267 research outputs found
Behavioral alteration in the honeybee due to parasite-induced energetic stress
2012 Spring.Includes bibliographical references.Parasites are dependent on their hosts for energy and honeybee foragers with their high metabolic demand due to flight are especially prone to an energetic stress when they are infected. The microsporidian gut parasite Nosema ceranae is relatively new to the honeybee, Apis mellifera and because it is less co-evolved with its new host the virulence from infection can be particularly high. Using a series of feeding and survival experiments, I found that bees infected with N. ceranae have a higher appetite and hunger level, and the survival of infected bees is compromised when they are fed with a limited amount of food. However, if fed ad libitum the survival of infected individuals is not different from that of uninfected bees, demonstrating that energetic stress is the primary cause of the shortened lifespan observed in infected bees. I then developed a high throughput colorimetric assay to analyze hemolymph sugar levels of individual bees to demonstrate that the parasite mediated energetic stress is expressed as lower trehalose levels in free-flying bees, which suggests that infected bees are not only likely to have a reduced flight capacity but they are also unable to compensate for their lower energetic state. One of the ways in which the changing energetic state of an individual is predicted to impact its behavior is its sensitivity to risk although this has never been convincingly demonstrated. According to the energy budget rule of Risk Sensitivity Theory, it is adaptive for an animal to be risk averse when it is on a positive energy budget and be risk prone when it is on a negative budget because the utility of a potential large reward is much higher in the latter case. By constructing an empirical utility curve and conducting choice tests using a Proboscis Extension Response assay in bees that have been variously manipulated with respect to their energy budgets, I comprehensively demonstrated that bees shift between risk averse to risk prone behavior in accordance with the energy budge rule. Even more importantly, I showed that this shift is contingent upon a change in the energy budget as bees maintained on constant high or low energy budgets were found to be risk indifferent. Given that Nosema infected bees have been seen to forage precociously and inclement weather, my results suggest that such risky foraging might be a consequence of the lower energetic state of infected foragers. As these previous results suggest that parasitism, by lowering their energetic state could have a significant influence on how infected bees forage, I decided to test if the energetic state of an individual can regulate its foraging independent of the colony level regulation of foraging. I uncoupled the energetic state of the individual from that of the colony by feeding individual bees with the non-metabolizable sugar sorbose, thereby creating hungry bees in a satiated colony. I found that these energy depleted bees initially compensate for their lower energetic state by being less active within the colony and taking fewer foraging trips, but not by feeding more within the colony. However, with further depletion in their energetic state, these bees increase their foraging frequency showing that foraging is still partly regulated at the individual level even in a eusocial animal such as the honeybee. My research therefore shows that the energetic stress from a parasite could be a general mechanism that leads to significant behavioral alterations in infected individuals. Since the energetic state of an animal is a fundamental driver of its behavior, such a mechanism underlying behavioral alterations could have a significant impact on the life history of the host and transmission dynamics of a disease. More specifically, these results also suggest that a parasitic infection leading to energy depleted bees going out to forage in a risky manner also provides a plausible mechanism that explains the recent observations of bees disappearing from their colonies
The role of Janus Kinase 3 in CD4+ T Cell Homeostasis and Function: A Dissertation
This dissertation addresses the role for Janus Kinase 3 (Jak3) in CD4+ T cell homeostasis and function. Jak3 is a protein tyrosine kinase whose activity is essential for signals mediated by the Îłc dependent cytokines IL-2, -4, -7, -9, -15, and -21. Previous data have demonstrated that peripheral CD4+ T cells from Jak3-deficient mice have a memory phenotype and are functionally impaired in both proliferative and IL-2 responses in vitro. Interestingly, Jak3/Îłc activity has been previously shown to play a role in the prevention of T cell anergy.
These studies were initiated to more precisely define the role for Jak3/Îłc cytokines in the prevention of T cell anergy and the maintenance of functional CD4+ T cell responses. We began to address this question by assessing global gene expression changes between wild type and Jak3-/- CD4+ T cells. These data indicate that Jak3-/- CD4+ T cells have an increase in gene expression levels of inhibitory surface receptors as well as immunosuppressive cytokines.
Further analyses confirmed that Jak3-deficient T cells express high levels of PD-1, secrete a Trl-type cytokine profile following direct ex vivo activation, and suppress the proliferation of wild type T cells in vitro. These characteristics indicate that CD4+ Jak3-/- T cells share properties with regulatory T cell subsets that have an important role in peripheral tolerance and the prevention of autoimmunity.
We next addressed whether these regulatory characteristics were T cell intrinsic or rather the result of expanding in a Jak3-deficient microenvironment characterized by a number of immune abnormalities and a disrupted splenic architecture. Jak3-/- CD4+ T cells proliferate in vivoin a lymphopenic environment and selectively acquire regulatory T cell characteristics in the absence of any additional activation signals. While the precise mechanism by which Jak3-deficient T cells acquire these characteristics remains unclear, our data indicate that one important component is a T cell-intrinsic requirement for Jak3 signaling.
These findings indicate several interesting aspects of T cell biology. First, these studies, demonstrate that the homeostatic proliferation of CD4+ T cells is not dependent on signaling via Îłc-dependent cytokine receptors. And, second, that the weak activation signals normally associated with homeostatic expansion are sufficient to drive Jak3-/- T cells into a non-conventional differentiation program. Previous data indicate that, for wild type T cells, signaling through both the TCR as well as Îłc-dependent cytokine receptors promote the homeostatic proliferation of T cells in lymphopenic hosts. Since Jak3-/- T cells are unable to receive these cytokine signals, their proliferation is likely to be wholly dependent on TCR signaling. As a consequence of this TCR signaling, Jak3-/- T cells proliferate, but in addition, are induced to up regulate PD-1 and to selectively activate the IL-10 locus while shutting off the production of IL-2. Since this fate does not occur for wild type T cells in a comparable environment, it is likely that the unique differentiation pathway taken by Jak3-/- T cells reflects the effects of TCR signaling in the absence of Îłc-dependent cytokine signaling.
Interestingly, wild type T cells undergoing homeostatic expansion in lymphopenic hosts show many common patterns of gene expression to freshly-purified unmanipulated Jak3-/- T cells. For instance, micro array analysis of gene expression in wild type CD4+ T cells after lymphopenia induced homeostatic expansion show a similar pattern of upregulation in surface markers (PD-1 and LAG-3), and cytokine signaling molecules (IL-10 and IFN-Îł cytokine, receptors, and inducible gene targets) to that of Jak3-/- CD4+ T cells immediately ex vivo. These data suggest that the process of homeostatic proliferation normally induces immune attenuation and peripheral tolerance mechanisms, but that full differentiation into a regulatory T cell phenotype is prevented by Îłc-dependent cytokine signals.
Taken together these data suggest that Jak3 plays an important role in tempering typical immune attenuation mechanisms employed to maintain T cell homeostasis and peripheral tolerance
LD50 values may be misleading predictors of neonicotinoid toxicity across different bee species
The importance of not only honey bees (Apis mellifera) but also other non-managed bee species and their pollination services has come to light with their recently reported declines. One contributing factor in these declines is thought to be sub-lethal exposure to neonicotinoid insecticides such as thiacloprid. However, current government regulatory agencies do not require the assessment of insecticide toxicity on bee species other than the honey bee, even though previous studies have demonstrated that sensitivity to insecticides is not likely to be generalizable from honey bees to non-managed bee species. Replicating standardized protocols and testing five different doses of thiacloprid on individual caged bees, we assessed the acute contact toxicity by calculating mortality and the lethal dose (LD50) value for three bee species with different life history traits: Apis mellifera, Bombus terrestris, and Osmia bicornis. We found that Apis mellifera and Osmia bicornis had significantly higher mortality in comparison to Bombus terrestris, but there was no dose-dependent response for any of the three bee species. Bee size and sex were also not useful predictors of thiacloprid toxicity. These results suggest that solely relying on LD50 values, especially when they do not produce a dose-dependent response, may be misleading when assessing insecticide toxicity risk for honey bees and other non-managed bee species
Hive minded: like neurons, honeybees collectively integrate inhibitory stop signals to efficiently regulate forager recruitment
Social insects, such as ants and honeybees, and neurons in the brain, display collective decision-making, whereby groups of individuals or neurons collectively determine which stimuli to respond to and how to respond, yet crucially, no individual is aware that a decision is taking place. Honey bees rely on collective decision making when allocating foragers to the most profitable food source, based on positive reinforcement of the waggle dance. Here we examined whether negative feedback, via the audible stop signal, is being used at a collective level to more efficiently allocate of foragers to profitable food sources. We recorded feeder visits, waggle dances, waggle dance pheromones, and stop signals for bees that were marked and trained to a high 2.5 M sucrose solution feeder that was then switched to a poor food quality of 0.75 M sucrose solution. There was a burst of stop signals directed towards waggle dancing bees mostly from untrained individuals (contra signaling) right after the feeder switch, which then quickly returned back to baseline levels. Following this burst, waggle dancing (recruitment) and waggle dance pheromones continually decreased, but surprisingly previously trained bees visited the poor quality feeder more frequently. We then adjusted a neuron firing rate model with a sigmoidal gain-curve in order to model the dynamics of foragers waggle dancing for one of two potential food sites. We found that the addition of a brief spike of stop signals was enough to reproduce the dynamics of the focal population shown in the experiment. Our results suggest that: (1) honey bees can regulate foraging recruitment on a collective level independent from foraging frequency that appears to be regulated at an individual level and the two are not necessarily correlated, and (2) neural networks and honeybee colonies utilize similar network properties when making a decision
Honey bee (Apis mellifera) exposomes and dysregulated metabolic pathways associated with Nosema ceranae infection.
Honeybee Colony Vibrational Measurements to Highlight the Brood Cycle
Insect pollination is of great importance to crop production worldwide and honey bees are amongst its chief facilitators. Because of the decline of managed colonies, the use of sensor technology is growing in popularity and it is of interest to develop new methods which can more accurately and less invasively assess honey bee colony status. Our approach is to use accelerometers to measure vibrations in order to provide information on colony activity and development. The accelerometers provide amplitude and frequency information which is recorded every three minutes and analysed for night time only. Vibrational data were validated by comparison to visual inspection data, particularly the brood development. We show a strong correlation between vibrational amplitude data and the brood cycle in the vicinity of the sensor. We have further explored the minimum data that is required, when frequency information is also included, to accurately predict the current point in the brood cycle. Such a technique should enable beekeepers to reduce the frequency with which visual inspections are required, reducing the stress this places on the colony and saving the beekeeper time
Nosema spp. infections cause no energetic stress in tolerant honeybees
Host-pathogen coevolution leads to reciprocal adaptations, allowing pathogens to increase
host exploitation or hosts to minimise costs of infection. As pathogen resistance is often
associated with considerable costs, tolerance may be an evolutionary alternative. Here, we
examined the effect of two closely related and highly host dependent intracellular gut
pathogens, Nosema apis and Nosema ceranae, on the energetic state in Nosema tolerant and
sensitive honeybees facing the infection. We quantified the three major haemolymph
carbohydrates fructose, glucose, and trehalose using high-performance liquid
chromatography (HPLC) as a measure for host energetic state. Trehalose levels in the
haemolymph were negatively associated with N. apis infection intensity and with N. ceranae
infection regardless of the infection intensity in sensitive honeybees. Nevertheless, there was
no such association in Nosema spp. infected tolerant honeybees. These findings suggest that
energy availability in tolerant honeybees was not compromised by the infection. This result
obtained at the individual level may also have implications at the colony level where workers
in spite of a Nosema infection can still perform as well as healthy bees, maintaining colony
efficiency and productivity.The Deutsche Forschungsgemeinschaft
DFG priority programme SPP 1399 âHost-parasite co-evolutionâ (grant number MO373/26-
2).http://link.springer.com/journal/4362017-06-30hb2016Zoology and Entomolog
Honey bee (Apis mellifera) exposomes and dysregulated metabolic pathways associated with Nosema ceranae infection
Honey bee (Apis mellifera) health has been severely impacted by multiple environmental stressors including parasitic infection, pesticide exposure, and poor nutrition. The decline in bee health is therefore a complex multifactorial problem which requires a holistic investigative approach. Within the exposome paradigm, the combined exposure to the environment, drugs, food, and individuals' internal biochemistry affects health in positive and negative ways. In the context of the exposome, honey bee hive infection with parasites such as Nosema ceranae is also a form of environmental exposure. In this study, we hypothesized that exposure to xenobiotic pesticides and other environmental chemicals increases susceptibility to N. ceranae infection upon incidental exposure to the parasite. We further queried whether these exposures could be linked to changes in conserved metabolic biological pathways. From 30 hives sampled across 10 sites, a total of 2,352 chemical features were found via gas chromatography-time of flight mass spectrometry (GC-TOF) in extracts of honey bees collected from each hive. Of these, 20 pesticides were identified and annotated, and found to be significantly associated with N. ceranae infection. We further determined that infected hives were linked to a greater number of xenobiotic exposures, and the relative concentration of the exposures were not linked to the presence of a N. ceranae infection. In the exposome profiles of the bees, we also found chemicals inherent to known biological metabolic pathways of Apis mellifera and identified 9 dysregulated pathways. These findings have led us to posit that for hives exposed to similar chemicals, those that incur multiple, simultaneous xenobiotic stressors have a greater incidence of infection with N. ceranae. Mechanistically, our results suggests the overwhelming nature of these exposures negatively affects the biological functioning of the bee, and could explain how the decline in bee populations is associated with pesticide exposures
Effects of burial on leaf litter quality, microbial conditioning and palatability to three shredder taxa
1. Heterotrophic microorganisms are crucial for mineralising leaf litter and rendering it more palatable to leaf-shredding invertebrates. A substantial part of leaf litter entering running waters may be buried in the streambed and thus be exposed to the constraining conditions prevailing in the hyporheic zone. The fate of this buried organic matter and particularly the role of microbial conditioning in this habitat remain largely unexplored.
2. The aim of this study was to determine how the location of leaf litter within the streambed (i.e. at the surface or buried), as well as the leaf litter burial history, may affect the leaf-associated aquatic hyphomycete communities and therefore leaf consumption by invertebrate detritivores. We tested the hypotheses that (i) burial of leaf litter would result in lower decomposition rates associated with changes in microbial assemblages compared with leaf litter at the surface and (ii) altered microbial conditioning of buried leaf litter would lead to decreased quality and palatability to their consumers, translating into lower growth rates of detritivores.
3. These hypotheses were tested experimentally in a second-order stream where leaf-associated microbial communities, as well as leaf litter decomposition rates, elemental composition and toughness, were compared across controlled treatments differing by their location within the streambed. We examined the effects of the diverse conditioning treatments on decaying leaf palatability to consumers through feeding trials on three shredder taxa including a freshwater amphipod, of which we also determined the growth rate.
4. Microbial leaf litter decomposition, fungal biomass and sporulation rates were reduced when leaf litter was buried in the hyporheic zone. While the total species richness of fungal assemblages was similar among treatments, the composition of fungal assemblages was affected by leaf litter burial in sediment.
5. Leaf litter burial markedly affected the food quality (especially P content) of leaf material, probably due to the changes in microbial conditioning. Leaf litter palatability to shredders was highest for leaves exposed at the sediment surface and tended to be negatively related to leaf litter toughness and CâP ratio. In addition, burial of leaf litter led to lower amphipod growth rates, which were positively correlated with leaf litter P content.
6. These results emphasise the importance of leaf colonisation by aquatic fungi in the hyporheic zone of headwater streams, where fungal conditioning of leaf litter appears particularly critical for nutrient and energy transfer to higher trophic levels
Parasite-insecticide interactions: a case study of Nosema ceranae and fipronil synergy on honeybee
In ecosystems, a variety of biological, chemical and physical stressors may act in combination to induce illness in populations of living organisms. While recent surveys reported that parasite-insecticide interactions can synergistically and negatively affect honeybee survival, the importance of sequence in exposure to stressors has hardly received any attention. In this work, Western honeybees (Apis mellifera) were sequentially or simultaneously infected by the microsporidian parasite Nosema ceranae and chronically exposed to a sublethal dose of the insecticide fipronil, respectively chosen as biological and chemical stressors. Interestingly, every combination tested led to a synergistic effect on honeybee survival, with the most significant impacts when stressors were applied at the emergence of honeybees. Our study presents significant outcomes on beekeeping management but also points out the potential risks incurred by any living organism frequently exposed to both pathogens and insecticides in their habitat
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