Metabolic Exchange Research Articles

Deciphering anammox response characteristics and potential mechanisms to polyethylene terephthalate microplastic exposure

Microplastics (MPs) are frequently detected in the wastewater. Herein, the short-term and long-term effects of polyethylene terephthalate (PET) MPs on anammox granular sludge were investigated and the potential response mechanisms were analyzed. Results showed that although short-term exposure of anammox granular sludge to PET-MPs induced a stress response, the nitrogen removal performance was not significantly affected. By contrast, long-term exposure to PET-MPs inhibited nitrogen removal performance with increased exposure time and PET-MP concentration. The total nitrogen removal efficiency (TNRE) decreased by 28.7% when sludge was exposed to 200mg/L of PET-MPs. However, the anammox activity recovered with prolonged operation time, and approximately 87% of the initial TNRE was recovered after three months. Microbial community evolution and metabolic exchange variations were the potential response mechanisms of anammox granular sludge to PET-MP exposure, with PET-MP exposure decreasing the anammox bacteria growth rate and relative symbiotic bacterial abundance in the anammox consortia and hindering cross-feeding pathways. The findings of this study provide novel insight into anammox behavior when treating wastewater containing PET-MPs.

Adaption mechanism and ecological role of CPR bacteria in brackish-saline groundwater

Candidate phyla radiation (CPR) constitutes a substantial fraction of bacterial diversity, yet their survival strategies and biogeochemical roles in brackish-saline groundwater remain unknown. By reconstructing 399 CPR metagenome-assembled genomes (MAGs) and 2007 non-CPR MAGs, we found that CPR, affiliated with 44 previously proposed phyla and 8 putative novel phyla, played crucial roles in maintaining the microbial stability and complexity in groundwater. Metabolic reconstructions revealed that CPR participated in diverse processes, including carbon, nitrogen, and sulfur cycles. Adaption of CPR to high-salinity conditions could be attributed to abundant genes associated with heat shock proteins, osmoprotectants, and sulfur reduction, as well as their cooperation with Co-CPR (non-CPR bacteria co-occurred with CPR) for metabolic support and resource exchange. Our study enhanced the understanding of CPR biodiversity in high-salinity groundwater, highlighting the collaborative roles of self-adaptive CPR bacteria and their reciprocal partners in coping with salinity stress, maintaining ecological stability, and mediating biogeochemical cycling.

Coral larvae increase nitrogen assimilation to stabilize algal symbiosis and combat bleaching under increased temperature.

Rising sea surface temperatures are increasingly causing breakdown in the nutritional relationship between corals and algal endosymbionts (Symbiodiniaceae), threatening the basis of coral reef ecosystems and highlighting the critical role of coral reproduction in reef maintenance. The effects of thermal stress on metabolic exchange (i.e., transfer of fixed carbon photosynthates from symbiont to host) during sensitive early life stages, however, remains understudied. We exposed symbiotic Montipora capitata coral larvae in Hawai'i to high temperature (+2.5°C for 3 days), assessed rates of photosynthesis and respiration, and used stable isotope tracing (4 mM 13C sodium bicarbonate; 4.5 h) to quantify metabolite exchange. While larvae did not show any signs of bleaching and did not experience declines in survival and settlement, metabolic depression was significant under high temperature, indicated by a 19% reduction in respiration rates, but with no change in photosynthesis. Larvae exposed to high temperature showed evidence for maintained translocation of a major photosynthate, glucose, from the symbiont, but there was reduced metabolism of glucose through central carbon metabolism (i.e., glycolysis). The larval host invested in nitrogen cycling by increasing ammonium assimilation, urea metabolism, and sequestration of nitrogen into dipeptides, a mechanism that may support the maintenance of glucose translocation under thermal stress. Host nitrogen assimilation via dipeptide synthesis appears to be used for nitrogen limitation to the Symbiodiniaceae, and we hypothesize that nitrogen limitation contributes to retention of fixed carbon by favoring photosynthate translocation to the host. Collectively, our findings indicate that although these larvae are susceptible to metabolic stress under high temperature, diverting energy to nitrogen assimilation to maintain symbiont population density, photosynthesis, and carbon translocation may allow larvae to avoid bleaching and highlights potential life stage specific metabolic responses to stress.

TMIC-52. DECODING METABOLIC INTERACTIONS IN GLIOBLASTOMA TUMOR MICROENVIRONMENT THROUGH COMPREHENSIVE SPATIAL TRANSCRIPTOMIC AND PROTEOMIC ANALYSIS

Abstract INTRODUCTION Within the GBM microenvironment, distinct micro-niches arise from independent cancer cell lineages with unique metabolic needs, characterized as Fast Cycling Cells (FCCs) and Slow Cycling Cells (SCCs). FCCs preferentially utilize aerobic glycolysis for energy, whereas treatment-resistant SCCs depend on lipid metabolism. Our study decodes the molecular and spatial heterogeneity within the GBM microenvironment, focusing on the dichotomy of immune infiltrates and the metabolic interactions between SCCs and the immune compartment in human GBM patients. METHODS We performed spatial transcriptomics and proteomics combined with geospatially resolved single-cell neighborhood analysis of IHC-stained GBM tissues using CellProfiler and the SNAQ algorithm that we developed to investigate the specific immune microenvironment of SCCs and FCCs. Holotomography and fluorescence-based time-lapse imaging coupled with flow cytometry were used to study lipid transfer between immune cells and GBM cells. Finally, the effects of pharmacological and genetic targeting of lipid transfer were investigated both in vitro and in vivo. RESULTS Our study establishes a connection between spatial metabolic heterogeneity and immune diversity, highlighting how specific tumor cell lineages correlate with characteristic immune micro-niches. In particular, SCCs exploit the metabolic properties of recruited immunosuppressive myeloid-derived cells, which metabolically support tumor cells by facilitating lipid transport and supporting SCC metabolism. Our results show that inhibiting lipid transfer affect SCCs, remodels the immune microenvironment, delays tumor growth and improves disease outcome. Our study also demonstrates a correlation between predicted sensitivity to lipid-modifying drugs, such as statins, and the SCC phenotype. This correlation could enable stratification and selection of patients most likely to benefit from such treatments. CONCLUSIONS This study establishes critical connections between the TME, cellular metabolism, anti-tumor immunity, and treatment vulnerabilities. Our research provides novel insights into how metabolic exchanges contribute to tumor progression and suggests that targeting metabolic interactions could be an effective therapeutic strategy for GBM.

Saccharomyces boulardii enhances anti-inflammatory effectors and AhR activation via metabolic interactions in probiotic communities.

Metabolic exchanges between strains in gut microbial communities shape their composition and interactions with the host. This study investigates the metabolic synergy between potential probiotic bacteria and Saccharomyces boulardii, aiming to enhance anti-inflammatory effects within a multi-species probiotic community. By screening a collection of 85 potential probiotic bacterial strains, we identified two strains that demonstrated a synergistic relationship with S. boulardii in pairwise co-cultivation. Furthermore, we computationally predicted cooperative communities with symbiotic relationships between S. boulardii and these bacteria. Experimental validation of 28 communities highlighted the role of S. boulardii as a key player in microbial communities, significantly boosting the community's cell number and production of anti-inflammatory effectors, thereby affirming its essential role in improving symbiotic dynamics. Based on our observation, one defined community significantly activated the aryl hydrocarbon receptor-a key regulator of immune response-280-fold more effectively than the community without S. boulardii. This study underscores the potential of microbial communities for the design of more effective probiotic formulations.

Disulfide-Rich Self-Assembling Peptides Based on Aromatic Amino Acid.

Aromatic residues in assembling peptides play a crucial role in driving peptide self-assembly through π-π stacking and hydrophobic interactions. Although various aromatic capping groups have been extensively studied, systematic investigations into the effects of single aromatic amino acids in assembling peptides remain limited. In this study, the influence of aromatic-aromatic interactions on disulfide-rich assembling peptides is systematically investigated by incorporating three different aromatic amino acids. Their folding propensity, self-assembling properties, and rheological behaviors are evaluated. These results indicate that different aromatic-aromatic interactions have a significant effect on self-assembly abilities, as determined by critical aggregation concentration (CAC) measurements. Furthermore, the biocompatibility of these hydrogels is assessed, and their potential for 3D cell culture is explored. The injectable F1-ox hydrogel demonstrate excellent biocompatibility for SHED and NIH3T3 cells and exhibit a porous structure that facilitates nutrient and cellular metabolic waste exchange. This work provides new insights into the molecular design of peptide-based biomaterials, with a focus on the impact of aromatic residues on disulfide-rich assembling peptides.

Integration of metagenomic analysis and metabolic modeling reveals microbial interactions in activated sludge systems in response to nanoplastics and plasticizers

Nanoplastics and plasticizers are prevalent in activated sludge and pose a potential threat to microbial communities in wastewater treatment systems. However, studies on the effects of nanoplastics and plasticizers on the interaction mechanisms and metabolic functions of microbial communities in activated sludge systems are still scarce. In this study, the responses of microbial interactions and metabolic functions to PVC nanoplastics (PVC-NPs) and bis(2-ethylhexyl) phthalate (DEHP) in activated sludge were investigated via a combination of amplicon sequencing, metagenome sequencing, and metabolic modeling. The results revealed that DEHP had a significant effect on the microbial community under short-term exposure. DEHP contamination may increase vitamin B12 producers to enhance species collaboration in communities. Furthermore, community metabolic modeling revealed that DEHP-degrading bacteria could promote positive interactions among community members. The increased metabolic exchange flux of siderophores and glutathione in microbial communities under PVC-NPs and DEHP contamination implied that microbial communities may maintain iron homeostasis in response to PVC-NPs and DEHP contamination through interspecies collaboration. However, more data are needed to further validate these results. This study provides vital insights into the response mechanisms of microbial interactions to nanoplastic and plasticizer contamination in activated sludge systems.

Effect of temperature on growth, metabolism, and gas exchange in Hermetia illucens larvae reared under commercial and laboratory conditions

Abstract The world’s socio-economic development is continuously increasing the demand for efficient production of food, feed, and energy from the agricultural sector. In this respect, the emerging production of black soldier fly larvae (BSFL) represents a promising system to upscale low-quality resources to higher-quality resources usable as feed or biodiesel. To optimize BSFL production it is critical to establish methods that are relevant to examine how rearing conditions, including temperature, affect growth, metabolism, and body composition at large scale. In a first set of experiments, we therefore compared how small-scale laboratory respirometry chambers (600 larvae) support gas exchange, growth, and metabolism in BSFL with similar measurements obtained from large-scale commercial conditions (>12 million larvae) at ∼34-35 °C. Having observed that small- and large-scale systems generated comparable measurements of growth and metabolism, we used our small-scale respirometry system to examine in detail how substrate rearing temperature ranging from 27.0 to 41.9 °C influenced growth, metabolism, and body composition in BSFL. Here we found that 7-day growth of BSFL was stable and high at 27, 35, and 39 °C, while a rearing temperature of ∼42 °C caused a severe, but sublethal, depression of growth and metabolism. Despite the general similarities in total growth at 27, 35, and 39 °C, we found considerable acceleration of metabolism and development at the two higher rearing temperatures (35 and 39 °C). In contrast, the lower rearing temperature (27 °C) resulted in reduced CO2 production per kg larvae produced and the was associated with a higher crude protein to crude lipid ratio in larvae after the 7-day growth period. Based on these findings we discuss how continued monitoring of gas exchange in production systems holds a potential to optimize production in the emerging industry of large-scale commercial insect production.

Stereolithography-assisted sodium alginate-collagen hydrogel scaffold with molded internal channels

Fabricating internal vascular networks within a hydrogel scaffold is essential for facilitating the supply of nutrients, oxygen, and metabolism exchange required by the encapsulated cells. The challenges in current hydrogel scaffold fabrication involve the difficulty of building adequate internal channels, poor scaffold geometry precision, and low cell viability caused by the fabrication process and polymer material properties. Stereolithography (SLA) stands out as a 3D printing technique distinguished by its superior production efficiency, advanced precision, and remarkable resolution in crafting intricate custom geometries. These attributes establish it as an innovative approach for templates in scaffold fabrication, potentially surpassing the fused deposition modeling (FDM)-based template strategy. Meanwhile, it exerts less shear stress on the cells compared to the direct bioprinting process. This novel strategy enables the fabrication of hydrogel vascular structure within the precision of 500 µm in both channel diameter and wall thickness. In this paper, various sodium alginate and collagen (SA-Col) composite hydrogels with varying collagen concentrations have been investigated to identify the optimal ratio for fabricating hydrogel scaffolds with channels.

Metabolic interdependencies in thermophilic communities are revealed using co-occurrence and complementarity networks

Microbial communities exhibit intricate interactions underpinned by metabolic dependencies. To elucidate these dependencies, we present a workflow utilizing random matrix theory on metagenome-assembled genomes to construct co-occurrence and metabolic complementarity networks. We apply this approach to a temperature gradient hot spring, unraveling the interplay between thermal stress and metabolic cooperation. Our analysis reveals an increase in the frequency of metabolic interactions with rising temperatures. Amino acids, coenzyme A derivatives, and carbohydrates emerge as key exchange metabolites, forming the foundation for syntrophic dependencies, in which commensalistic interactions take a greater proportion than mutualistic ones. These metabolic exchanges are most prevalent between phylogenetically distant species, especially archaea-bacteria collaborations, as a crucial adaptation to harsh environments. Furthermore, we identify a significant positive correlation between basal metabolite exchange and genome size disparity, potentially signifying a means for streamlined genomes to leverage cooperation with metabolically richer partners. This phenomenon is also confirmed by another composting system which has a similar wide range of temperature fluctuations. Our workflow provides a feasible way to decipher the metabolic complementarity mechanisms underlying microbial interactions, and our findings suggested environmental stress regulates the cooperative strategies of thermophiles, while these dependencies have been potentially hardwired into their genomes during co-evolutions.

3D Modeling: Insights into the Metabolic Reprogramming of Cholangiocarcinoma Cells.

Developing accurate in vitro models that replicate the in vivo tumor environment is essential for advancing cancer research and therapeutic development. Traditional 2D cell cultures often fail to capture the complex structural and functional heterogeneity of tumors, limiting the translational relevance of findings. In contrast, 3D culture systems, such as spheroids, provide a more physiologically relevant context by replicating key aspects of the tumor microenvironment. This study aimed to compare the metabolism of three intrahepatic cholangiocarcinoma cell lines in 2D and 3D cultures to identify metabolic shifts associated with spheroid formation. Cells were cultured in 2D on adhesion plates and in 3D using ultra-low attachment plates. Metabolic exchange rates were measured using NMR, and intracellular metabolites were analyzed using LC-MS. Significant metabolic differences were observed between 2D and 3D cultures, with notable changes in central carbon and glutathione metabolism in 3D spheroids. The results suggest that 3D cultures, which more closely mimic the in vivo environment, may offer a more accurate platform for cancer research and drug testing.

Effects of dietary nitrate, fumaric acid, and methanotrophic bacteria supplementation on rumino-intestinal nutrient metabolism and enteric gas exchange in dairy cows

The objective of the study was to investigate the effects of two approaches for enteric methane (CH4) mitigation and one approach for redirection of excess hydrogen (H2) resulting from CH4 inhibition, on dairy cows’ gas exchange and nutrient digestibility. Approaches for CH4 mitigation were methanotrophic bacteria supplementation and dietary nitrate, whereas the approach for H2 redirection was nitrate combined with fumaric acid. An incomplete 6 × 6 Latin square design experiment was conducted with 4 periods of 21 d using 6 rumen, duodenum, and ileum cannulated Danish Holstein cows with DIM of 123 ± 64.8 d (mean ± SD) and the milk yield was 33.6 ± 9.39 kg/d at beginning of experiment. The treatments were organized in a 2 × 3 factorial arrangement, where the first factor represented treatments without or with methanotrophic bacteria supplementation (MET), and the second factor represented 3 different dietary additive supplementations (DIET). These were a basal diet (BAS; no additives), a diet with nitrate (NIT; 10 g nitrate/kg dry matter (DM)), and a diet with nitrate combined with fumaric acid (NIT-F; NIT + 15 g fumaric acid/kg DM). Cows had ad libitum access to diets with a forage to concentrate ratio of 60 to 40 on dry matter basis. Following adaptation to experimental diets, samples of rumen fluid, digesta from duodenum and ileum, and feces were collected to estimate nutrient digestibility using Cr2O3 and TiO2 as external flow markers. Then, gas exchanges were measured in respiration chambers. There was no CH4 mitigating effect of MET. Nitrate reduced CH4 production (g/d), yield (g/kg DMI), and intensity (g/kg ECM) by 19.5, 11.9, and 17.2 %, respectively, whereas H2 yield (g/kg DMI) was increased by 261 %. Ruminal redox value was decreased by nitrate, and individual rumen volatile fatty acid proportions reflected a more reduced rumen environment although propionate proportions decreased. Nutrient digestibility was not affected by nitrate although microbial CP efficiency (g of microbial CP/kg true rumen digested organic matter) was decreased. Supplementing fumaric acid in combination with nitrate decreased H2 production by 26.8 % compared to nitrate fed cows, and this was associated with increased propionate concentrations. However, there was no effect on H2 emission when corrected for dry matter intake (DMI) or energy-corrected milk (ECM) yield. There were no effects of any of the treatments on DMI or ECM yield. In conclusion, the results demonstrated a CH4 mitigating effect of nitrate supplementation resulting in increased H2 emission. The effects on nutrient digestibility and rumen fermentation were minor. Fumaric acid supplementation redirected some H2 to propionate, although the efficiency was small. Supplementation of methanotrophic bacteria did not suppress CH4 emission.

Evaluating a process‐guided deep learning approach for predicting dissolved oxygen in streams

AbstractDissolved oxygen (DO) is a critical water quality constituent that governs habitat suitability for aquatic biota, biogeochemical reactions and solubility of metals in streams. Recently introduced high‐frequency sensors have increased our ability to measure DO, but we still lack the capacity to understand and predict DO concentrations at high spatial resolutions or in unmonitored locations. Machine learning (ML) has been a commonly used approach for modelling DO, however, conventional ML models have no representation of the limnological processes governing DO dynamics. Here we implement and evaluate two process‐guided deep learning (PGDL) approaches for predicting daily minimum, mean and maximum DO concentrations in rivers from the Delaware River Basin, USA. In both cases, a multi‐task approach was taken in which the PGDL models predicted stream metabolism and gas exchange rates in addition to the DO concentrations themselves. Our results showed that for these sites, the PGDL approaches did not improve upon baseline predictions in temporal and spatially similar holdout experiments. One of the approaches did, however, improve predictions when applied to spatially dissimilar sites. Although this particular PGDL approach did not improve predictive accuracy in most cases, our results suggest that process guidance, perhaps a more constrained approach, could benefit a data‐driven DO model.

Spatially structured exchange of metabolites enhances bacterial survival and resilience in biofilms

Biofilm formation enhances bacterial survival and antibiotic tolerance, but the underlying mechanisms are incompletely understood. Here, we show that biofilm growth is accompanied by a reduction in bacterial energy metabolism and membrane potential, together with metabolic exchanges between the inner and outer regions in biofilms. More specifically, nutrient-starved cells in the interior supply amino acids to cells in the periphery, while peripheral cells experience a decrease in membrane potential and provide fatty acids to interior cells. Fatty acids facilitate the repair of starvation-induced membrane damage in inner cells and enhance their survival in the presence of antibiotics. Thus, metabolic exchanges between inner and outer cells contribute to survival of the nutrient-starved inner cells and contribute to antibiotic tolerance within the biofilm.

Rescue of pyrimidine-defective Pseudomonas aeruginosa through metabolic complementation.

This research holds crucial implications for combating chronic infections, where multiple pathogens coexist and interact within the same environment. By uncovering the dynamic exchange of pyrimidines between Pseudomonas aeruginosa (PA) and Staphylococcus aureus (SA), our study reveals a previously unrecognized aspect of interspecies cooperation. The observed enhanced growth of a pyrimidine-deficient PA strain when co-cultured with SA suggests potential avenues for understanding and disrupting bacterial metabolic interdependencies in chronic infection settings. Furthermore, our findings highlight the mechanisms involved in metabolic exchange, emphasizing the importance of cell-to-cell contact. This research explored essential metabolic interactions to address the challenges posed by difficult-to-treat chronic infections.

Space, the final frontier: The spatial component of phytoplankton-bacterial interactions.

Microscale interactions between marine phytoplankton and bacteria shape the microenvironment of individual cells, impacting their physiology and ultimately influencing global-scale biogeochemical processes like carbon and nutrient cycling. In dilute environments such as the ocean water column, metabolic exchange between microorganisms likely requires close proximity between partners. However, the biological strategies to achieve this physical proximity remain an understudied aspect of phytoplankton-bacterial associations. Understanding the mechanisms by which these microorganisms establish and sustain spatial relationships and the extent to which spatial proximity is necessary for interactions to occur, is critical to learning how spatial associations influence the ecology of phytoplankton and bacterial communities. Here, we provide an overview of current knowledge on the role of space in shaping interactions among ocean microorganisms, encompassing behavioural and metabolic evidence. We propose that characterising phytoplankton-bacterial interactions from a spatial perspective can contribute to a mechanistic understanding of the establishment and maintenance of these associations and, consequently, an enhanced ability to predict the impact of microscale processes on ecosystem-wide phenomena.

Symbiodiniaceae and Ruegeria sp. Co-Cultivation to Enhance Nutrient Exchanges in Coral Holobiont.

The symbiotic relationship between corals and their associated microorganisms is crucial for the health of coral reef eco-environmental systems. Recently, there has been a growing interest in unraveling how the manipulation of symbiont nutrient cycling affects the stress tolerance in the holobiont of coral reefs. However, most studies have primarily focused on coral-Symbiodiniaceae-bacterial interactions as a whole, neglecting the interactions between Symbiodiniaceae and bacteria, which remain largely unexplored. In this study, we proposed a hypothesis that there exists an inner symbiotic loop of Symbiodiniaceae and bacteria within the coral symbiotic loop. We conducted experiments to demonstrate how metabolic exchanges between Symbiodiniaceae and bacteria facilitate the nutritional supply necessary for cellular growth. It was seen that the beneficial bacterium, Ruegeria sp., supplied a nitrogen source to the Symbiodiniaceae strain Durusdinium sp., allowing this dinoflagellate to thrive in a nitrogen-free medium. The Ruegeria sp.-Durusdinium sp. interaction was confirmed through 15N-stable isotope probing-single cell Raman spectroscopy, in which 15N infiltrated into the bacterial cells for intracellular metabolism, and eventually the labeled nitrogen source was traced within the macromolecules of Symbiodiniaceae cells. The investigation into Symbiodiniaceae loop interactions validates our hypothesis and contributes to a comprehensive understanding of the intricate coral holobiont. These findings have the potential to enhance the health of coral reefs in the face of global climate change.

Paired metabolomics and volatilomics provides insight into transient high light stress response mechanisms of the coral Montipora mollis

The coral holobiont is underpinned by complex metabolic exchanges between different symbiotic partners, which are impacted by environmental stressors. The chemical diversity of the compounds produced by the holobiont is high and includes primary and secondary metabolites, as well as volatiles. However, metabolites and volatiles have only been characterised in isolation so far. Here, we applied a paired metabolomic-volatilomic approach to characterise holistically the chemical response of the holobiont under stress. Montipora mollis fragments were subjected to high-light stress (8-fold higher than the controls) for 30 min. Photosystem II (PSII) photochemical efficiency values were 7-fold higher in control versus treatment corals immediately following high-light exposure, but returned to pre-stress levels after 30 min of recovery. Under high-light stress, we identified an increase in carbohydrates (> 5-fold increase in arabinose and fructose) and saturated fatty acids (7-fold increase in myristic and oleic acid), together with a decrease in fatty acid derivatives in both metabolites and volatiles (e.g., 80% decrease in oleamide and nonanal), and other antioxidants (~ 85% decrease in sorbitol and galactitol). These changes suggest short-term light stress induces oxidative stress. Correlation analysis between volatiles and metabolites identified positive links between sorbitol, galactitol, six other metabolites and 11 volatiles, with four of these compounds previously identified as antioxidants. This suggests that these 19 compounds may be related and share similar functions. Taken together, our findings demonstrate how paired metabolomics-volatilomics may illuminate broader metabolic shifts occurring under stress and identify linkages between uncharacterised compounds to putatively determine their functions.

Hundreds of antimicrobial peptides create a selective barrier for insect gut symbionts

The spatial organization of gut microbiota is crucial for the functioning of the gut ecosystem, although the mechanisms that organize gut bacterial communities in microhabitats are only partially understood. The gut of the insect Riptortus pedestris has a characteristic microbiota biogeography with a multispecies community in the anterior midgut and a monospecific bacterial population in the posterior midgut. We show that the posterior midgut region produces massively hundreds of specific antimicrobial peptides (AMPs), the Crypt-specific Cysteine-Rich peptides (CCRs) that have membrane-damaging antimicrobial activity against diverse bacteria but posterior midgut symbionts have elevated resistance. We determined by transposon-sequencing the genetic repertoire in the symbiont Caballeronia insecticola to manage CCR stress, identifying different independent pathways, including AMP-resistance pathways unrelated to known membrane homeostasis functions as well as cell envelope functions. Mutants in the corresponding genes have reduced capacity to colonize the posterior midgut, demonstrating that CCRs create a selective barrier and resistance is crucial in gut symbionts. Moreover, once established in the gut, the bacteria differentiate into a CCR-sensitive state, suggesting a second function of the CCR peptide arsenal in protecting the gut epithelia or mediating metabolic exchanges between the host and the gut symbionts. Our study highlights the evolution of an extreme diverse AMP family that likely contributes to establish and control the gut microbiota.

Endosymbiosis in trypanosomatids: The bacterium division depends on microtubule dynamism

Microtubules are components of the cytoskeleton that perform essential functions in eukaryotes, such as those related to shape change, motility and cell division. In this context some characteristics of these filaments are essential, such as polarity and dynamic instability. In trypanosomatids, microtubules are integral to ultrastructure organization, intracellular transport and mitotic processes. Some species of trypanosomatids co-evolve with a symbiotic bacterium in a mutualistic association that is marked by extensive metabolic exchanges and a coordinated division of the symbiont with other cellular structures, such as the nucleus and the kinetoplast. It is already established that the bacterium division is microtubule-dependent, so in this work, it was investigated whether the dynamism and remodeling of these filaments is capable of affecting the prokaryote division. To this purpose, Angomonas deanei was treated with Trichostatin A (TSA), a deacetylase inhibitor, and mutant cells for histone deacetylase 6 (HDAC6) were obtained by CRISPR-Cas9. A decrease in proliferation, an enhancement in tubulin acetylation, as well as morphological and ultrastructural changes, were observed in TSA-treated protozoa and mutant cells. In both cases, symbiont filamentation occurred, indicating that prokaryote cell division is dependent on microtubule dynamism.