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Reciprocal selection pressures in host-parasite systems drive coevolutionary arms races that lead to advanced adaptations in both opponents. In the interactions between social parasites and their hosts, aggression is one of the major behavioural traits under selection. In a field manipulation, we aimed to disentangle the impact of slavemaking ants and nest density on aggression of Temnothorax longispinosus ants. An early slavemaker mating flight provided us with the unique opportunity to study the influence of host aggression and demography on founding decisions and success. We discovered that parasite queens avoided colony foundation in parasitized areas and were able to capture more brood from less aggressive host colonies. Host colony aggression remained consistent over the two months experiment, but did not respond to our manipulation. However, as a fifth of all host colonies were successfully invaded by parasite queens, slavemaker nest foundation acts as a strong selection event selecting for high aggression in host colonies., Organismic and Evolutionary Biology

There is growing concern that some managed and wild insect pollinator populations are in decline, potentially threatening biodiversity and sustainable food production on a global scale. In recent years, there has been increasing evidence that sub-lethal exposure to neurotoxic, neonicotinoid pesticides can negatively affect pollinator immunocompetence and could amplify the effects of diseases, likely contributing to pollinator declines. However, a direct pathway connecting neonicotinoids and immune functions remains elusive. In this study we show that haemocytes and non-neural tissues of the honeybee Apis mellifera express the building blocks of the nicotinic acetylcholine receptors that are the target of neonicotinoids. In addition, we demonstrate that the haemocytes, which form the cellular arm of the innate immune system, actively express choline acetyltransferase, a key enzyme necessary to synthesize acetylcholine. In a last step, we show that the expression of this key enzyme is affected by field-realistic doses of clothianidin, a widely used neonicotinoid. These results support a potential mechanistic framework to explain the effects of sub-lethal doses of neonicotinoids on the immune function of pollinators., Competing Interests: The authors declare that they have no competing interests. Tobias Pamminger is employed by Bayer AG., (© 2024 Pamminger et al.)

Environmental risk assessment traditionally relies on a wide range of in vivo testing to assess the potential hazards of chemicals in the environment. These tests are often time-consuming and costly and can cause test organisms' suffering. Recent developments of reliable low-cost alternatives, both in vivo- and in silico-based, opened the door to reconsider current toxicity assessment. However, many of these new approach methodologies (NAMs) rely on high-quality annotated genomes for surrogate species of regulatory risk assessment. Currently, a lack of genomic information slows the process of NAM development. Here, we present a phylogenetically resolved overview of missing genomic resources for surrogate species within a regulatory ecotoxicological risk assessment. We call for an organized and systematic effort within the (regulatory) ecotoxicological community to provide these missing genomic resources. Further, we discuss the potential of a standardized genomic surrogate species landscape to enable a robust and nonanimal-reliant ecotoxicological risk assessment in the systems ecotoxicology era.

Plant protection products to be placed on the market in the European Union need to meet rigorous safety criteria including the testing of lumbricid earthworms, the functionally most important soil organism group in Central European agricultural ecosystems. To address uncertainties and investigate the potential long-term in-crop effects of the fungicide Cantus ® containing 50% boscalid as an active substance, a series of standardized earthworm field studies with an overall duration of 5 years per study program was carried out in four German agricultural fields under realistic crop rotation conditions. A two-step approach was chosen to analyze the potential overall long-term effects on earthworms in agricultural fields: (i) an assessment of the earthworm abundance development in the course of the four study programs in relation to the determined actual content of boscalid in soil and (ii) an effect size meta-analysis of earthworm abundance 1 year after treatment for each consecutive year and study program. Measured boscalid concentrations in the soil after multiple applications were well above the maximum boscalid residues observed in agricultural soils across Central Europe. There were isolated statistically significant reductions of earthworm abundance for some species and groups at some time points during the studies, but no consistent relationship to the Cantus ® treatments was observed. These results were supported by the meta-analysis, indicating no adverse effects on earthworm populations. Therefore, fluctuations of abundance reflect the natural variation of the populations rather than a concentration-related response. Based on this comprehensive analysis, we conclude that there is no application rate-related effect of the 5-year use of Cantus ® on the development of the earthworm communities. The four study programs, paired with a comprehensive evaluation, directly address the concerns about the potential long-term effects of boscalid on earthworms in the field and suggest that multiyear applications do not adversely affect earthworm populations. Integr Environ Assess Manag 2022;18:1399-1413. © 2021 ECT Oekotoxikologie GmbH and BASF SE. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC)., (© 2021 ECT Oekotoxikologie GmbH and BASF SE. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).)

Intact soil food webs are pivotal to maintaining essential soil functions, such as carbon recycling, sequestering, and biomass production. Although the functional role of micro- (e.g., bacteria and fungi) and macrofauna (e.g., earthworms) is comparatively well established, the importance of the mesofauna community (e.g., abundance and diversity of Acari and Collembola) in maintaining soil functionality is less clear. We investigated this question in a six-month field experiment in arable soil by actively manipulating mesofauna abundance and biodiversity through the application of two legacy insecticides (lindane and methamidophos) at sufficiently high doses to reduce mesofauna abundance (well above previously registered application rates; 2.5 and 7.5 kg a.s./ha for lindane, and 0.6 and 3 kg a.s./ha for methamidophos) and measure the impact on organic matter degradation. Our results demonstrate that both insecticides had reduced Collembola and Acari abundances by up to 80% over the study's six-month duration. In addition, we observed less pronounced and more complex changes in mesofauna biodiversity over time. These included insecticide-dependent temporal fluctuations (both reduction and increase) for different estimates (indices) of local (alpha)-diversity over time and no lasting impact for most estimates after six months. Even at these exceptionally high field rates, Collembola and Acari diversity was observed to generally recover by six months. In contrast, considering organic matter breakdown, we found no evidence of a treatment-related effect. These results suggest that organic matter breakdown in arable soils is likely driven by other trophic levels (e.g., microorganisms or earthworms) with only a limited influence of the mesofauna community. We discuss these findings with regard to their implications for our current understanding of soil food web function and future European soil risk assessments. Integr Environ Assess Manag 2022;18:1423-1433. © 2021 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC)., (© 2021 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).)

Bees foraging in agricultural habitats can be exposed to plant protection products. To limit the risk of adverse events, a robust risk assessment is needed, which requires reliable estimates for the expected exposure. The exposure pathways to developing solitary bees in particular are not well described and, in the currently proposed form, rely on limited information. To build a scaling model predicting the amount of protein developing solitary bees need based on adult body weight, we used published data on the volume of pollen solitary bees provide for their offspring. This model was tested against and ultimately updated with additional literature data on bee weight and protein content of emerged bees. We rescaled this model, based on the known pollen protein content of bee-visited flowers, to predict the expected amount of pollen a generalist solitary bee would likely provide based on its adult body weight, and tested these predictions in the field. We found overall agreement between the models' predictions and the measured values in the field, but additional data are needed to confirm these initial results. Our study suggests that scaling models in the bee risk assessment could complement existing risk assessment approaches and facilitate the further development of accurate risk characterization for solitary bees; ultimately the models will help to protect them during their foraging activity in agricultural settings. Integr Environ Assess Manag 2022;18:308-313. © 2021 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC)., (© 2021 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).)

In many countries, the western honey bee is used as surrogate in pesticide risk assessments for bees. However, uncertainty remains in the estimation of pesticide risk to non-Apis bees because their potential routes of exposure to pesticides, life histories, and ecologies differ from those of honey bees. We applied the vulnerability concept in pesticide risk assessment to 10 bee species including the honey bee, 2 bumble bee species, and 7 solitary bee species with different nesting strategies. Trait-based vulnerability considers the evaluation of a species at the level of both the organism (exposure and effect) and the population (recovery), which goes beyond the sensitivity of individuals to a toxicant assessed in standard laboratory toxicity studies by including effects on populations in the field. Based on expert judgment, each trait was classified by its relationship to the vulnerability to pesticide exposure, effects (intrinsic sensitivity), and population recovery. The results suggested that the non-Apis bees included in our approach are potentially more vulnerable to pesticides than the honey bee due to traits governing exposure and population recovery potential. Our analysis highlights many uncertainties related to the interaction between bee ecology and the potential exposures and population-level effects of pesticides, emphasizing the need for more research to identify suitable surrogate species for higher tier bee risk assessments. Environ Toxicol Chem 2021;40:2640-2651. © 2021 SETAC., (© 2021 SETAC.)

Plant protection products, including insecticides, are important for global food production but can have adverse effects on nontarget organisms including bees. Historically, research investigating such effects has focused mainly on the honeybee (Apis mellifera), whereas less information is available for non-Apis bees. Consequently, a comprehensive hazard (sensitivity) assessment for the majority of bees is lacking, which in turn hinders accurate risk characterization and consequently bee protection. Interspecies sensitivity extrapolation based on body weight might be a way to improve the situation, but in the past such approaches often ignored the phylogenetic background of the species used, which in turn potentially reduces the robustness of such results. Published acute contact sensitivity data (median lethal dose per bee) of bees to insecticides, their body weight, and their phylogenetic background were used to build interspecies scaling models to predict bee sensitivity based on their weight. The results indicate that 1) bee body weight is a predictor of acute contact bee sensitivity to a range of insecticides, and 2) phylogeny (nonindependence of data points) needs to be considered in cross-species analysis, although it does not always confound the observed effects.  Environ Toxicol Chem 2021;40:2044-2052. © 2021 SETAC., (© 2021 SETAC.)

Bumblebees are constantly exposed to a wide range of biotic and abiotic stresses which they must defend themselves against to survive. Pathogens and pesticides represent important stressors that influence bumblebee health, both when acting alone or in combination. To better understand bumblebee health, we need to investigate how these factors interact, yet experimental studies to date generally focus on only one or two stressors. The aim of this study is to evaluate how combined effects of four important stressors (the gut parasite Nosema ceranae, the neonicotinoid insecticide thiamethoxam, the pyrethroid insecticide cypermethrin and the EBI fungicide tebuconazole) interact to affect bumblebees at the individual and colony levels. We established seven treatment groups of colonies that we pulse exposed to different combinations of these stressors for 2 weeks under laboratory conditions. Colonies were subsequently placed in the field for 7 weeks to evaluate the effect of treatments on the prevalence of N. ceranae in inoculated bumblebees, expression levels of immunity and detoxification-related genes, food collection, weight gain, worker and male numbers, and production of worker brood and reproductives. Exposure to pesticide mixtures reduced food collection by bumblebees. All immunity-related genes were upregulated in the bumblebees inoculated with N. ceranae when they had not been exposed to pesticide mixtures, and bumblebees exposed to the fungicide and the pyrethroid were less likely to have N. ceranae. Combined exposure to the three-pesticide mixture and N. ceranae reduced bumblebee colony growth, and all treatments had detrimental effects on brood production. The groups exposed to the neonicotinoid insecticide produced 40%-76% fewer queens than control colonies. Our findings show that exposure to combinations of stressors that bumblebees frequently come into contact with have detrimental effects on colony health and performance and could therefore have an impact at the population level. These results also have significant implications for current practices and policies for pesticide risk assessment and use as the combinations tested here are frequently applied simultaneously in the field. Understanding the interactions between different stressors will be crucial for improving our ability to manage bee populations and for ensuring pollination services into the future., (© 2020 British Ecological Society.)

Historically, bee regulatory risk assessment for pesticides has centred on the European honeybee (Apis mellifera), primarily due to its availability and adaptability to laboratory conditions. Recently, there have been efforts to develop a battery of laboratory toxicity tests for a range of non-Apis bee species to directly assess the risk to them. However, it is not clear whether the substantial investment associated with the development and implementation of such routine screening will actually improve the level of protection of non-Apis bees. We argue, using published acute toxicity data from a range of bee species and standard regulatory exposure scenarios, that current first-tier honeybee acute risk assessment schemes utilised by regulatory authorities are protective of other bee species and further tests should be conducted only in cases of concern. We propose similar analysis of alternative exposure scenarios (chronic and developmental) once reliable data for non-Apis bees are available to expand our approach to these scenarios. In addition, we propose that in silico (simulation) approaches can then be used to address population-level effects in more field-realistic scenarios. Such an approach could lead to a protective, but also workable, risk assessment for non-Apis species while contributing to pollination security in agricultural landscapes around the globe. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

To ease nutritional stress on managed as well as native bee populations in agricultural habitats, agro-environmental protection schemes aim to provide alternative nutritional resources for bee populations during times of need. However, such efforts have so far focused on quantity (supply of flowering plants) and timing (flower-scarce periods) while ignoring the quality of the two main bee relevant flower-derived resources (pollen and nectar). As a first step to address this issue we have compiled one geographically explicit dataset focusing on pollen crude protein concentration, one measurement traditionally associated with pollen quality for bees. We attempt to provide a robust baseline for protein levels bees can collect in- (crop and weed species) and off-field (wild plants) in agricultural habitats around the globe. Using this dataset we identify crops which provide sub-optimal pollen resources in terms of crude protein concentration for bees and suggest potential plant genera that could serve as alternative resources for protein. This information could be used by scientists, regulators, bee keepers, NGOs and farmers to compare the pollen quality currently offered in alternative foraging habitats and identify opportunities to improve them. In the long run, we hope that additional markers of pollen quality will be added to the database in order to get a more complete picture of flower resources offered to bees and foster a data-informed discussion about pollinator conservation in modern agricultural landscapes., Competing Interests: All authors are employed by BASF (an agricultural solution provider).

Parasites are an important selection pressure for all organisms, and host immune responses are key in shaping host-parasite interactions. Host species with strong immune defences may be expected to experience lower parasitism; on the other hand, investment in immune function is costly, so hosts that have evolved to invest more in immune defence may be expected to have been under greater selection pressure from parasites. Disentangling the coevolutionary dynamics requires comparative studies that quantify the immune responses of potential hosts of parasites in a community, but such studies are rare. Here, we studied the immune defences of six leaf-cutting ant species in a community for which their relationships with phorid fly parasitoid species are known. We tested whether the strength of the baseline immune defences of the different ant species correlated positively or negatively with parasitoid load (number and abundance of parasitoid species exploiting the ant species), and host specialization of parasitoid species (the proportion of specialist parasitoids using each host). We measured four immune variables: i) the encapsulation response to a standard challenge, levels of ii) phenoloxidase (PO) and iii) prophenoloxidae (PPO) immune enzymes, and iv) the number of haemocytes. We found that ant species differed in their encapsulation response, PO levels and number of haemocytes, and that there was a positive, not negative, correlation across ant species between the strength of several of the immune variables and parasitoid load, but not for host specialization. This is in keeping with the hypothesis that higher parasitoid load selects for greater investment in immune defences. Our results suggest that immunity may be an important factor accounting for the dynamics of host-parasitoid interactions in this community. Similar community-level studies may be insightful, both for understanding host-parasite community ecology and for applications such as biocontrol., (Copyright © 2019. Published by Elsevier Ltd.)

Unequal reproductive output among members of the same sex (reproductive skew) is a common phenomenon in a wide range of communally breeding animals. In such species, reproductive dominance is often acquired during antagonistic interactions between group members that establish a reproductive hierarchy in which only a few individuals reproduce. Rank-specific syndromes of behavioural and physiological traits characterize such hierarchies, but how antagonistic behavioural interactions translate into stable rank-specific syndromes remains poorly understood. The pleiotropic nature of hormones makes them prime candidates for generating such syndromes as they physiologically integrate environmental (social) information, and often affect reproduction and behaviour simultaneously. Juvenile hormone (JH) is one of several hormones that occupy such a central regulatory role in insects and has been suggested to regulate reproductive hierarchies in a wide range of social insects including ants. Here we use experimental manipulation to investigate the effect of JH levels on reproductive physiology and social dominance in high-ranked workers of the eusocial ant Dinoponera quadriceps , a species that has secondarily reverted to queenless, simple societies. We show that JH regulated reproductive physiology, with ants in which JH levels were experimentally elevated having more regressed ovaries. In contrast, we found no evidence of JH levels affecting dominance in social interactions. This could indicate that JH and ovary development are decoupled from dominance in this species, however only high-ranked workers were investigated. The results therefore confirm that the regulatory role of JH in reproductive physiology in this ant species is in keeping with its highly eusocial ancestors rather than its secondary reversion to simple societies, but more investigation is needed to disentangle the relationships between hormones, behaviour and hierarchies., Competing Interests: The authors declare there are no competing interests.

There is growing concern that some bee populations are in decline, potentially threatening pollination security in agricultural and non-agricultural landscapes. Among the numerous causes associated with this trend, nutritional stress resulting from a mismatch between bee nutritional needs and plant community provisioning has been suggested as one potential driver. To ease nutritional stress on bee populations in agricultural habitats, agri-environmental protection schemes aim to provide alternative nutritional resources for bee populations during times of need. However, such efforts have focused mainly on quantity (providing flowering plants) and timing (during flower-scarce periods), while largely ignoring the quality of the offered flower resources. In a first step to start addressing this information gap, we have used literature data to compile a comprehensive geographically explicit dataset on nectar quality (i.e., total sugar concentration), offered to bees both within fields (crop and weed species) as well as outside fields (wild species) around the globe. Social bees are particularly sensitive to nectar sugar concentrations, which directly impact calorie influx into the colony and consequently their fitness making it an important resource quality marker. We find that the total nectar sugar concentrations in general do not differ between the three plant communities studied. In contrast we find increased variability in nectar quality in the wild plant community compared to crop and weed community, which is likely explained by the increased phylogenetic diversity in this category of plants. In a second step we explore the influence of local habitat on nectar quality and its variability utilizing a detailed sunflower ( Helianthus annuus L.) data set and find that geography has a small, but significant influence on these parameters. In a third step we identify crop groups (genera), which provide sub-optimal nectar resources for bees and suggest high quality alternatives as potential nectar supplements. In the long term this data set could serve as a starting point to systematically collect more quality characteristics of plant provided resources to bees, which ultimately can be utilized by scientist, regulators, NGOs and farmers to improve the flower resources offered to bees. We hope that ultimately this data will help to ease nutritional stress for bee populations and foster a data informed discussion about pollinator conservation in modern agricultural landscapes., Competing Interests: All authors work for BASF SE (an agricultural solution provider) and declare that they have no competing interests.

Social insects have evolved enormous capacities to collectively build nests and defend their colonies against both predators and pathogens. The latter is achieved by a combination of individual immune responses and sophisticated collective behavioral and organizational disease defenses, that is, social immunity. We investigated how the presence or absence of these social defense lines affects individual-level immunity in ant queens after bacterial infection. To this end, we injected queens of the ant Linepithema humile with a mix of gram+ and gram- bacteria or a control solution, reared them either with workers or alone and analyzed their gene expression patterns at 2, 4, 8, and 12 hr post-injection, using RNA-seq. This allowed us to test for the effect of bacterial infection, social context, as well as the interaction between the two over the course of infection and raising of an immune response. We found that social isolation per se affected queen gene expression for metabolism genes, but not for immune genes. When infected, queens reared with and without workers up-regulated similar numbers of innate immune genes revealing activation of Toll and Imd signaling pathways and melanization. Interestingly, however, they mostly regulated different genes along the pathways and showed a different pattern of overall gene up-regulation or down-regulation. Hence, we can conclude that the absence of workers does not compromise the onset of an individual immune response by the queens, but that the social environment impacts the route of the individual innate immune responses.

The evolution of complex societies with obligate reproductive division of labor represents one of the major transitions in evolution. In such societies, functionally sterile individuals (workers) perform many of fitness-relevant behaviors including allomaternal ones, without getting any direct fitness benefits. The question of how such worker division of labor has evolved remains controversial. The reproductive groundplan hypothesis (RGPH) offers a powerful proximate explanation for this evolutionary leap. The RGPH argues that the conserved genetic and endocrinological networks regulating fitness-relevant behavior (e g. foraging and brood care) in their solitary ancestors have become decoupled from actual reproduction in the worker caste and now generate worker behavioral phenotypes. However, the empirical support for this hypothesis remains limited to a handful of species making its general validity uncertain. In this study, we combine data from the literature with targeted sampling of key species and apply phylogenetically controlled comparative analysis to investigate if the key prediction of the RGPH, namely an association between allomaternal behavior and an allomaternal physiological state holds in the largest and most species-rich clade of social insects, the ants. Our findings clearly support the RPGH as a general framework to understand the evolution of the worker caste and shed light on one of the major transition in evolutionary history., (© 2016 The Author(s). Evolution © 2016 The Society for the Study of Evolution.)

Environmental temperature and temperature variation can have strong effects on the outcome of host-parasite interactions. Whilst such effects have been reported for different host systems, long-term consequences of pre-infection temperatures on host susceptibility and immunity remain understudied. Here, we show that experiencing both a biologically relevant increase in temperature and temperature variation undermines future disease susceptibility of the invasive garden ant Lasius neglectus when challenged with a pathogen under a constant temperature regime. In light of the economic and ecological importance of many social insects, our results emphasise the necessity to take the hosts' temperature history into account when studying host-parasite interactions under both natural and laboratory conditions, especially in the face of global change.

The ubiquitous trade-off between survival and costly reproduction is one of the most fundamental constraints governing life-history evolution. In numerous animals, gonadotropic hormones antagonistically suppressing immunocompetence cause this trade-off. The queens of many social insects defy the reproduction-survival trade-off, achieving both an extraordinarily long life and high reproductive output, but how they achieve this is unknown. Here we show experimentally, by integrating quantification of gene expression, physiology and behaviour, that the long-lived queens of the ant Lasius niger have escaped the reproduction-immunocompetence trade-off by decoupling the effects of a key endocrine regulator of fertility and immunocompetence in solitary insects, juvenile hormone (JH). This modification of the regulatory architecture enables queens to sustain a high reproductive output without elevated JH titres and suppressed immunocompetence, providing an escape from the reproduction-immunocompetence trade-off that may contribute to the extraordinary lifespan of many social insect queens., (© 2016 The Author(s).)

Division of labor is a defining characteristic of social insects and fundamental to their ecological success. Many of the numerous tasks essential for the survival of the colony must be performed at a specific location. Consequently, spatial organization is an integral aspect of division of labor. The mechanisms organizing the spatial distribution of workers, separating inside and outside workers without central control, is an essential, but so far neglected aspect of division of labor. In this study, we investigate the behavioral mechanisms governing the spatial distribution of individual workers and its physiological underpinning in the ant Myrmica rubra. By investigating worker personalities we uncover position-associated behavioral syndromes. This context-independent and temporally stable set of correlated behaviors (positive association between movements and attraction towards light) could promote the basic separation between inside (brood tenders) and outside workers (foragers). These position-associated behavior syndromes are coupled with a high probability to perform tasks, located at the defined position, and a characteristic cuticular hydrocarbon profile. We discuss the potentially physiological causes for the observed behavioral syndromes and highlight how the study of animal personalities can provide new insights for the study of division of labor and self-organized processes in general.

A new species of the ant genus Temnothorax Forel, 1890 - Temnothorax pilagens sp. n. is described from eastern North America. T. pilagens sp. n. is an obligate slave-making ant with two known hosts: T. longispinosus (Roger, 1863) and T. ambiguus (Emery, 1895). A differential diagnosis against Temnothorax duloticus (Wesson, 1937), the other dulotic congener from the Nearctic, is presented and a biological characteristics of the new species is given.

Adjusting to low temperatures is important for animals living in cold environments. We studied the chill-coma recovery time in temperate ant workers (Temnothorax nylanderi) from colonies collected in autumn and spring in Germany. We experimentally acclimated these ant colonies to cold temperatures followed by warm temperatures. As expected, cold-acclimated workers recovered faster from freezing temperatures, but subsequent heat acclimation did not change the short recovery times observed after cold acclimation. Hence, either heat acclimation improves cold tolerance, possibly as a general response to stress, or at least it does not negate enhanced cold tolerance following cold acclimation. Colonies collected in spring showed similar cold tolerance levels to cold-acclimated colonies in the laboratory. Next, we compared the chill-coma recovery time of different worker castes and found that exterior workers recovered faster than interior workers. This difference may be related to their more frequent exposure to cold, higher activity level, or distinct physiology. Interior workers were also heavier and showed a higher gaster-to-head ratio and thorax ratio compared to exterior workers. An obvious difference between exterior and interior workers is activity level, but we found no link between activity and cold tolerance. This suggests that physiology rather than behavioral differences could cause the increased cold tolerance of exterior workers. Our study reveals the importance of acclimation for cold tolerance under natural and standardized conditions and demonstrates differences in cold tolerance and body dimensions in monomorphic behavioral castes of an ant.

Reproductive division of labour is a characteristic trait of social insects. The dominant reproductive individual, often the queen, uses chemical communication and/or behaviour to maintain her social status. Queens of many social insects communicate their fertility status via cuticle-bound substances. As these substances usually possess a low volatility, their range in queen-worker communication is potentially limited. Here, we investigate the range and impact of behavioural and chemical queen signals on workers of the ant Temnothorax longispinosus. We compared the behaviour and ovary development of workers subjected to three different treatments: workers with direct chemical and physical contact to the queen, those solely under the influence of volatile queen substances and those entirely separated from the queen. In addition to short-ranged queen signals preventing ovary development in workers, we discovered a novel secondary pathway influencing worker behaviour. Workers with no physical contact to the queen, but exposed to volatile substances, started to develop their ovaries, but did not change their behaviour compared to workers in direct contact to the queen. In contrast, workers in queen-separated groups showed both increased ovary development and aggressive dominance interactions. We conclude that T. longispinosus queens influence worker ovary development and behaviour via two independent signals, both ensuring social harmony within the colony.

Slave-making ants reduce the fitness of surrounding host colonies through regular raids, causing the loss of brood and frequently queen and worker death. Consequently, hosts developed defenses against slave raids such as specific recognition and aggression toward social parasites, and indeed, we show that host ants react more aggressively toward slavemakers than toward nonparasitic competitors. Permanent behavioral defenses can be costly, and if social parasite impact varies in time and space, inducible defenses, which are only expressed after slavemaker detection, can be adaptive. We demonstrate for the first time an induced defense against slave-making ants: Cues from the slavemaker Protomognathus americanus caused an unspecific but long-lasting behavioral response in Temnothorax host ants. A 5-min within-nest encounter with a dead slavemaker raised the aggression level in T. longispinosus host colonies. Contrarily, encounters with nonparasitic competitors did not elicit aggressive responses toward non-nestmates. Increased aggression can be adaptive if a slavemaker encounter reliably indicates a forthcoming attack and if aggression increases postraid survival. Host aggression was elevated over 3 days, showing the ability of host ants to remember parasite encounters. The response disappeared after 2 weeks, possibly because by then the benefits of increased aggression counterbalance potential costs associated with it.

Hereditary sensory and autonomic neuropathy type II (HSAN II) leads to severe mutilations because of impaired nociception and autonomic dysfunction. Here we show that loss-of-function mutations in FAM134B, encoding a newly identified cis-Golgi protein, cause HSAN II. Fam134b knockdown results in structural alterations of the cis-Golgi compartment and induces apoptosis in some primary dorsal root ganglion neurons. This implicates FAM134B as critical in long-term survival of nociceptive and autonomic ganglion neurons.

Inhibitor 1 (I-1) is a protein inhibitor of protein phosphatase 1 (PP1), the predominating Ser/Thr phosphatase in the heart. Non-phosphorylated I-1 is inactive, whereas I-1 phosphorylated by protein kinase A (PKA) at Thr35 is a potent PP1 inhibitor. The phosphatases that dephosphorylate I-1Thr35 and thus deactivate I-1 in the heart are not established. Here we overexpressed I-1 in neonatal rat cardiac myocytes with recombinant adenovirus and determined phosphorylation of I-1, and one of the major target proteins of PKA/PP1 in the heart, phospholamban (PLB), by Western blot with phospho-specific antibodies. Incubation with the calcineurin inhibitor cyclosporine A or okadaic acid, used at a concentration preferentially inhibiting phosphatase 2A (PP2A), increased significantly I-1Thr35 (approximately 2- to 6-fold) and PLB Ser16 phosphorylation (approximately 2-fold). The results indicate that calcineurin and PP2A act to maintain a low basal level of phosphorylated (active) I-1 in living cardiac myocytes. Calcineurin may constitute a cross-talk between calcium- and cAMP-dependent pathways.

Objective: The protein phosphatase inhibitor-1 (I-1) is a highly specific and potent inhibitor of type 1 phosphatases (PP1) that is active only in its protein kinase A (PKA)-phosphorylated form. I-1 ablation decreases, I-1 overexpression sensitizes beta-adrenergic signaling in the heart. It is controversial whether I-1 expression is altered in human heart failure (HF), likely because its detection in heart is difficult due to its low abundance., Methods and Results: I-1 was >500-fold enriched from left ventricular myocardium (LVM) from patients with terminal HF (n=16) and non-failing controls (NF, n=5) and quantified with an affinity-purified I-1 and a I-1 phosphospecific antiserum. In non-failing I-1 protein levels amounted to 126 fmol/mg protein. In failing hearts, I-1 protein levels were reduced by 58% and I-1 phosphorylation by 77% (P<0.001 vs. NF). I-1 phosphorylation correlated well with serine-16 phosphorylation of phospholamban (PLB) in the same hearts (P<0.001). In contrast, PLB, troponin I (TnI) and PP1 protein and TnI phosphorylation levels did not differ between HF and NF., Conclusions: The results suggest that the reduction in I-1 protein and phosphorylation in failing human hearts leads to increased phosphatase activity which in turn may result in reduced phosphorylation of cardiac proteins such as PLB.

The protein phosphatase inhibitor-1 (PPI-1) inhibits phosphatase type-1 (PP1) only when phosphorylated by protein kinase A and could play a pivotal role in the phosphorylation/dephosphorylation balance. Rat cardiac PPI-1 was cloned by reverse transcriptase-polymerase chain reaction, expressed in Eschericia coli, evaluated in phosphatase assays, and used to generate an antiserum. An adenovirus was constructed encoding PPI-1 and green fluorescent protein (GFP) under separate cytomegalovirus promotors (AdPPI-1/GFP). A GFP-only virus (AdGFP) served as control. Engineered heart tissue (EHT) from neonatal rat cardiomyocytes and adult rat cardiac myocytes (ARCMs) were used as model systems. PPI-1 expression was determined in human ventricular samples by Northern blots. Compared with AdGFP, AdPPI-1/GFP-infected neonatal rat cardiomyocytes displayed a 73% reduction in PP1 activity. EHTs infected with AdPPI-1/GFP exhibited a fivefold increase in isoprenaline sensitivity. AdPPI-1/GFP-infected ARCMs displayed enhanced cell shortening as well as enhanced phospholamban phosphorylation when stimulated with 1 nM isoprenaline. PPI-1 mRNA levels were reduced by 57+/-12% in failing hearts with dilated and ischemic cardiomyopathy (n=8 each) compared with nonfailing hearts (n=8). In summary, increased PPI-1 expression enhances myocyte sensitivity to isoprenaline, indicating that PPI-1 acts as an amplifier in beta-adrenergic signaling. Decreased PPI-1 in failing human hearts could participate in desensitization of the cAMP pathway.