Metabolic interactions control the transfer and spread of plasmid-encoded antibiotic resistance during surface-associated microbial growth.

Surface-associated microbial systems are hotspots for the spread of plasmid-encoded antibiotic resistance, but how surface association affects plasmid transfer and proliferation remains unclear. Surface association enables prolonged spatial proximities between different populations, which promotes plasmid transfer between them. However, surface association also fosters strong metabolic interactions between different populations, which can direct their spatial self-organization with consequences for plasmid transfer and proliferation. Here, we hypothesize that metabolic interactions direct the spatial self-organization of different populations and, in turn, regulate the spread of plasmid-encoded antibiotic resistance. We show that resource competition causes populations to spatially segregate, which represses plasmid transfer. In contrast, resource cross-feeding causes populations to spatially intermix, which promotes plasmid transfer. We further show that the spatial positionings that emerge from metabolic interactions determine the proliferation of plasmid recipients. Our results demonstrate that metabolic interactions are important regulators of both the transfer and proliferation of plasmid-encoded antibiotic resistance.

Bombus terrestris Prefer Mixed-Pollen Diets for a Better Colony Performance: A Laboratory Study.

Pollen is a major source of proteins and lipids for bumblebees. The nutritional content of pollen may differ from source plants, ultimately affecting colony development. This study investigated the foraging preferences of Bombus terrestris in regard to four pollen species, i.e., oilseed rape, wild apricot, sunflower, and buckwheat, under laboratory conditions. The results show that B. terrestris diversified their preference for pollens; the bumblebees mostly preferred wild apricot pollen, whereas sunflower pollen was the least preferred. The colonies fed on a mixed four-pollen diet, with a protein-lipid ratio of 4.55-4.86, exhibited better development in terms of the number of offspring, individual body size and colony weight. The colonies fed with buckwheat and sunflower pollens produced a significantly lower number of workers and failed to produce queen and male offspring. Moreover, wild apricot pollen had the richest protein content (23.9 g/100 g) of the four pollen species, whereas oilseed rape pollen had the highest lipid content (6.7 g/100 g), as revealed by the P:L ratios of wild apricot, sunflower, buckwheat, and oilseed rape, which were 6.76, 5.52, 3.50, and 3.37, respectively. Generally, B. terrestris showed feeding preferences regarding different pollens and a mixture of pollens, which ultimately resulted in differences in colony development. The findings of this study provide important baseline information to researchers and developers of nutritive pollen diets for bumblebees.

Colony performance of three native bumblebee species from South China and association with their gut microbiome.

Bumblebees play an important ecological economic role as pollinators in nature and agriculture. For reasons of biosecurity, many countries promote the cultivation of native bumblebee species for crop pollination instead of importing "alien" species. In South China, a few bumblebee species are considered useful in this way, particularly, Bombus atripes, Bombus bicoloratus and Bombus breviceps. However, whether they are suitable for artificial rearing and forming healthy colonies for pollination, remains unknown. In this project, queens from the 3 native species of Guizhou Province were collected and colonies were started under standardized conditions. The colonies were scored based on 19 parameters, including the stage of colony development, number and weight of offspring, and diet consumed. The data revealed that B. breviceps had the best performance, produced more workers and consumed the smallest diet. Next, we performed 16S rDNA sequencing of the bacterial communities found in the guts of offspring workers, and then a correlation analysis between colony performance and gut bacteria was conducted. Here, B. breviceps showed the highest diversity in gut bacterial composition, dominated by the bacteria Gilliamella, Snodgrassella, Enterobacter, and Lactobacillus Firm5. The higher the abundance of Snodgrassella, the better the performance of the colony in the foundation stage, and later Lactobacillus Firm5, Apibacter and Bifidobacterium were beneficial during the stages of rapid growth and colony decline. Although we do not understand all of the interactions yet, these correlations explain why B. breviceps demonstrated better colony performance. Our data provide valuable information for breeding local Bombus species and will contribute to developing strong colonies for crop pollination.

Evidence of Nest Reactivation and Perennial Colonies in the Neotropical Bumble Bee Bombus brevivillus (Hymenoptera: Apidae: Bombini).

Bombus brevivillus Franklin is a Neotropical Bombus species whose colonies are disappearing from most of its native range, and little is known about its biology and reproductive habits to help conservation or breeding efforts. Unlike Bombus species from temperate climates whose colonies perish every winter, there are suggestions of perennial colonies that Neotropical Bombus species can present. In this work, we investigated the development of two B. brevivillus colonies (i.e., number of workers, gynes, males, new cocoons, and brood area) between August 2012 and March 2013. We realized that while one colony collapsed and died after the males' and gynes' production, and the reduction of numbers of adult workers, in the other, a new queen assumed the posture at this phase and reactivated the old nest. Despite the reduced number of colonies investigated, this study shows the possibility of nest reactivation in the studied species in queen supersedure events in resemblance to perennial colonies of eusocial bees like Apis species and stingless bees. Such behavior has never been directly described to B. brevivillus in previous studies and opens the possibility for further research in the existence and the extension of perennial colonies in Neotropical Bombus species due to its importance to the species conservation in the tropical environment.

Exposure to elevated temperature during development affects bumblebee foraging behavior.

Bee foraging behavior provides a pollination service that has both ecological and economic benefits. However, bee population decline could directly affect the efficiency of this interaction. Among the drivers of this decline, global warming has been implicated as an emerging threat but exactly how increasing temperatures affect bee foraging behavior remains unexplored. Here, we assessed how exposure to elevated temperatures during development affects the foraging behavior and morphology of workers from commercial and wild Bombus terrestris colonies. Workers reared at 33 °C had a higher visiting rate and shorter visiting time than those reared at 27°C. In addition, far fewer workers reared at 33 °C engaged in foraging activities and this is potentially related to the drastic reduction in the number of individuals produced in colonies exposed to 33 °C. The impact of elevated developmental temperature on wild colonies was even stronger as none of the workers from these colonies performed any foraging trips. We also found that rearing temperature affected wing size and shape. Our results provide the first evidence that colony temperature can have striking effects on bumblebee foraging behavior. Of particular importance is the drastic reduction in the number of workers performing foraging trips, and the total number of foraging trips made by workers reared in high temperatures. Further studies should explore if, ultimately, these observed effects of exposure to elevated temperature during development lead to a reduction in pollination efficiency.

Lichen cell factories: methods for the isolation of photobiont and mycobiont partners for defined pure and co-cultivation.

BACKGROUND: Due to their huge biodiversity and the capability to produce a wide range of secondary metabolites, lichens have a great potential in biotechnological applications. They have, however, hardly been used as cell factories to date, as it is considered to be difficult and laborious to cultivate lichen partners in pure or co-culture in the laboratory. The various methods used to isolate lichen fungi, based on either the ascospores, the conidia, or the thallus, have so far not been compared or critically examined. Therefore, here we systematically investigate and compare the known methods and two new methods to identify the most suitable technology for isolation of fungi from lichens. RESULTS: Within this study six lichen fungi species were isolated and propagated as pure cultures. All of them formed colonies within one month. In case of lichens with ascocarps the spore discharge was the most suitable method. Spores were already discharged within 2 days and germinated within only four days and the contamination rate was low. Otherwise, the soredia and thallus method without homogenization, as described in this work, are also well suited to obtain pure fungal cultures. For the isolation of algae, we were also successful with the thallus method without homogenization. CONCLUSION: With the methods described here and the proposed strategic approach, we believe that a large proportion of the lichen fungi can be cultivated within a reasonable time and effort. Based on this, methods of controlled cultivation and co-cultivation must now be developed in order to use the potential of lichens with regard to their secondary metabolites, but also for other applications.

Five decades of misunderstanding in the social Hymenoptera: a review and meta-analysis of Michener's paradox.

In a much-cited 1964 paper entitled "Reproductive efficiency in relation to colony size in hymenopterous societies," Charles Michener investigated the correlation between a colony's size and its reproductive efficiency - the ability of its adult females to produce reproductives, measured as per-capita output. Based on his analysis of published data from destructively sampled colonies in 18 species, he reported that in most of these species efficiency decreased with increasing colony size. His conclusion that efficiency is higher in smaller groups has since gained widespread acceptance. But it created a seeming paradox: how can natural selection maintain social behaviour when a female apparently enjoys her highest per-capita output by working alone? Here we treat Michener's pattern as a hypothesis and perform the first large-scale test of its prediction across the eusocial Hymenoptera. Because data on actual output of reproductives were not available for most species, Michener used various proxies, such as nest size, numbers of brood, or amounts of stored food. We show that for each of Michener's data sets the reported decline in per-capita productivity can be explained by factors other than decreasing efficiency, calling into question his conclusion that declining efficiency is the cause of the pattern. The most prominent cause of bias is the failure of the proxy to capture all forms of output in which the colony invests during the course of its ontogeny. Other biasing factors include seasonal effects and a variety of methodological flaws in the data sets he used. We then summarize the results of 215 data sets drawn from post-1964 studies of 80 species in 33 genera that better control for these factors. Of these, 163 data sets are included in two meta-analyses that statistically synthesize the available data on the relationship between colony size and efficiency, accounting for variable sample sizes and non-independence among the data sets. The overall effect, and those for most taxonomic subgroups, indicates no loss of efficiency with increasing colony size. Two exceptional taxa, the halictid bees and independent-founding paper wasps, show negative trends consistent with the Michener hypothesis in some species. We conclude that in most species, particularly those with large colony sizes, the hypothesis of decreasing efficiency with increasing colony size is not supported. Finally, we explore potential mechanisms through which the level of efficiency can decrease, be maintained, or even increase, as colonies increase in size.

Early resources lead to persistent benefits for bumble bee colony dynamics.

Conditions experienced early in development can affect the future performance of individuals and populations. Demographic theories predict persistent population impacts of past resources, but few studies have experimentally tested such carry-over effects across generations or cohorts. We used bumble bees to test whether resource timing had persistent effects on within-colony dynamics over sequential cohorts of workers. We simulated a resource pulse for field colonies either early or late in their development and estimated colony growth rates during pulse- and non-pulse periods. During periods when resources were not supplemented, early-pulse colonies grew faster than late-pulse colonies; early-pulse colonies grew larger as a result. These results revealed persistent effects of past resources on current growth and support the importance of transient dynamics in natural ecological systems. Early-pulse colonies also produced more queen offspring, highlighting the critical nature of resource timing for the population, as well as colony, dynamics of a key pollinator.

Mitochondrial Retrograde Signaling Contributes to Metabolic Differentiation in Yeast Colonies.

During development of yeast colonies, various cell subpopulations form, which differ in their properties and specifically localize within the structure. Three branches of mitochondrial retrograde (RTG) signaling play a role in colony development and differentiation, each of them activating the production of specific markers in different cell types. Here, aiming to identify proteins and processes controlled by the RTG pathway, we analyzed proteomes of individual cell subpopulations from colonies of strains, mutated in genes of the RTG pathway. Resulting data, along with microscopic analyses revealed that the RTG pathway predominantly regulates processes in U cells, long-lived cells with unique properties, which are localized in upper colony regions. Rtg proteins therein activate processes leading to amino acid biosynthesis, including transport of metabolic intermediates between compartments, but also repress expression of mitochondrial ribosome components, thus possibly contributing to reduced mitochondrial translation in U cells. The results reveal the RTG pathway's role in activating metabolic processes, important in U cell adaptation to altered nutritional conditions. They also point to the important role of Rtg regulators in repressing mitochondrial activity in U cells.

Toxicity assessment of chronic exposure to common insecticides and bee medications on colony development and drones sperm parameters.

The effect of agrochemicals and beekeeping treatments on drones have not been widely investigated compared to workers or queens. In the present study, we investigated the chronic exposure of chemicals set (deltamethrin, acetamiprid, oxalic acid, fumagillin, and amitraz) on some sperm parameters and on the histomorphology of seminal vesicles. We also assessed the colony development and nosema load before and after the exposure. Thirty native Apis mellifera anatolica honeybee colonies with sister queens equalized with brood and total frame of bees were used for this experiment. Five colonies were used for each group. Deltamethrin, acetamiprid and fumagillin were given as oral chronic exposure at final concentrations of 25.10-6 mg L-1, 0.01 m L-1 and 50 mg L-1 respectively in syrup solution (50/50). Colonies were exposed to oxalic acid by spraying 5 mL per frame space of 3% (w/v) of oxalic acid dihydrate. Finally, the amitraz was applied based on the manufacturer's instructions. The concentrations chosen represented the field-realistic concentrations and those used by beekeepers in the region. Results showed that deltamethrin reduced brood production. In the same group, we found a high increase in nosema load. All treatments decreased sperm count except for fumagillin but this compound increased sperm mortality and increased the percentage of sperm with defected acrosome integrity. The amitraz exhibited a high sperm mortality and high percentage of sperm with defected membrane integrity function. The sperm parameters such as the count, the motility, the acrosome integrity, the membrane function of sperm, and the histomorphology of seminal vesicles of drones exposed to oxalic acid were the most affected. Bee medications commonly used such as oxalic acid and fumagillin should be more investigated and should be considered by beekeepers and particularly queen breeders.

The effect of DNA methylation on bumblebee colony development.

BACKGROUND: Although around 1% of cytosines in bees' genomes are known to be methylated, less is known about methylation's effect on bee behavior and fitness. Chemically altered DNA methylation levels have shown clear changes in the dominance and reproductive behavior of workers in queen-less colonies, but the global effect of DNA methylation on caste determination and colony development remains unclear, mainly because of difficulties in controlling for genetic differences among experimental subjects in the parental line. Here, we investigated the effect of the methylation altering agent decitabine on the developmental rate of full bumblebee colonies. Whole genome bisulfite sequencing was used to assess differences in methylation status. RESULTS: Our results showed fewer methylated loci in the control group. A total of 22 CpG loci were identified as significantly differentially methylated between treated and control workers with a change in methylation levels of 10% or more. Loci that were methylated differentially between groups participated in pathways including neuron function, oocyte regulation and metabolic processes. Treated colonies tended to develop faster, and therefore more workers were found at a given developmental stage. However, male production followed the opposite trend and it tended to be higher in control colonies. CONCLUSION: Overall, our results indicate that altered methylation patterns resulted in an improved cooperation between workers, while there were no signs of abnormal worker dominance or caste determination.

Evaluation of Antifungal Activity of Naja pallida and Naja mossambica Venoms against Three Candida Species.

In contrast to comprehensively investigated antibacterial activity of snake venoms, namely crude venoms and their selected components, little is known about antifungal properties of elapid snake venoms. In the present study, the proteome of two venoms of red spitting cobra Naja pallida (NPV) and Mozambique spitting cobra Naja mossambica (NMV) was characterized using LC-MS/MS approach, and the antifungal activity of crude venoms against three Candida species was established. A complex response to venom treatment was revealed. NPV and NMV, when used at relatively high concentrations, decreased cell viability of C. albicans and C. tropicalis, affected cell cycle of C. albicans, inhibited C. tropicalis-based biofilm formation and promoted oxidative stress in C. albicans, C. glabrata and C. tropicalis cells. NPV and NMV also modulated ammonia pulses during colony development and aging in three Candida species. All these observations provide evidence that NPV and NMV may diminish selected pathogenic features of Candida species. However, NPV and NMV also promoted the secretion of extracellular phospholipases that may facilitate Candida pathogenicity and limit their usefulness as anti-candidal agents. In conclusion, antifungal activity of snake venoms should be studied with great caution and a plethora of pathogenic biomarkers should be considered in the future experiments.

Characteristics of the Two Asian Bumblebee Species Bombus friseanus and Bombus breviceps (Hymenoptera: Apidae).

This study compared the food plants, life cycle, colony development, and mating behaviour of the two Asian bumblebee species Bombus friseanus and B. breviceps, which are very important pollinators for many wild flowers and crops in local ecosystems. Both species were shown to be highly polylectic. Differences were observed in their life cycles and colony development patterns. The colony foundation rate of the field-collected queens was high in both species, 95.5% in B. friseanus and 86.5% in B. breviceps. The intervals from colony initiation to colony sizes of 30, 60, and 80 workers and to the first male and gyne emergence were significantly shorter in B. friseanus than in B. breviceps (p < 0.01). The development period of the first batch of workers showed no significant difference between the two species (p > 0.05). Compared with B. friseanus, B. breviceps produced remarkably higher numbers of workers (135 ± 30 workers/colony in B. friseanus and 318 ± 123 workers/colony in B. breviceps) and males (199 ± 46 males/colony in B. friseanus and 355 ± 166 males/colony in B. breviceps) (p < 0.01), with notable variation was found among the colonies in both species. With no significant difference in the mating rate between these two species, the copulation duration of B. breviceps (1.54 ± 0.63 min) was strikingly shorter than that of B. friseanus (27.44 ± 11.16 min) (p < 0.001). This study highlights the characteristics of the two Asian bumblebee species and will aid further studies on their conservation and agricultural pollination use.

Effects of Climate Variability on Queen Production and Pollen Preferences of Neotropical Bumblebee Bombus atratus in a High Andean Suburban Condition.

Bombus atratus Franklin is a widely distributed bumblebee of South America. In Colombia, this species is recognized for its ability to adapt to highly disturbed habitats. However, knowledge of its ecology is poorly known, in particular conditions to ensure the long-term conservation of its populations. Identification of pollen resources is an important issue that could be used as a tool to manage and conserve bumblebees. In tropical areas, rainfall patterns could affect floral phenology and therefore the availability of pollen resources. Considering this, the present work aimed to establish the effect of extreme weather conditions (El Niño) in pollen availability, use of pollinic sources, and gyne production in B. atratus colonies. We reared and located 14 B. atratus colonies in a suburban area during a dry season (ENSO "El Niño") and a rainy season (ENSO "La Niña"). We registered time to gyne production and numbers of gynes produced per colony. We extracted pollen samples to establish both its floral origin and its relative abundance. We measured floral offer for each season. The data of pollen use per colony were utilized to perform Bipartite networks. We analyzed the production of gynes and pollen use per season with correlation models and generalized linear models. Colonies of the rainy season produced more gynes and faster. The floral diversity and offer were higher during the rainy season. Successful colonies used specific pollen sources in two seasons, independently of the floral offer. Extreme dry season affected development of B. atratus colonies.

Joint Toxicity of Acetamiprid and Co-Applied Pesticide Adjuvants on Honeybees under Semifield and Laboratory Conditions.

The evaluation of adverse effects of pesticides, pesticide adjuvants, and their combination on honeybees is hampered by a lack of colony-level bioassays reflecting productivity and survival over longer term exposure. In the present study, the joint toxicity of acetamiprid and co-applied pesticide adjuvants (N-methyl pyrrolidone [NMP], Silwet L-77, and Triton X-100) to honeybees was determined both in the laboratory and under semifield conditions. The 3 pesticide adjuvants caused no significant acute toxicity to honeybees by themselves; however, in the laboratory tests, they significantly increased the acute contact toxicity of acetamiprid to honeybees. For the semifield tests, in the T2 group (treatment with 5% acetamiprid soluble concentrate [SL] containing 10% Silwet L-77), the mortality of honeybees was significantly higher (p < 0.05) than that of the blank control on the fourth day after application (DAA + 4), that of the T1 group (5% acetamiprid SL containing 10% NMP) on DAA + 4 and DAA + 7 (seventh day after application), and that of the T3 group (5% acetamiprid SL containing 10% Triton X-100) on DAA + 4. Furthermore, the flight intensity in the T2 group on DAA + 7, the colony intensity on DAA + 28 (28th day after application), and the mean areas covered by pupae on DAA + 15 (15th day after application) were significantly lower (p < 0.05) than those of the blank control. Therefore, pesticide adjuvants may be important factors in increasing the toxicity of neonicotinoids to honeybees. Measures should be taken to manage the environmental risk of pesticide adjuvants during the process of formulation development and registration. Environ Toxicol Chem 2019;38:1940-1946. © 2019 SETAC.

Consistent pollen nutritional intake drives bumble bee (Bombus impatiens) colony growth and reproduction across different habitats.

Foraging behavior is a critical adaptation by insects to obtain appropriate nutrients from the environment for development and fitness. Bumble bees (Bombus spp.) form annual colonies which must rapidly increase their worker populations to support rearing reproductive individuals before the end of the season. Therefore, colony growth and reproduction should be dependent on the quality and quantity of pollen resources in the surrounding landscape. Our previous research found that B. impatiens foraging preferences to different plant species were shaped by pollen protein:lipid nutritional ratios (P:L), with foragers preferring pollen species with a ~5:1 P:L ratio. In this study, we placed B. impatiens colonies in three different habitats (forest, forest edge, and valley) to determine whether pollen nutritional quality collected by the colonies differed between areas that may differ in resource abundance and diversity. We found that habitat did not influence the collected pollen nutritional quality, with colonies in all three habitats collecting pollen averaging a 4:1 P:L ratio. Furthermore, there was no difference in the nutritional quality of the pollen collected by colonies that successfully reared reproductives and those that did not. We found however, that "nutritional intake," calculated as the colony-level intake rate of nutrient quantities (protein, lipid, and sugar), was strongly related to colony growth and reproductive output. Therefore, we conclude that B. impatiens colony performance is a function of the abundance of nutritionally appropriate floral resources in the surrounding landscape. Because we did not comprehensively evaluate the nutrition provided by the plant communities in each habitat, it remains to be determined how B. impatiens polylectic foraging strategies helps them select among the available pollen nutritional landscape in a variety of plant communities to obtain a balance of key macronutrients.

Imidacloprid application changes microbial dynamics and enzymes in rice soil.

Extensive use of imidacloprid in rice ecosystem may alter dynamics of microorganisms and can change soil biochemical properties. The objective of this study was to assess the effect of imidacloprid on growth and activities of microbes in tropical rice soil ecosystem. Four treatments, namely, recommended dose (at 25g a.i. ha-1, RD), double the recommended dose (at 50g a.i. ha-1, 2RD), five times the recommended dose (at 125g a.i. ha-1, 5RD) & ten times the recommended dose (at 250g a.i. ha-1, 10RD) along with control were imposed under controlled condition. Dissipation half lives of imidacloprid in soil were 19.25, 20.38, 21.65 and 33.00 days for RD, 2RD, 5RD and 10RD, respectively. In general bacteria, actinomycetes, fungi and phosphate solubilising bacteria population were disturbed due to imidacloprid application. Changes in diversity indices within bacterial community confirmed that imidacloprid application significantly affected distribution of bacteria. Total soil microbial biomass carbon content was reduced on imidacloprid application. Except dehydrogenase and alkaline phosphatase activities, all other soil enzymes namely, ß-glycosidase, fluorescien diacetate hydrolase, acid phosphatase and urease responded negatively to imidacloprid application. The extent of negative effect of imidacloprid depends on dose and exposure time. This study concludes imidacloprid application had transient negative effects on soil microbes.

Large-scale monitoring of effects of clothianidin-dressed oilseed rape seeds on pollinating insects in Northern Germany: effects on honey bees (Apis mellifera).

Possible effects of clothianidin seed-treated oilseed rape on honey bee colonies were investigated in a large-scale monitoring project in Northern Germany, where oilseed rape usually comprises 25-33 % of the arable land. For both reference and test sites, six study locations were selected and eight honey bee hives were placed at each location. At each site, three locations were directly adjacent to oilseed rape fields and three locations were situated 400 m away from the nearest oilseed rape field. Thus, 96 hives were exposed to fully flowering oilseed rape crops. Colony sizes and weights, the amount of honey harvested, and infection with parasites and diseases were monitored between April and September 2014. The percentage of oilseed rape pollen was determined in pollen and honey samples. After oilseed rape flowering, the hives were transferred to an extensive isolated area for post-exposure monitoring. Total numbers of adult bees and brood cells showed seasonal fluctuations, and there were no significant differences between the sites. The honey, which was extracted at the end of the exposure phase, contained 62.0-83.5 % oilseed rape pollen. Varroa destructor infestation was low during most of the course of the study but increased at the end of the study due to flumethrin resistance in the mite populations. In summary, honey bee colonies foraging in clothianidin seed-treated oilseed rape did not show any detrimental symptoms as compared to colonies foraging in clothianidin-free oilseed rape. Development of colony strength, brood success as well as honey yield and pathogen infection were not significantly affected by clothianidin seed-treatment during this study.

Comparison of buckwheat, red clover, and purple tansy as potential surrogate plants for use in semi-field pesticide risk assessments with Bombus impatiens.

Background. Bumble bees (Bombus spp.) are important wild and managed pollinators. There is increased interest in incorporating data on bumble bees into risk assessments for pesticides, but standardized methods for assessing hazards of pesticides in semi-field and field settings have not yet been established for bumble bees. During semi-field studies, colonies are caged with pesticide-treated flowering surrogate plants, which must be attractive to foragers to ensure colony exposure to the test compound, and must produce an ample nectar and pollen to sustain colonies during testing. However, it is not known which plant(s) are suitable for use in semi-field studies with bumble bees. Materials and Methods. We compared B. impatiens foraging activity and colony development on small plots of flowering buckwheat (Fagopyrum esculentum, var. common), red clover (Trifolium pratense), and purple tansy (Phacelia tanacetifolia) under semi-field conditions to assess their suitability as surrogate plants for pesticide risk assessment studies with bumble bees. We also compared the growth characteristics and input requirements of each plant type. Results. All three plant types generally established and grew well. Red clover and purple tansy experienced significant weed pressure and/or insect pest damage. In contrast, pest pressure was extremely low in buckwheat. Overall, B. impatiens foraging activity was significantly greater on buckwheat plots than red clover or purple tansy, but plant type had no effect on number of individuals produced per colony or colony weight. Discussion. Because of the consistently high foraging activity and successful colony development observed on buckwheat plots, combined with its favourable growth characteristics and low maintenance requirements, we recommend buckwheat as a surrogate plant for use in semi-field pesticide toxicity assessments with B. impatiens.

The Adenylate-Forming Enzymes AfeA and TmpB Are Involved in Aspergillus nidulans Self-Communication during Asexual Development.

Aspergillus nidulans asexual sporulation (conidiation) is triggered by different environmental signals and involves the differentiation of specialized structures called conidiophores. The elimination of genes flbA-E, fluG, and tmpA results in a fluffy phenotype characterized by delayed conidiophore development and decreased expression of the conidiation essential gene brlA. While flbA-E encode regulatory proteins, fluG and tmpA encode enzymes involved in the biosynthesis of independent signals needed for normal conidiation. Here we identify afeA and tmpB as new genes encoding members the adenylate-forming enzyme superfamily, whose inactivation cause different fluffy phenotypes and decreased conidiation and brlA expression. AfeA is most similar to unknown function coumarate ligase-like (4CL-Lk) enzymes and consistent with this, a K544N active site modification eliminates AfeA function. TmpB, identified previously as a larger homolog of the oxidoreductase TmpA, contains a NRPS-type adenylation domain. A high degree of synteny in the afeA-tmpA and tmpB regions in the Aspergilli suggests that these genes are part of conserved gene clusters. afeA, tmpA, and tmpB double and triple mutant analysis as well as afeA overexpression experiments indicate that TmpA and AfeA act in the same conidiation pathway, with TmpB acting in a different pathway. Fluorescent protein tagging shows that functional versions of AfeA are localized in lipid bodies and the plasma membrane, while TmpA and TmpB are localized at the plasma membrane. We propose that AfeA participates in the biosynthesis of an acylated compound, either a p-cuomaryl type or a fatty acid compound, which might be oxidized by TmpA and/or TmpB, while TmpB adenylation domain would be involved in the activation of a hydrophobic amino acid, which in turn would be oxidized by the TmpB oxidoreductase domain. Both, AfeA-TmpA and TmpB signals are involved in self-communication and reproduction in A. nidulans.