Collective motion of bacteria in two dimensions

Collective motion can be observed in biological systems over a wide range of length scales, from large animals to bacteria. Collective motion is thought to confer an advantage for defense and adaptation. A central question in the study of biological collective motion is how the traits of individuals give rise to the emergent behavior at population level. This question is relevant to the dynamics of general self-propelled particle systems, biological self-organization, and active fluids. Bacteria provide a tractable system to address this question, because bacteria are simple and their behavior is relatively easy to control. In this mini review we will focus on a special form of bacterial collective motion, i.e., bacterial swarming in two dimensions. We will introduce some organization principles known in bacterial swarming and discuss potential means of controlling its dynamics. The simplicity and controllability of 2D bacterial behavior during swarming would allow experimental examination of theory predictions on general collective motion.     

Characterization of swarming motility in Citrobacter freundii

Bacterial swarming motility is a flagella-dependent translocation on the surface environment. It has received extensive attention as a population behavior involving numerous genes. Here, we report that Citrobacter freundii, an opportunistic pathogen, exhibits swarming movement on a solid medium surface with appropriate agar concentration. The swarming behavior of C. freundii was described in detail. Insertional mutagenesis with transposon Mini-Tn5 was carried out to discover genetic determinants related to the swarming of C. freundii. A number of swarming genes were identified, among which flhD, motA, motB, wzx, rfaL, rfaJ, rfbX, rfaG, rcsD, rcsC, gshB, fabF, dam, pgi, and rssB have been characterized previously in other species. In mutants related to lipopolysaccharide synthesis and RcsCDB signal system, a propensity to form poorly motile bacterial aggregates on the agar surface was observed. The aggregates hampered bacterial surface migration. In several mutants, the insertion sites were identified to be in the ORF of yqhC, yeeZ, CKO_03941, glgC, and ttrA, which have never been shown to be involved in swarming. Our results revealed several novel characteristics of swarming motility in C. freundii which are worthy of further study.     

Genetic Determinants of Swarming in Rhizobium etli

Swarming motility is considered to be a social phenomenon that enables groups of bacteria to move coordinately atop solid surfaces. The differentiated swarmer cell population is embedded in an extracellular slime layer, and the phenomenon has previously been linked with biofilm formation and virulence. The gram-negative nitrogen-fixing soil bacterium Rhizobium etli CNPAF512 was previously shown to display swarming behavior on soft agar plates. In a search for novel genetic determinants of swarming, a detailed analysis of the swarming behavior of 700 miniTn5 mutants of R. etli was performed. Twenty-four mutants defective in swarming or displaying abnormal swarming patterns were identified and could be divided into three groups based on their swarming pattern. Fourteen mutants were completely swarming deficient, five mutants showed an atypical swarming pattern with no completely smooth edge and local extrusions, and five mutants displayed an intermediate swarming phenotype. Sequence analysis of the targeted genes indicated that the mutants were likely affected in quorum-sensing, polysaccharide composition or export, motility, and amino acid and polyamines metabolism. Several of the identified mutants displayed a reduced symbiotic nitrogen fixation activity.     

Bees aren't the only ones: swarming in Gram-negative bacteria

Summary
Swarming is a form of active surface motility that is widespread among flagellated. Gram–negative bacteria. In the laboratory, growth of the bacteria on certain agar surfaces leads to induction of the differentiated swarmer-cell state. Swarmer cells are generally long and multinucleate, always hyperflagellated, and can move rapidly over the agar surface in a coordinated manner. Some swarm colonies exude large amounts of 'slime', which could be essential for promoting intimate cell–cell contacts during swarming. There is evidence that the differentiated swarmer-cell stage facilitates pathogenic associations with host tissue. Almost nothing is known about the molecular signalling mechanism of surface sensing. Increased viscosity appears to be sensed by several bacteria, but other environmental cues, specific to each bacterium, are also important. In organisms in which swarming motility has been studied in some detail, the chemotaxis system has been shown to play an important rote. The recent discovery of swarming motility in two genetically well-characterized organisms – Escherichia coli and Salmonella typhimurium – should lead to rapid progress in understanding this process.     

Swarming populations of<Emphasis Type="Italic">Salmonella</Emphasis> represent a unique physiological state coupled to multiple mechanisms of antibiotic resistance

Salmonella enterica serovar Typhimurium is capable of swarming over semi-solid surfaces. Although its swarming behavior shares many readily observable similarities with other swarming bacteria, the phenomenon remains somewhat of an enigma in this bacterium since some attributes skew away from the better characterized systems. Swarming is quite distinct from the classic swimming motility, as there is a prerequisite for cells to first undergo a morphological transformation into swarmer cells. In some organisms, swarming is controlled by quorum sensing, and in others, swarming has been shown to be coupled to increased expression of important virulence factors. Swarming in serovar Typhimurium is coupled to elevated resistance to a wide variety of structurally and functionally distinct classes of antimicrobial compounds. As serovar Typhimurium differentiates into swarm cells, thepmrHFIJKLM operon is up-regulated, resulting in a more positively charged LPS core. Furthermore, as swarm cells begin to de-differentiate, thepmr operon expression is down-regulated, rapidly reaching the levels observed in swim cells. This is one potential mechanism which confers swarm cells increased resistance to antibiotics such as the cationic antimicrobial peptides. However, additional mechanisms are likely associated with the cells in the swarm state that confer elevated resistance to such a broad spectrum of antimicrobial agents. Published: September 26, 2003     

Hybridization Hotspots at Bat Swarming Sites

During late summer and early autumn in temperate zones of the Northern Hemisphere, thousands of bats gather at caves, mainly for the purpose of mating. We demonstrated that this swarming behavior most probably leads not only to breeding among bats of the same species but also interbreeding between different species. Using 14 nuclear microsatellites and three different methods (the Bayesian assignment approaches of STRUCTURE and NEWHYBRIDS and a principal coordinate analysis of pairwise genetic distances), we analyzed 375 individuals belonging to three species of whiskered bats (genus Myotis) at swarming sites across their sympatric range in southern Poland. The overall hybridization rate varied from 3.2 to 7.2%. At the species level, depending on the method used, these values ranged from 2.1–4.6% in M. mystacinus and 3.0–3.7% in M. brandtii to 6.5–30.4% in M. alcathoe. Hybrids occurred in about half of the caves we studied. In all three species, the sex ratio of hybrids was biased towards males but the observed differences did not differ statistically from those noted at the population level. In our opinion, factors leading to the formation of these admixed individuals and their relatively high frequency are: i) swarming behaviour at swarming sites, where high numbers of bats belonging to several species meet; ii) male-biased sex ratio during the swarming period; iii) the fact that all these bats are generally polygynous. The highly different population sizes of different species at swarming sites may also play some role. Swarming sites may represent unique hybrid hotspots, which, as there are at least 2,000 caves in the Polish Carpathians alone, may occur on a massive scale not previously observed for any group of mammal species in the wild. Evidently, these sites should be treated as focal points for the conservation of biodiversity and evolutionary processes.     

Zooplankton swarms: characteristics,proximal cues and proposed advantages

This review focuses on monospecific swarms of four taxonomic groups of small crustaceans: three groups are marine copepods: oithonids (Oithona and Dioithona), Acartia species and Calanus species; and the fourth group includes freshwater cladoceran species in the Order Anomopoda. For each of these groups there is a substantial literature on swarming behavior from field studies and laboratory experiments. Swarming characteristics of each taxonomic group are reviewed, proximal cues for swarming are described, comparisons are made for proposed advantages of swarming, and future research directions are suggested. Swarming characteristics of Calanus spp. are distinctly different from those of the smaller crustaceans: the oithonids, acartiids and cladocerans. In a conceptual model proposed for the smaller crustaceans, swarming behavior is affected by their interactions with light cues, water currents and turbulence, behavior of their predators and prey, and abundance of other trophic levels.     

A Continuum Three-Zone Model for Swarms

We present a progression of three distinct three-zone, continuum models for swarm behavior based on social interactions with neighbors in order to explain simple coherent structures in popular biological models of aggregations. In continuum models, individuals are replaced with density and velocity functions. Individual behavior is modeled with convolutions acting within three interaction zones corresponding to repulsion, orientation, and attraction, respectively. We begin with a variable-speed first-order model in which the velocity depends directly on the interactions. Next, we present a variable-speed second-order model. Finally, we present a constant-speed second-order model that is coordinated with popular individual-based models. For all three models, linear stability analysis shows that the growth or decay of perturbations in an infinite, uniform swarm depends on the strength of attraction relative to repulsion and orientation. We verify that the continuum models predict the behavior of a swarm of individuals by comparing the linear stability results with an individual-based model that uses the same social interaction kernels. In some unstable regimes, we observe that the uniform state will evolve toward a radially symmetric attractor with a variable density. In other unstable regimes, we observe an incoherent swarming state.     

Swarm Site Fidelity in the Sex Role-Reversed Dance Fly Empis borealis

In the dance fly species Empis borealis (L.), females (1–40) gather to swarm at landmarks (swarm markers, like trees and bushes), and males carrying an insect prey visit these swarms for mating. We noticed earlier that some swarm sites were used for several years and that they appeared to be frequented by a similar number of swarming females in each year, although the numbers of females varied greatly among swarm sites and certain sites attracted more swarming individuals than others. To explore swarm site fidelity in this mating system, in 1993 we monitored the same swarm sites that we studied in 1989, addressing the questions, Would the same swarm sites still attract the same number of females and males after 4 years? and Why do some swarm sites attract more displaying females than others? The number of females swarming at the different markers in 1993 was approximately the same as 4 years earlier. Some of these swarm sites are known to have been used for 18 years. The swarm sites with the largest number of flies had a high sun exposure during the day and were found at coniferous swarm marker trees and in a mixed forest habitat. A swarm site with few females attending and with a low amount of insolation during the day can be predicted to be abandoned as a swarming site soon. Empis borealis swarm sites thus persist over many years and are attended by a similar number of individuals each year. To our knowledge, such site fidelity has not been demonstrated for any swarming insect species earlier.     

Epidemiology of honey bee parasites

The epidemiology of honey bee parasites has been somewhat neglected, but Lynn Royce and Philippe Rossignol describe their unique characteristics. Indeed, it appears that a parasite of social insects has in essence to adapt to two hosts: first, the individual worker within a colony, the numbers of which grow linearly and second, to the colony itself, the actual reproductive 'organism'. Transmission also has vertical and horizontal components. Analysis of tracheal mite populations in particular suggests that intracolony parasite levels are regulated by the swarming behavior of their hosts. Ironically, current and highly productive methods of honey bee management with movable hives curb swarming and may contribute to increasing the spread and the impact of some parasites. This insight may result in changing management practices to reduce the detrimental effects of bee parasites.     

Photophobic behavior of maize roots

Primary roots of young maize seedlings showed peculiar growth behavior when challenged by placing them on a slope, or if whole seedlings were turned upside down. Importantly, this behavior was dependent on the light conditions. If roots were placed on slopes in the dark, they performed “crawling” behavior and advanced rapidly up the slope. However, as soon as these roots were illuminated, their crawling movements along their horizontal paths slowed down, and instead tried to grow downwards along the gravity vector. A similar light-induced switch in the root behavior was observed when roots were inverted, by placing them in thin glass capillaries. As long as they were kept in the darkness, they showed rapid growth against the gravity vector. If illuminated, these inverted roots rapidly accomplished U-turns and grew down along the gravity vector, eventually escaping from the capillaries upon reaching their open ends. De-capped roots, although growing vigorously, did not display these light-induced photophobic growth responses. We can conclude that intact root cap is essential for the photophobic root behavior in maize.     

High gene diversity at swarming sites suggest hot spots for gene flow in the endangered Bechstein's bat

Our study shows that endangered Bechstein'sbats utilise distinct habitats at differentstages of their reproductive cycle, a findingthat has implications how habitat should beselected for preservation. Using nuclear andmitochondrial microsatellite DNA markers wecompared gene diversity of Bechstein's batswithin breeding colonies and at potentialmating sites. Bechstein's bat is one of themost threatened European bat species. Duringsummer it depends largely on mature deciduousforests. Females exhibit strict natalphilopatry. They form demographicallyindependent breeding colonies comprisingmaternally closely related bats. Males aresolitary. Like other temperate bats,Bechstein's bats swarm at the end of summer infront of caves. Because the sexes meet there,such swarming sites are potentially importantfor gene flow. Our genetic analyses reveal thatswarming sites have greater mitochondrial DNAgene diversity than colonies. Furthermore,field data on the phenology and reproductivestatus of several hundred individuals suggestthat Bechstein's bats mate during swarming. Incombination our field and genetic data showthat swarming sites provide the opportunity forgene flow among bats originating from differentcolonies. Therefore, we suggest that swarmingsites should be strictly protected to maintainthe observed high levels of gene flow amongcolonies.     

Characterization of Proteus mirabilis Precocious Swarming Mutants: Identification of rsbA, Encoding a Regulator of Swarming Behavior

Proteus mirabilis swarming behavior is characterized by the development of concentric rings of growth that are formed as cyclic events of swarmer cell differentiation, swarming migration, and cellular differentiation are repeated during colony translocation across a surface. This cycle produces the bull’s-eye colony often associated with cultures of P. mirabilis. How the cells communicate with one another to coordinate these perfectly synchronized rings is presently unknown. We report here the identification of a genetic locus that, when mutated, results in a precocious swarming phenotype. These mutants are defective in the temporal control of swarming migration and start swarming ca. 60 min sooner than wild-type cells. Unlike the wild type, precocious swarming mutants are also constitutive swarmer cells and swarm on minimal agar medium. The defects were found to be localized to a 5.4-kb locus on the P. mirabilis genome encoding RsbA (regulator of swarming behavior) and the P. mirabilis homologs to RcsB and RcsC. RsbA is homologous to membrane sensor histidine kinases of the two-component family of regulatory proteins, suggesting that RsbA may function as a sensor of environmental conditions required to initiate swarming migration. Introduction of a rsbA mutation back into the wild type via allelic-exchange mutagenesis reconstructed the precocious swarming phenotype, which could be complemented in trans by a plasmid-borne copy of rsbA. Overexpression of RsbA in wild-type cells resulted in precocious swarming, suggesting that RsbA may have both positive and negative functions in regulating swarming migration. A possible model to describe the role of RsbA in swarming migration is discussed.     

Smart swarms of bacteria-inspired agents with performance adaptable interactions

Collective navigation and swarming have been studied in animal groups, such as fish schools, bird flocks, bacteria, and slime molds. Computer modeling has shown that collective behavior of simple agents can result from simple interactions between the agents, which include short range repulsion, intermediate range alignment, and long range attraction. Here we study collective navigation of bacteria-inspired smart agents in complex terrains, with adaptive interactions that depend on performance. More specifically, each agent adjusts its interactions with the other agents according to its local environment--by decreasing the peers' influence while navigating in a beneficial direction, and increasing it otherwise. We show that inclusion of such performance dependent adaptable interactions significantly improves the collective swarming performance, leading to highly efficient navigation, especially in complex terrains. Notably, to afford such adaptable interactions, each modeled agent requires only simple computational capabilities with short-term memory, which can easily be implemented in simple swarming robots.     

The Yersinia enterocolitica motility master regulatory operon, flhDC, is required for flagellin production, swimming motility, and swarming motility

The ability to move over and colonize surface substrata has been linked to the formation of biofilms and to the virulence of some bacterial pathogens. Results from this study show that the gastrointestinal pathogen Yersinia enterocolitica can migrate over and colonize surfaces by swarming motility, a form of cooperative multicellular behavior. Immunoblot analysis and electron microscopy indicated that swarming motility is dependent on the same flagellum organelle that is required for swimming motility, which occurs in fluid environments. Furthermore, motility genes such as flgEF, flgMN, flhBA, and fliA, known to be required for the production of flagella, are essential for swarming motility. To begin to investigate how environmental signals are processed and integrated by Y. enterocolitica to stimulate the production of flagella and regulate these two forms of cell migration, the motility master regulatory operon, flhDC, was cloned. Mutations within flhDC completely abolished swimming motility, swarming motility, and flagellin production. DNA sequence analysis revealed that this locus is similar to motility master regulatory operons of other gram-negative bacteria. Genetic complementation and functional analysis of flhDC indicated that it is required for the production of flagella. When flhDC was expressed from an inducible ptac promoter, flagellin production was shown to be dependent on levels of flhDC expression. Phenotypically, induction of the ptac-flhDC fusion also corresponded to increased levels of both swimming and swarming motility.     

Extracellular proteolytic activity plays a central role in swarming motility in Bacillus subtilis

Natural isolates of Bacillus subtilis exhibit a robust multicellular behavior known as swarming. A form of motility, swarming is characterized by a rapid, coordinated progression of a bacterial population across a surface. As a collective bacterial process, swarming is often associated with biofilm formation and has been linked to virulence factor expression in pathogenic bacteria. While the swarming phenotype has been well documented for Bacillus species, an understanding of the molecular mechanisms responsible remains largely isolated to gram-negative bacteria. To better understand how swarming is controlled in members of the genus Bacillus, we investigated the effect of a series of gene deletions on swarm motility. Our analysis revealed that a strain deficient for the production of surfactin and extracellular proteolytic activity did not swarm or form biofilm. While it is known that surfactin, a lipoprotein surfactant, functions in swarming motility by reducing surface tension, this is the first report demonstrating that general extracellular protease activity also has an important function. These results not only help to define the factors involved in eliciting swarm migration but support the idea that swarming and biofilm formation may have overlapping control mechanisms.     

Effects of high concentrations of inorganic salts on swarming ability in fluorescent pseudomonas strains

We did tests using swarm plates, to examine the effects of various salts and their concentrations on the chemotaxis of fluorescent Pseudomonas strains. As a result, we found that the swarming ability of the Pseudomonas strains was inhibited by high concentrations of Ca2+. The growth of the strains was not affected at the high concentration of Ca2+, but the cells grown in swarm agar under the condition were extended in the filaments. Most of the cells had reached 10 microm to 40 microm in length. Such cell elongation was not observed with salts other than calcium salts. A significant correlation between the cell elongation and the decrease of swarming ability by the high concentration of Ca2+ was observed.     

Epr plays a key role in DegU-mediated swarming motility of Bacillus subtilis

Epr, a minor extracellular protease, is involved in the swarming motility of Bacillus subtilis . It does so by providing essential signals required for swarming. It has also been demonstrated that DegU is required for swarming and that it occurs at very low levels of DegU∼P and is inhibited at high levels of DegU∼P. In this study, we show that maximal epr expression is observed at very low concentrations of DegU∼P, whereas it is repressed at high DegU∼P. A parallel effect of DegU∼P levels on swarming motility is also observed, where very low levels of DegU∼P support swarming and excessive DegU∼P abolishes swarming. We further demonstrate that the defect of swarming motility in a degU ⁻ strain can be rescued, albeit incompletely, by increased expression of an exogenous epr gene. We also show that an additional extracellular factor(s), apart from epr , regulated by DegU, is required for robust swarming.     

Social interactions in myxobacterial swarming

Swarming, a collective motion of many thousands of cells, produces colonies that rapidly spread over surfaces. In this paper, we introduce a cell-based model to study how interactions between neighboring cells facilitate swarming. We chose to study Myxococcus xanthus, a species of myxobacteria, because it swarms rapidly and has well-defined cell–cell interactions mediated by type IV pili and by slime trails. The aim of this paper is to test whether the cell contact interactions, which are inherent in pili-based S motility and slime-based A motility, are sufficient to explain the observed expansion of wild-type swarms. The simulations yield a constant rate of swarm expansion, which has been observed experimentally. Also, the model is able to quantify the contributions of S motility and A motility to swarming. Some pathogenic bacteria spread over infected tissue by swarming. The model described here may shed some light on their colonization process.     

Genetic population structure of Natterer's bats explained by mating at swarming sites and philopatry

During autumn 'swarming', large numbers of temperate bats chase each other in and around underground sites. Swarming has been proposed to be a mating event, allowing interbreeding between bats from otherwise isolated summer colonies. We studied the population structure of the Natterer's bat (Myotis nattereri), a swarming species in northern England, by sampling bats at seven sites in two swarming areas and at 11 summer colonies. Analysis of molecular variance (amova) and genetic assignment analyses showed that the swarming areas (60 km apart) support significantly different populations. A negative correlation was found between the distance of a summer colony from a swarming area and the assignment of bats to that area. High gene diversity was found at all sites (HE = 0.79) suggesting high gene flow. This was supported by a low FST (0.017) among summer colonies and the absence of isolation by distance or substructure among colonies which visit one swarming area. The FST, although low, was significantly different from zero, which could be explained by a combination of female philopatry and male-mediated gene flow through mating at swarming sites with bats from other colonies. Modelling suggested that if effective size of the summer colonies (Ne) was low to moderate (10-30), all mating must occur at the swarming sites to account for the observed FST. If the Ne was higher (50), in addition to random mating during swarming, there may be nonrandom mating at swarming sites or some within-colony mating. Conservation of swarming sites that support potentially large populations is discussed.