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Abstract Anthropogenic pollution affects environments differently depending on proximity to pollution source, exposure route, and species ecology. Thus, understanding organism’s ecological role and exposure route to contaminants is central to assessing pollution impact. Treated municipal wastewater releases contaminants into waterways and alters microbial communities. Plants absorb contaminants and expose animals through foraging and nest-building activities. Nesting ecology differences of ground vs wood cavity-nesting bees offers insight into niche-specific susceptibility to pollution. Because contaminants bind to soil strongly, ground-nesting bees near wastewater are likely most impacted, while wood cavity-nesting bees likely less impacted since plants’ ability to uptake contaminants are species dependent. We compared gut microbiomes of directly exposed soil-nestingHalictus ligatusand indirectly exposed wood-nestingCeratinaspp. upstream/downstream of wastewater. We collected bees, flowers, and soil, and analyzed their bacteria microbiomes (16S rRNA). Wastewater altered ground-nestingH. ligatusmicrobiome >18 times greater than wood cavity-nestingCeratinaadults.Ceratinalarvae and pollen provisions showed significant but smaller shifts. Conversely, soil and flower microbiomes remained stable, indicating higher resilience. These results demonstrate that exposure routes drive contaminants susceptibility, with animal-associated microbes most vulnerable. Because bees are important pollinators and biodiversity contributors, these disruptions point to broader ecological risks in increasingly contaminated landscapes. Abstract Figure 

Theoretical frameworks of terrestrial community assembly often focus on single trophic levels (e.g. plants) without considering how complex interdependencies across different trophic levels influence assembly mechanisms. Yet, when multiple trophic levels are considered (e.g. plant–pollinator, plant–microbe interactions) the focus is typically on network analyses at local spatial scales. As spatial variation in biodiversity (β‐diversity) is increasingly being recognized for its relevance in understanding community assembly and conservation, considering how β‐diversity at one trophic level may be influenced by assembly processes that alter abundance and composition of interacting communities at a different trophic level (multitrophic dependency) is critical. Here, we build on single trophic level community assembly frameworks to explore the assembly processes affecting β‐diversity in multitrophic communities comprising flowering plants, their bee pollinators, and the corresponding bee‐gut microbiota to better understand the importance of multitrophic dependency in community assembly. Using distance‐based redundancy analysis and variation partitioning, we investigated community assembly processes across three interconnected trophic levels in two ecological regions in southern California: the Santa Monica Mountains and three islands of the Channel Island Archipelago. We found that the deterministic effects of multitrophic dependency are stronger on directly connected trophic levels than on indirectly connected trophic levels (i.e. flowers explain bee communities and bees explain bee‐gut bacteria communities, but flowers weakly explain variation in bee‐gut bacteria communities). We also found notable regional variation, where multitrophic dependency was weaker on the Channel Islands as ecological drift was more pronounced. Our results suggest that integrating the influence of multitrophic dependency on community assembly is important for elucidating drivers of β‐diversity and that multitrophic dependency can be determined by the regional context in which β‐diversity is measured. Taken together, our results highlight the importance of considering multiscale perspectives – both multitrophic and multiregional – in community assembly to fully elucidate assembly processes. 

ABSTRACT Social bees have been extensively studied for their gut microbial functions, but the significance of the gut microbiota in solitary bees remains less explored. Solitary bee,Megachile rotundatafemales provision their offspring with pollen from various plant species, harboring a diverse microbial community that colonizes larvae guts. TheApilactobacillusis the most abundant microbe, but evidence concerning the effects ofApilactobacillusand other provision microbes on growth and survival are lacking. We hypothesized that the presence ofApilactobacillusin abundance would enhance larval and prepupal development, weight, and survival, while the absence of intact microbial communities was expected to have a negative impact on bee fitness. We reared larvae on pollen provisions with naturally collected microbial communities (Natural pollen) or devoid of microbial communities (Sterile pollen). We also assessed the impact of introducingApilactobacillus micheneriby adding it to both types of pollen provisions. Feeding larvae with sterile pollen +A. micheneriled to the highest mortality rate, followed by natural pollen +A. micheneri, and sterile pollen. Larval development was significantly delayed in groups fed with sterile pollen. Interestingly, larval and prepupal weights did not significantly differ across treatments compared to natural pollen-fed larvae. 16S rRNA gene sequencing found a dominance ofSodalis, whenA. micheneriwas introduced to natural pollen. The presence ofSodaliswith abundantA. micheneri suggests potential crosstalk between both, shaping bee nutrition and health. Hence, this study highlights that the reliance on nonhost-specific environmental bacteria may not impact fitness ofM. rotundata.IMPORTANCEThis study investigates the impact of environmentally acquired gut microbes of solitary bee fitness with insights into the microbial ecology of bee and their health. While the symbiotic microbiome is well-studied in social bees, the role of environmental acquired microbiota in solitary bees remains unclear. Assessing this relationship in a solitary pollinator, the leaf-cutting bee,Megachile rotundata, we discovered that this bee species does not depend on the diverse environmental bacteria found in pollen for either its larval growth or survival. Surprisingly, high concentrations of the most abundant pollen bacteria, Apilactobacillus micheneridid not consistently benefit bee fitness, but caused larval mortality. Our findings also suggest an interaction betweenApilactobacillusand theSodalisand perhaps their role in bee nutrition. Hence, this study provides significant insights that contribute to understanding the fitness, conservation, and pollination ecology of other solitary bee species in the future. 

Pathogens and parasites of solitary bees have been studied for decades, but the microbiome as a whole is poorly understood for most taxa. Comparative analyses of microbiome features such as composition, abundance, and specificity, can shed light on bee ecology and the evolution of host–microbe interactions. Here we study microbiomes of ground-nesting cellophane bees (Colletidae: Diphaglossinae). From a microbial point of view, the diphaglossine genus Ptiloglossa is particularly remarkable: their larval provisions are liquid and smell consistently of fermentation. We sampled larval provisions and various life stages from wild nests of Ptiloglossa arizonensis and two species of closely related genera: Caupolicana yarrowi and Crawfordapis luctuosa . We also sampled nectar collected by P. arizonensis . Using 16S rRNA gene sequencing, we find that larval provisions of all three bee species are near-monocultures of lactobacilli. Nectar communities are more diverse, suggesting ecological filtering. Shotgun metagenomic and phylogenetic data indicate that Ptiloglossa culture multiple species and strains of Apilactobacillus , which circulate among bees and flowers. Larval lactobacilli disappear before pupation, and hence are likely not vertically transmitted, but rather reacquired from flowers as adults. Thus, brood cell microbiomes are qualitatively similar between diphaglossine bees and other solitary bees: lactobacilli-dominated, environmentally acquired, and non-species-specific. However, shotgun metagenomes provide evidence of a shift in bacterial abundance. As compared with several other bee species, Ptiloglossa have much higher ratios of bacterial to plant biomass in larval provisions, matching the unusually fermentative smell of their brood cells. Overall, Ptiloglossa illustrate a path by which hosts can evolve quantitatively novel symbioses: not by acquiring or domesticating novel symbionts, but by altering the microenvironment to favor growth of already widespread and generalist microbes. 

Despite the increasingly documented occurrence of individual specialization, the relationship between individual consumer interactions and diet-related microbial communities in wild populations is still unclear. Using data from nests of the bee Ceratina australensis from three different wild populations, we combine metabarcoding and network approaches to explore the existence of individual variation in resource use within and across populations, and whether dietary specialization affects the richness of pollen-associated microbes. We reveal the existence of marked dietary specialization. In the most specialized population, we also show that individuals' diet breadth was positively related to the richness of fungi, but not bacteria. Overall, individual specialization appeared to have a weak or negligible effect on the microbial richness of nests, suggesting that different mechanisms beyond environmental transmission may be at play regarding microbial acquisition in wild bees. 

Abstract Floral nectar, an important resource for pollinators, is inhabited by microbes such as yeasts and bacteria, which have been shown to influence pollinator preference. Dynamic and complex plant-pollinator-microbe interactions are likely to be affected by a rapidly changing climate, as each player has their own optimal growth temperatures and phenological responses to environmental triggers, such as temperature. To understand how warming due to climate change is influencing nectar microbial communities, we incubated a natural nectar microbial community at different temperatures and assessed the subsequent nectar chemistry and preference of the common eastern bumble bee, Bombus impatiens . The microbial community in floral nectar is often species-poor, and the cultured Brassica rapa nectar community was dominated by the bacterium Fructobacillus . Temperature increased the abundance of bacteria in the warmer treatment. Bumble bees preferred nectar inoculated with microbes, but only at the lower, ambient temperature. Warming therefore induced an increase in bacterial abundance which altered nectar sugars and led to significant differences in pollinator preference. 

ABSTRACT Diet and gut microbiomes are intricately linked on both short and long timescales. Changes in diet can alter the microbiome, while microbes in turn allow hosts to access novel diets. Bees are wasps that switched to a vegetarian lifestyle, and the vast majority of bees feed on pollen and nectar. Some stingless bee species, however, also collect carrion, and a few have fully reverted to a necrophagous lifestyle, relying on carrion for protein and forgoing flower visitation altogether. These “vulture” bees belong to the corbiculate apid clade, which is known for its ancient association with a small group of core microbiome phylotypes. Here, we investigate the vulture bee microbiome, along with closely related facultatively necrophagous and obligately pollinivorous species, to understand how these diets interact with microbiome structure. Via deep sequencing of the 16S rRNA gene and subsequent community analyses, we find that vulture bees have lost some core microbes, retained others, and entered into novel associations with acidophilic microbes found in the environment and on carrion. The abundance of acidophilic bacteria suggests that an acidic gut is important for vulture bee nutrition and health, as has been found in other carrion-feeding animals. Facultatively necrophagous bees have more variable microbiomes than strictly pollinivorous bees, suggesting that bee diet may interact with microbiomes on both short and long timescales. Further study of vulture bees promises to provide rich insights into the role of the microbiome in extreme diet switches. IMPORTANCE When asked where to find bees, people often picture fields of wildflowers. While true for almost all species, there is a group of specialized bees, also known as the vulture bees, that instead can be found slicing chunks of meat from carcasses in tropical rainforests. In this study, researchers compared the microbiomes of closely related bees that live in the same region but vary in their dietary lifestyles: some exclusively consume pollen and nectar, others exclusively depend on carrion for their protein, and some consume all of the above. Researchers found that vulture bees lost some ancestral “core” microbes, retained others, and entered into novel associations with acidophilic microbes, which have similarly been found in other carrion-feeding animals such as vultures, these bees’ namesake. This research expands our understanding of how diet interacts with microbiomes on both short and long timescales in one of the world’s biodiversity hot spots.