Interspecies Interactions Research Articles

Learning beyond-pairwise interactions enables the bottom–up prediction of microbial community structure

Understanding the assembly of multispecies microbial communities represents a significant challenge in ecology and has wide applications in agriculture, wastewater treatment, and human healthcare domains. Traditionally, studies on the microbial community assembly focused on analyzing pairwise relationships among species; however, neglecting higher-order interactions, i.e., the change of pairwise relationships in the community context, may lead to substantial deviation from reality. Herein, we have proposed a simple framework that incorporates higher-order interactions into a bottom-up prediction of the microbial community assembly and examined its accuracy using a seven-member synthetic bacterial community on a host plant, duckweed. Although the synthetic community exhibited emergent properties that cannot be predicted from pairwise coculturing results, our results demonstrated that incorporating information from three-member combinations allows the acceptable prediction of the community structure and actual interaction forces within it. This reflects that the occurrence of higher-order effects follows consistent patterns, which can be predicted even from trio combinations, the smallest unit of higher-order interactions. These results highlight the possibility of predicting, explaining, and understanding the microbial community structure from the bottom-up by learning interspecies interactions from simple beyond-pairwise combinations.

Type VI secretion system drives bacterial diversity and functions in multispecies biofilms

Type VI secretion system (T6SS) plays an essential role in interspecies interactions and provides an advantage for a strain with T6SS in multispecies biofilms. However, how T6SS drives the bacterial community structure and functions in multispecies biofilms still needs to be determined. Using gene deletion and Illumina sequencing technique, we estimated bacterial community responses in multispecies biofilms to T6SS by introducing T6SS-containing Pseudomonas putida KT2440. Results showed that the niche structure shifts of multispecies biofilms were remarkably higher in the presence of T6SS than in the absence of T6SS. The presence of T6SS significantly drove the variation in microbial composition, reduced the alpha-diversity of bacterial communities in multispecies biofilms, and separately decreased and increased the relative abundance of Proteobacteria and Bacteroidota. Co-occurrence network analysis with inferred putative bacterial interactions indicated that P. putida KT2440 mainly displayed strong negative associations with the genera of Psychrobacter, Cellvibrio, Stenotrophomonas, and Brevundimonas. Moreover, the function redundancy index of the bacterial community was strikingly higher in the presence of T6SS than in the absence of T6SS, regardless of whether relative abundances of bacterial taxa were inhibited or promoted. Remarkably, the increased metabolic network similarity with T6SS-containing P. putida KT2440 could enhance the antibacterial activity of P. putida KT2440 on other bacterial taxa. Our findings extend knowledge of microbial adaptation strategies to potential bacterial weapons and could contribute to predicting biodiversity loss and change in ecological functions caused by T6SS.

Chemical oscillation in ultracold chemistry

We demonstrate the occurrence of oscillatory reactions in the ultra-cold chemistry of atom-molecular Bose-Einstein condensate. Nonlinear oscillations in the mean-field dynamics occur for a specific range of elliptic modulus, giving rise to both in- and out-phase modulations in the atom-molecule population density. The reaction front velocity is found to be controlled by photoassociation, which also regulates the condensate density. Two distinct pairs of in-phase bright localized gap solitons are found as exact solutions, the existence of one of which necessarily requires a background. Cnoidal atomic density waves along with a plane-wave molecular motion are observed in both attractive and repulsive domains. The role of intra- and inter-species interactions on both existence and stability is explicated in the presence of photoassociation.

Interspecies metabolite transfer fuels the methionine metabolism of Fusobacterium nucleatum to stimulate volatile methyl mercaptan production.

The major oral odor compound methyl mercaptan (CH3SH) is strongly associated with halitosis and periodontitis. CH3SH production stems from the metabolism of polymicrobial communities in periodontal pockets and on the tongue dorsum. However, understanding of CH3SH-producing oral bacteria and their interactions is limited. This study aimed to investigate CH3SH production by major oral bacteria and the impact of interspecies interactions on its generation. Using a newly constructed large-volume anaerobic noncontact coculture system, Fusobacterium nucleatum was found to be a potent producer of CH3SH, with that production stimulated by metabolic interactions with Streptococcus gordonii, an early dental plaque colonizer. Furthermore, analysis of extracellular amino acids using an S. gordonii arginine-ornithine antiporter (ArcD) mutant demonstrated that ornithine excreted from S. gordonii is a key contributor to increased CH3SH production by F. nucleatum. Further study with 13C, 15N-methionine, as well as gene expression analysis, revealed that ornithine secreted by S. gordonii increased the demand for methionine through accelerated polyamine synthesis by F. nucleatum, leading to elevated methionine pathway activity and CH3SH production. Collectively, these findings suggest that interaction between S. gordonii and F. nucleatum plays a key role in CH3SH production, providing a new insight into the mechanism of CH3SH generation in oral microbial communities. A better understanding of the underlying interactions among oral bacteria involved in CH3SH generation can lead to the development of more appropriate prophylactic approaches to treat halitosis and periodontitis. An intervention approach like selectively disrupting this interspecies network could also offer a powerful therapeutic strategy.IMPORTANCEHalitosis can have a significant impact on the social life of affected individuals. Among oral odor compounds, CH3SH has a low olfactory threshold and halitosis is a result of its production. Recently, there has been a growing interest in the collective properties of oral polymicrobial communities, regarded as important for the development of oral diseases, which are shaped by physical and metabolic interactions among community participants. However, it has yet to be investigated whether interspecies interactions have an impact on the production of volatile compounds, leading to the development of halitosis. The present findings provide mechanistic insights indicating that ornithine, a metabolite excreted by Streptococcus gordonii, promotes polyamine synthesis by Fusobacterium nucleatum, resulting in a compensatory increase in demand for methionine, which results in elevated methionine pathway activity and CH3SH production. Elucidation of the mechanisms related to CH3SH production is expected to lead to the development of new strategies for managing halitosis.

Coupled-cluster theory for trapped bosonic mixtures.

We develop a coupled-cluster theory for bosonic mixtures of binary species in external traps, providing a promising theoretical approach to demonstrate highly accurately the many-body physics of mixtures of Bose-Einstein condensates. The coupled-cluster wavefunction for the binary species is obtained when an exponential cluster operator eT, where T = T(1) + T(2) + T(12) and T(1) accounts for excitations in species-1, T(2) for excitations in species-2, and T(12) for combined excitations in both species, acts on the ground state configuration prepared by accumulating all bosons in a single orbital for each species. We have explicitly derived the working equations for bosonic mixtures by truncating the cluster operator up to the single and double excitations and using arbitrary sets of orthonormal orbitals for each of the species. Furthermore, the comparatively simplified version of the working equations are formulated using the Fock-like operators. Finally, using an exactly solvable many-body model for bosonic mixtures that exists in the literature allows us to implement and test the performance and accuracy of the coupled-cluster theory for situations with balanced as well as imbalanced boson numbers and for weak to moderately strong intra- and interspecies interaction strengths. The comparison between our computed results using coupled-cluster theory with the respective analytical exact results displays remarkable agreement exhibiting excellent success of the coupled-cluster theory for bosonic mixtures. All in all, the correlation exhaustive coupled-cluster theory shows encouraging results and could be a promising approach in paving the way for high-accuracy modeling of various bosonic mixture systems.

Interbacterial Biofilm Competition through a Suite of Secreted Metabolites.

Polymicrobial biofilms are ubiquitous, and the complex interspecies interactions within them are cryptic. We discovered the chemical foundation of antagonistic interactions in a model dual-species biofilm in which Pseudomonas aeruginosa inhibits the biofilm formation of Agrobacterium tumefaciens. Three known siderophores produced by P. aeruginosa (pyoverdine, pyochelin, and dihydroaeruginoic acid) were each capable of inhibiting biofilm formation. Surprisingly, a mutant that was incapable of producing these siderophores still secreted an antibiofilm metabolite. We discovered that this inhibitor was N5-formyl-N5-hydroxy-l-ornithine (fOHOrn)─a precursor in pyoverdine biosynthesis. Unlike the siderophores, this inhibitor did not appear to function via extracellular metal sequestration. In addition to this discovery, the compensatory overproduction of a new biofilm inhibitor illustrates the risk of pleiotropy in genetic knockout experiments. In total, this work lends new insight into the chemical nature of dual-species biofilm regulation and reveals a new naturally produced inhibitor of A. tumefaciens biofilm formation.

Engineered probiotic overcomes pathogen defences using signal interference and antibiotic production to treat infection in mice

Probiotic supplements are suggested to promote human health by preventing pathogen colonization. However, the mechanistic bases for their efficacy in vivo are largely uncharacterized. Here using metabolomics and bacterial genetics, we show that the human oral probiotic Streptococcus salivarius K12 (SAL) produces salivabactin, an antibiotic that effectively inhibits pathogenic Streptococcus pyogenes (GAS) in vitro and in mice. However, prophylactic dosing with SAL enhanced GAS colonization in mice and ex vivo in human saliva. We showed that, on co-colonization, GAS responds to a SAL intercellular peptide signal that controls SAL salivabactin production. GAS produces a secreted protease, SpeB, that targets SAL-derived salivaricins and enhances GAS survival. Using this knowledge, we re-engineered probiotic SAL to prevent signal eavesdropping by GAS and potentiate SAL antimicrobials. This engineered probiotic demonstrated superior efficacy in preventing GAS colonization in vivo. Our findings show that knowledge of interspecies interactions can identify antibiotic- and probiotic-based strategies to combat infection.

Does daily activity overlap of seven mesocarnivores vary based on human development?

Many species of wildlife alter their daily activity patterns in response to co-occurring species as well as the surrounding environment. Often smaller or subordinate species alter their activity patterns to avoid being active at the same time as larger, dominant species to avoid agonistic interactions. Human development can complicate interspecies interactions, as not all wildlife respond to human activity in the same manner. While some species may change the timing of their activity to avoid being active when humans are, others may be unaffected or may benefit from being active at the same time as humans to reduce predation risk or competition. To further explore these patterns, we used data from a coordinated national camera-trapping program (Snapshot USA) to explore how the activity patterns and temporal activity overlap of a suite of seven widely co-occurring mammalian mesocarnivores varied along a gradient of human development. Our focal species ranged in size from the large and often dominant coyote (Canis latrans) to the much smaller and subordinate Virginia opossum (Didelphis virginiana). Some species changed their activity based on surrounding human development. Coyotes were most active at night in areas of high and medium human development. Red fox (Vulpes vulpes) were more active at dusk in areas of high development relative to areas of low or medium development. However, because most species were primarily nocturnal regardless of human development, temporal activity overlap was high between all species. Only opossum and raccoon (Procyon lotor) showed changes in activity overlap with high overlap in areas of low development compared to areas of moderate development. Although we found that coyotes and red fox altered their activity patterns in response to human development, our results showed that competitive and predatory pressures between these seven widespread generalist species were insufficient to cause them to substantially alter their activity patterns.

Agonistic interactions initiated by adult bottlenose dolphins on Antillean manatee calves in the Caribbean Sea.

The dynamics and drivers of inter-species interactions in the wild are poorly understood, particularly those involving social animal species. Inter-species interactions between cetaceans and sirenians have rarely been documented and investigated. Here, we report 10 cases of interaction initiated by adult bottlenose dolphins (Tursiops truncatus) towards Antillean manatee (Trichechus manatus manatus). Interactions were documented through behavioral observations in the wild (n = 7) and from the examination of orphaned calves (i.e., tooth rake marks on their body; n = 4) that entered a rehabilitation facility, one individual both observed interacting with dolphins and found stranded with bite marks. Bottlenose dolphins were observed interacting with orphan manatee calves and with mother-calf pairs, exhibiting agonistic behavior (n = 2), affiliative or neutral behaviors (n = 1), but the behavioral contexts of these interactions remain unclear in most cases (n = 7). Information on stranded individuals was collected from four calves (of 13 examined calves) recovered in poor condition with bottlenose dolphin tooth rakes and bite wounds on their bodies, one of which died. Injury from bite wounds varied in extent and severity, ranging from superficial scratches leaving rake marks to deep lacerations. Our findings suggest the regular occurrence of agonistic behaviors initiated by adult bottlenose dolphins and directed toward manatee calves. However, the drivers of these interactions remain unknown and need to be further investigated.

Entomopathogenic pseudomonads can share an insect host with entomopathogenic nematodes and their mutualistic bacteria.

A promising strategy to overcome limitations in biological control of insect pests is the combined application of entomopathogenic pseudomonads (EPPs) and nematodes (EPNs) associated with mutualistic bacteria (NABs). Yet, little is known about interspecies interactions such as competition, coexistence, or even cooperation between these entomopathogens when they infect the same insect host. We investigated the dynamics of bacteria-bacteria interactions between the EPP Pseudomonas protegens CHA0 and the NAB Xenorhabdus bovienii SM5 isolated from the EPN Steinernema feltiae RS5. Bacterial populations were assessed over time in experimental systems of increasing complexity. In vitro, SM5 was outcompeted when CHA0 reached a certain cell density, resulting in the collapse of the SM5 population. In contrast, both bacteria were able to coexist upon haemolymph-injection into Galleria mellonella larvae, as found for three further EPP-NAB combinations. Finally, both bacteria were administered by natural infection routes i.e. orally for CHA0 and nematode-vectored for SM5 resulting in the addition of RS5 to the system. This did not alter bacterial coexistence nor did the presence of the EPP affect nematode reproductive success or progeny virulence. CHA0 benefited from RS5, probably by exploiting access routes formed by the nematodes penetrating the larval gut epithelium. Our results indicate that EPPs are able to share an insect host with EPNs and their mutualistic bacteria without major negative effects on the reproduction of any of the three entomopathogens or the fitness of the nematodes. This suggests that their combination is a promising strategy for biological insect pest control.

Hypervirulent Klebsiella pneumoniae employs genomic island encoded toxins against bacterial competitors in the gut.

The hypervirulent lineages of Klebsiella pneumoniae (HvKp) cause invasive infections such as Klebsiella-liver abscess. Invasive infection often occurs after initial colonization of the host gastrointestinal tract by HvKp. Over 80% of HvKp isolates belong to the clonal group 23 sublineage I that has acquired genomic islands (GIs) GIE492 and ICEKp10. Our analysis of 12 361K. pneumoniae genomes revealed that GIs GIE492 and ICEKp10 are co-associated with the CG23-I and CG10118 HvKp lineages. GIE492 and ICEKp10 enable HvKp to make a functional bacteriocin microcin E492 (mccE492) and the genotoxin colibactin, respectively. We discovered that GIE492 and ICEKp10 play cooperative roles and enhance gastrointestinal colonization by HvKp. Colibactin is the primary driver of this effect, modifying gut microbiome diversity. Our in vitro assays demonstrate that colibactin and mccE492 kill or inhibit a range of Gram-negative Klebsiella species and Escherichia coli strains, including Gram-positive bacteria, sometimes cooperatively. Moreover, mccE492 and colibactin kill human anaerobic gut commensals that are similar to the taxa found altered by colibactin in the mouse intestines. Our findings suggest that GIs GIE492 and ICEKp10 enable HvKp to kill several commensal bacterial taxa during interspecies interactions in the gut. Thus, acquisition of GIE492 and ICEKp10 could enable better carriage in host populations and explain the dominance of the CG23-I HvKp lineage.

Constructions of quorum sensing signaling network for activated sludge microbial community.

In wastewater treatment systems, the interactions among various microbes based on chemical signals, namely quorum sensing (QS), play critical roles in influencing microbial structure and function. However, it is challenging to understand the QS-controlled behaviors and the underlying mechanisms in complex microbial communities. In this study, we constructed a QS signaling network, providing insights into the intra- and interspecies interactions of activated sludge microbial communities based on diverse QS signal molecules. Our research underscores the role of diffusible signal factors in both intra- and interspecies communication among activated sludge microorganisms, and signal molecules commonly considered to mediate intraspecies communication may also participate in interspecies communication. QS signaling molecules play an important role as communal resources among the entire microbial group. The communication network within the microbial community is highly redundant, significantly contributing to the stability of natural microbial systems. This work contributes to the establishment of QS signaling network for activated sludge microbial communities, which may complement metabolic exchanges in explaining activated sludge microbial community structure and may help with a variety of future applications, such as making the dynamics and resilience of highly complex ecosystems more predictable.

Probing interspecies metabolic interactions within a synthetic binary microbiome using genome-scale modeling.

Aim: Metabolic interactions within a microbial community play a key role in determining the structure, function, and composition of the community. However, due to the complexity and intractability of natural microbiomes, limited knowledge is available on interspecies interactions within a community. In this work, using a binary synthetic microbiome, a methanotroph-photoautotroph (M-P) coculture, as the model system, we examined different genome-scale metabolic modeling (GEM) approaches to gain a better understanding of the metabolic interactions within the coculture, how they contribute to the enhanced growth observed in the coculture, and how they evolve over time. Methods: Using batch growth data of the model M-P coculture, we compared three GEM approaches for microbial communities. Two of the methods are existing approaches: SteadyCom, a steady state GEM, and dynamic flux balance analysis (DFBA) Lab, a dynamic GEM. We also proposed an improved dynamic GEM approach, DynamiCom, for the M-P coculture. Results: SteadyCom can predict the metabolic interactions within the coculture but not their dynamic evolutions; DFBA Lab can predict the dynamics of the coculture but cannot identify interspecies interactions. DynamiCom was able to identify the cross-fed metabolite within the coculture, as well as predict the evolution of the interspecies interactions over time. Conclusion: A new dynamic GEM approach, DynamiCom, was developed for a model M-P coculture. Constrained by the predictions from a validated kinetic model, DynamiCom consistently predicted the top metabolites being exchanged in the M-P coculture, as well as the establishment of the mutualistic N-exchange between the methanotroph and cyanobacteria. The interspecies interactions and their dynamic evolution predicted by DynamiCom are supported by ample evidence in the literature on methanotroph, cyanobacteria, and other cyanobacteria-heterotroph cocultures.

Gene Ontology and pathway enrichment analysis of the upregulated biological Proteins in Single-Line Transgenic Pod Borer Resistant Cowpea

Aim: This research seeks to clarify the proteomic alterations linked to PBR cowpea in comparison to its non-transgenic isoline. Gene Ontology and pathway enrichment analyses were conducted to pinpoint critical biological processes, molecular functions, and cellular components that exhibit upregulation in the transgenic cowpea. Methods: Seeds from both cowpea lines were finely ground, and proteins were extracted utilizing Tris-EDTA and LDS buffers. The proteins were then separated and identified through mass spectrometry (MS), with peptide sequences being matched against protein sequence databases. Results: The enrichment analysis indicated upregulation in pathways associated with pollen tube adhesion, cell-cell adhesion, and biological processes pertinent to both intraspecies and interspecies interactions with no significant difference. Importantly, pathways linked to defense mechanisms against fungi and other organisms were notably enriched, suggesting an enhanced ability to detect and respond to biotic stresses. The results of this study does not underscore any substantial proteomic modifications in the transgenic cowpea. Conclusion: These findings offer valuable targets for genetic engineering and breeding initiatives aimed at creating cowpea varieties with enhanced pest resistance, thus improving crop yield and sustainability. Future investigations should concentrate on validating these proteins and pathways through functional assays and examining their roles in resistance under field conditions.

Host‐, Environment‐, or Human‐Related Effects Drive Interspecies Interactions in an Animal Tuberculosis Multi‐Host Community Depending on the Host and Season

In many Mediterranean ecosystems, animal tuberculosis (TB), caused by Mycobacterium bovis, is maintained by multi‐host communities in which cattle and different wildlife species establish interaction networks contributing to M. bovis transmission and persistence. Most studies have addressed wildlife–cattle disease‐relevant interactions, focusing on reservoir hosts, while disregarding the potential contribution of the so‐called accidental hosts and/or neglecting wildlife–wildlife interactions. In this work, we aimed to characterise interspecies interactions in an endemic TB risk area and identify the ecological drivers of interaction patterns regardless of the pre‐attributed role of host species on TB epidemiology. For that purpose, spatial–temporal indirect interactions between wildlife mammals and cattle, and between different wildlife species, were investigated through camera trapping. Second, five ecological hypotheses potentially driving species pair interactions in the wet and dry seasons were tested covering water and control sites: human presence (H1), landscape composition (H2), topography (H3), weather (H4), and natural food and water resources (H5). Wild boar (Sus scrofa), red deer (Cervus elaphus), and red fox (Vulpes vulpes) were the wildlife species mostly involved in indirect interactions. We found that indirect wildlife–cattle interactions were more frequent than wildlife interactions and, for certain species pairs, interaction rates were higher in the wet season in both wildlife–cattle and wildlife groups. Natural food and water resources (H5) was the most supported hypothesis that influenced the abundance of wildlife–cattle interactions, with positive effects during the dry season and negative effects during the wet season. In contrast, the abundance of indirect interactions between wildlife species was mainly supported by the human disturbance hypothesis (H1), with negative effects exerted on the dry season and variable effects on the wet season. Other tested hypotheses also influenced wildlife–cattle and wildlife–wildlife interactions, depending on the season and host species. These results highlight that indirect interactions, and thus conditions potentially favouring the transmission of M. bovis in shared environments, are determined by different ecological backgrounds.

Pavement ant extract is a chemotaxis repellent for C. elegans.

Ant behavior relies on a collection of natural products, from following trail pheromones during foraging to warding off potential predators. How nervous systems sense these compounds to initiate a behavioral response remains unclear. Here, we used Caenorhabditis elegans chemotaxis assays to investigate how ant compounds are detected by heterospecific nervous systems. We found that C. elegans avoid extracts of the pavement ant ( Tetramorium immigrans ) and either osm-9 or tax-4 ion channels are required for this response. These experiments were conducted in an undergraduate laboratory course, demonstrating that new insights into interspecies interactions can be generated through genuine research experiences in a classroom setting.

The discourse of ecological proximity in The Famished Road and Wizard of the Crow: A postcolonial ecocritical analysis

This study explicates the discourse of ecological proximity embedded in two postcolonial African novels. It employed ecocritical theory informed by posthumanism, ‘aesthetics of proximity’ (Iheka, 2018) and postcolonial ecocriticism as critical venture to undertake analysis. The novels targeted in the study, The Famished Road, and Wizard of the Crow, were purposely selected owing to their thematic preoccupation and stylistic convergence pertinent to proximity of ecological entities. The study reveals that both novels present discourses that critique hierarchy in the relationship among ecological entities and promote harmonious interaction. The proximity discourses are revealed through foregrounding ecological diversity in portrayal of settings, depicting harmony in interspecies interaction, conferring power of agency to the nonhuman entities, and through blurring anthropocentric boundaries. Through distributing agency to non-human nature and endowing human figures the capacity to step out of anthropocentric confines, the novelists have foregrounded a discourse of proximity among ecological entities in their narratives.

Transport connectivity of the organic production zone at the regional level

The paper presents an assessment of the development prospects and transport accessibility of organic production zones in the regions of Siberia based on data from state information systems of the Russian Federation and open information systems of commercial digital aggregators. The assessment was carried out with the use of technologies and analytics of the ArcGIS 10.6 program. The Novosibirsk, Omsk, Barnaul, and Tomsk agglomerations have concentrated clusters of investment projects for the processing of agricultural products. The eastern part of the macrozone is characterized by more focal investment activity, the largest of which is the eastern agglomeration of Krasnoyarsk. Investment projects in Kuzbass and the Irkutsk region are dispersed along the main transport routes. The republics of Altai and Tyva and the northern municipalities of the Tomsk and Irkutsk regions remain on the periphery of active transport contacts. Inside the highway part of the region there are areas of low efficiency in the use of transport potential, which makes it possible to increase the investment attractiveness of individual agricultural locations based on a combination of interspecies interaction.

Total substitution and partial modification of the set of non-ribosomal peptide synthetases clusters lead to pyoverdine diversity in the Pseudomonas fluorescens complex.

Pyoverdines are high affinity siderophores produced by most Pseudomonas with a wide role in microbial interspecies interactions. They are primarily composed of a conserved chromophore moiety, an acyl side chain and a peptide backbone which may be highly variable among strains. Upon ferric iron sequestration, pyoverdines are internalized through specialized receptors. The peptide precursor of pyoverdine, termed ferribactin, is synthesized by a set of non-ribosomal peptide synthetase (NRPS) enzymes and further modified by tailoring enzymes. While PvdL, the NRPS responsible for the synthesis of the peptide moiety that derives into the chromophore is conserved, the NRPSs for the peptide backbone are different across fluorescent Pseudomonas. Although the variation of pyoverdine is a widely recognized characteristic within the genus, the evolutionary events associated with the diversity and distribution of this trait remain mostly unknown. This study analyzed the NRPSs clusters for the biosynthesis of the peptide backbone of ferribactin in the genomes of a representative subset of strains of the Pseudomonas fluorescens complex. Bioinformatic analysis of the specificity of adenylation domains of the NRPSs allowed the prediction of 30 different pyoverdine variants. Phylogenetic reconstruction and mapping of the NRPS clusters pinpointed two different general levels of modifications. In the first level, a complete replacement of the set of NRPRs by horizontal transfer occurs. In the second level, the original set of NRPSs is modified through different mechanisms, including partial substitution of the NRPS genes by horizontal transfer, adenylation domain specificity change or NRPS accessory domain gain/loss.

Construction and characterization of stable multi-species biofilms formed by nine core gut bacteria on wheat fiber.

Microbial aggregation mainly occurs on the intestinal epithelium, mucosal layer and undigested food particles in the gastrointestinal tract (GIT). Undigested food particles are usually insoluble dietary fiber (IDF), which can be easily obtained through daily diet, but there are few studies investigating whether the gut bacteria adhering to undigested food particles can form multi-species biofilms. In this study, we prepared mono- and multi-species biofilms using 18 core gut bacteria via a dynamic fermentation method, and it was found that multi-species composed of nine core gut bacteria (M9) showed the best biofilm formation ability. Cell counts of the nine bacteria in multi-species biofilms were 9.36, 11.85, 10.17, 9.93, 12.88, 11.39, 10.089, 9.06, and 13.21 Log10 CFU mL-1. M9 was tightly connected and regularly stacked on wheat fiber and had larger particle sizes than mono-species biofilms. M9 retained biofilm formation ability under pH and bile salt stresses. A human feces invasion experiment demonstrated that M9 can stably adhere to wheat fiber under the interference of complex gut bacteria, and the M9 multi-species biofilm had positions that can be filled by various gut bacteria. Metabolome results indicated that the M9 multi-species biofilm had more metabolic productions and more complex interspecies interactions than mono-species biofilms. This study provides a dynamic fermentation method to prepare multi-species biofilms on wheat fiber in vitro. It will also offer a research basis for clarifying whether gut bacteria can utilize IDF to form biofilm structures in vivo and the possible interspecific interactions and physiological functions of bacteria in biofilms.