology and Life Sciences
                Organisms
                Animals
                Invertebrates
                Arthropoda
                Insects
                Hymenoptera
                Bees
                Honey Bees
                Biology and Life Sciences
                Toxicology
                Toxic Agents
                Toxins
                Venoms
                Medicine and Health Sciences
                Pathology and Laboratory Medicine
                Toxicology
                Toxic Agents
                Toxins
                Venoms
                Engineering and Technology
                Management Engineering
                Decision Analysis
                Decision Trees
                Research and Analysis Methods
                Decision Analysis
                Decision Trees
                Biology and Life Sciences
                Plant Science
                Plant Anatomy
                Pollen
                Biology and Life Sciences
                Immunology
                Immune Response
                Medicine and Health Sciences
                Immunology
                Immune Response
                Biology and Life Sciences
                Anatomy
                Digestive System
                Gastrointestinal Tract
                Medicine and Health Sciences
                Anatomy
                Digestive System
                Gastrointestinal Tract
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                All relevant data are within the paper and its Supporting Information files.

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