High-density Colonies Research Articles

Culture Wars: Empirically Determining the Best Approach for Plasmid Library Amplification.

DNA libraries are critical components of many biological assays. These libraries are often kept in plasmids that are amplified in E. coli to generate sufficient material for an experiment. Library uniformity is critical for ensuring that every element in the library is tested similarly and is thought to be influenced by the culture approach used during library amplification. We tested five commonly used culturing methods for their ability to uniformly amplify plasmid libraries: liquid, semisolid agar, cell spreader-spread plates with high or low colony density, and bead-spread plates. Each approach was evaluated with two library types: a random 80-mer library, representing high complexity and low coverage of similar sequence lengths, and a human TF ORF library, representing low complexity and high coverage of diverse sequence lengths. We found that no method was better than liquid culture, which produced relatively uniform libraries regardless of library type. However, when libraries were transformed with high coverage, the culturing method had minimal impact on uniformity or amplification bias. Plating libraries was the worst approach by almost every measure for both library types and, counterintuitively, produced the strongest biases against long sequence representation. Semisolid agar amplified most elements of the library uniformly but also included outliers with orders of magnitude higher abundance. For amplifying DNA libraries, liquid culture, the simplest method, appears to be best.

A simple, robust, broadly applicable insertion mutagenesis method to create random fluorescent protein: target protein fusions.

A simple, broadly applicable method was developed using an in vitro transposition reaction followed by transformation into Escherichia coli and screening plates for fluorescent colonies. The transposition reaction catalyzes the random insertion of a fluorescent protein open reading frame into a target gene on a plasmid. The transposition reaction is employed directly in an E. coli transformation with no further procedures. Plating at high colony density yields fluorescent colonies. Plasmids purified from fluorescent colonies contain random, in-frame fusion proteins into the target gene. The plate screen also results in expressed, stable proteins. A large library of chimeric proteins was produced, which was useful for downstream research. The effect of using different fluorescent proteins was investigated as well as the dependence of the linker sequence between the target and fluorescent protein open reading frames. The utility and simplicity of the method were demonstrated by the fact that it has been employed in an undergraduate biology laboratory class without failure over dozens of class sections. This suggests that the method will be useful in high-impact research at small liberal arts colleges with limited resources. However, in-frame fusion proteins were obtained from 8 different targets suggesting that the method is broadly applicable in any research setting.

Quantifying dispersal between two colonies of northern elephant seals across 17 birth cohorts.

Dispersal drives extinction-recolonization dynamics of metapopulations and is necessary for endangered species to recolonize former ranges. Yet few studies quantify dispersal and even fewer examine consistency of dispersal over many years. The northern elephant seal (Mirounga angustirostris) provides an example of the importance of dispersal. It quickly recolonized its full range after near extirpation by 19th century hunting, and though dispersal was observed it was not quantified. Here we enumerate lifetime dispersal events among females marked as pups at two colonies during 1994-2010, then correct for detection biases to estimate bidirectional dispersal rates. An average of 16% of females born at the Piedras Blancas colony dispersed northward 200 km to breed at Año Nuevo, while 8.0% of those born at Año Nuevo dispersed southward to Piedras Blancas. The northward rate fluctuated considerably but was higher than southward in 15 of 17 cohorts. The population at Piedras Blancas expanded 15-fold during the study, while Año Nuevo's declined slightly, but the expectation that seals would emigrate away from high density colonies was not supported. During the 1990s, dispersal was higher away from the small colony toward the large. Moreover, cohorts born later at Piedras Blancas, when the colony had grown, dispersed no more than early cohorts. Consistently high natal dispersal in northern elephant seals means the population must be considered a single large unit in terms of response to environmental change. High dispersal was fortuitous to the past recovery of the species, and continued dispersal means elephant seals will likely expand their range further.

Sibling competition for food and kin selection in nestlings of a colonial raptor

Kin selection theory predicts that individual progeny members, in species where offspring share depreciable parental care, should increasingly attempt to monopolize resources provided by the parents as their relatedness to brood competitors declines. Although the level of relatedness among broodmates is often variable in vertebrates, whether it affects within-brood competition dynamics has seldom been studied. We tested this prediction in nestlings of the lesser kestrel, Falco naumanni , a cavity-nesting colonial raptor that can breed in high-density colonies, where nestlings often move between nearby nests, generating mixed parentage among broodmates. Pairs of nestlings composed of either unrelated, unfamiliar individuals or siblings were experimentally forced to compete for food. Nestlings stole prey more frequently when competing with an unrelated, unfamiliar competitor than when competing with a sibling. This finding adds to the limited evidence that rivalry among broodmates is affected by their relatedness, suggesting that kin recognition and kin selection may play an important role in determining individual competitive strategies and care allocation in multiparous species. • Coloniality favours the occurrence of mixed kinship broods. • Competition for food should differ between siblings and unrelated offspring. • We tested this prediction in mixed kinship lesser kestrel broods. • Prey thefts were more frequent when competitors were unrelated. • Kin selection affects competitive behaviour in altricial avian offspring.

Designing crop pollination services: A spatially explicit agent-based model for real agricultural landscapes

The decline of pollinators is a widespread problem in today's agriculture, affecting the yield of many crops. Improved pollination management is therefore essential, and honey bee colonies are often used to improve pollination levels. In this work, we applied a spatially explicit agent-based model for the simulation of crop pollination by honey bees under different management scenarios and landscape configurations. The model includes 1) a representation of honey bee social dynamics; 2) an explicit representation of resource dynamics; 3) a probabilistic approach to the foraging site search process; and 4) a mechanism of competition for limited resources. We selected 60 sample units from the rural landscape of the Chilean region with the largest apple-growing area and evaluated the effectiveness of different pollination strategies in terms of number of visits and number of pollinated flowers per hectare of apple crops. Finally, we analyzed how the effects of these practices depended on the structure of adjacent landscapes. Higher colony density per hectare in the focal crop increased the number of honey bee visits to apple inflorescences; however, the effects were nonlinear for rates of pollinated flowers, suggesting that there is an optimum beyond which a greater number of honey bees does not signify increased levels of crop pollination. Furthermore, high relative proportions of mass flowering crops and natural habitats in the landscape led to a decrease in honey bee densities in apple fields in landscapes with high relative cover of apple orchards (dilution effect). Our results indicate that for optimal crop pollination, strategies for management of pollinator species should consider the modulating effects of the surrounding landscape on pollination effectiveness. This model could thus be a useful tool to help farmers, beekeepers, and policy-makers plan the provision of pollination services, while also promoting the biodiversity and sustainability of agroecosystems.

Assessing colony elimination in multicolonial ants: Estimating field efficacy of insecticidal baits against the invasive dark rover ant (Brachymyrmex patagonicus).

A frequent goal of pest management strategies targeting social insects is total colony elimination. Insecticidal baits are highly effective at controlling social insect pests, although their ability to provide total colony elimination has only been well studied in a few species. Genetically testing colony elimination in many urban pest ants can be challenging due to indistinct colony boundaries observed in unicolonial, invasive species; however, some pest ants, such as the dark rover ant (Brachymyrmex patagonicus), maintain strict colony borders through aggression towards non-nestmates. Each of these distinct colonies can be identified using molecular markers, allowing for the tracking of individual colonies pre- and post-treatment to measure colony density. While counting the number of foraging workers to assess treatment efficacy may suffice in some cases, it offers little insight into the colony-level impacts of a treatment. Using microsatellite markers, distinct rover ant colonies were identified and tracked around residential structures before and after the application of an imidacloprid bait. The number of foraging ants at the treated structures was reduced by an average of 83.0% over a 28-day observation period. Baiting also significantly reduced the total number of colonies present. At the treatment structures, only ~25% of the original colonies remained at the end of the study. Colonies with foraging trails <1.5m from a bait station had a higher chance of being eliminated. Using insecticidal baits against B. patagonicus can be highly effective at colony elimination; however, with such small foraging ranges and high colony densities, proper placement is required to ensure enough bait is properly positioned to treat all colonies affecting a structure. © 2022 Society of Chemical Industry.

Maternal responses to pup calls in a high-cost lactation species.

Bonding between mothers and their young is fundamental to mammalian reproductive behaviour and individual fitness. In social systems where the risk of confusing filial and non-filial offspring is high, mothers should demonstrate early, strong and consistent responses to their kin throughout the period of offspring dependence, irrespective of maternal traits. We tested this hypothesis through playback experiments in the northern elephant seal Mirounga angustirostris, a phocid species that breeds in high-density colonies. We found that mothers recognized their offspring throughout lactation and as early as 1-2 days after parturition. Measures of experience (age) and temperament (aggressivity) did not predict their response strength to filial playback treatments, nor did pup age or sex. Some mothers showed great consistency in behavioural responses throughout the lactation period, while others were less predictable. The strength of a female's response did not influence her pup's weaning weight; however, more consistent females weaned pups of higher mass. This is a rare demonstration of individual recognition among phocid mothers and their offspring, and suggests that consistency in maternal responsiveness may be an important social factor influencing the pup's growth and survival.

Curing two predominant viruses occurring in Lentinula edodes by chemotherapy and mycelial fragmentation methods

Previous research has established that Lentinula edodes mycovirus HKB (LeV-HKB) and L. edodes partitivirus 1(LePV1) are major mycoviruses identified in L.edodes germplasm. In this paper, two different methods for curing these two dsRNA mycoviruses, ribavirin treatment and mycelial fragmentation, were evaluated for the first time. Mycelial fragmentation was found to resulted in LeV-HKB- and LePV1-cured fungal strains, whereas ribavirin treatment could eliminate LeV-HKB only. Although no LePV1-cured strain was obtained via ribavirin treatment by the end of the experiment, the relative LePV1 concentration in the eighth successive subcultures was lower than that of the untreated control. The culture features of several virus-cured strains had faster mycelial growth rate and higher colony density than the infected ones. It was also suggested that LeV-HKB infection may affect the pigmentation in plate- and bag-cultivated mycelia of L. edodes strain L135.

Honey bee (Apis mellifera) colony strength and its effects on pollination and yield in highbush blueberries (Vaccinium corymbosum).

Many pollination studies with honey bees have examined the effect of colony density on crop yield and yet overlook the effect of variation in the population size of these colonies. High colony density in northern highbush blueberry has been met with concerns from beekeepers who feel higher densities will intensify outbreaks of European foulbrood (EFB, Melissococcus plutonius, Truper and dé Clari), a honey bee brood disease. The purpose of this study was to confirm the prevalence of EFB in colonies pollinating blueberries and to determine whether field-level variation in the population of adult workers in colonies explained variation in blueberry fruit set and/or yield. We addressed these objectives over the course of two production seasons at 13 commercial blueberry fields in Oregon, USA, stocked with identical densities of 10 colonies/ha. We confirmed that all colonies had negligible symptoms of EFB at the start of blueberry pollination, but 53% of colonies in 2019 and 41% in 2020 had symptoms immediately following the pollination season. We also validated a method for rapidly assessing adult honey bee colony populations, namely by counting the rate of foragers returning to colonies, and it was found to be strongly correlated to true internal adult bee population independent of year and ambient temperature at the time of evaluation. Using returning forager counts, we determined there was considerable variation in the average population of colonies at each field, ranging from an estimated 10,300 to 30,700 adult worker bees per colony. While average colony strength did not predict variation in fruit set, it was related to variation in yield, independent of year. Our linear model of flight count (as a proxy for colony strength) predicts estimated yield increases of up to 25,000 kg/ha of blueberries could be achieved by colonies stronger than the recommended six frame minimum, suggesting that higher pollination benefits could be achieved without increasing hive density if stronger colonies are promoted.

Honey bee virus causes context-dependent changes in host social behavior

Anthropogenic changes create evolutionarily novel environments that present opportunities for emerging diseases, potentially changing the balance between host and pathogen. Honey bees provide essential pollination services, but intensification and globalization of honey bee management has coincided with increased pathogen pressure, primarily due to a parasitic mite/virus complex. Here, we investigated how honey bee individual and group phenotypes are altered by a virus of concern, Israeli acute paralysis virus (IAPV). Using automated and manual behavioral monitoring of IAPV-inoculated individuals, we find evidence for pathogen manipulation of worker behavior by IAPV, and reveal that this effect depends on social context; that is, within versus between colony interactions. Experimental inoculation reduced social contacts between honey bee colony members, suggesting an adaptive host social immune response to diminish transmission. Parallel analyses with double-stranded RNA (dsRNA)-immunostimulated bees revealed these behaviors are part of a generalized social immune defensive response. Conversely, inoculated bees presented to groups of bees from other colonies experienced reduced aggression compared with dsRNA-immunostimulated bees, facilitating entry into susceptible colonies. This reduction was associated with a shift in cuticular hydrocarbons, the chemical signatures used by bees to discriminate colony members from intruders. These responses were specific to IAPV infection, suggestive of pathogen manipulation of the host. Emerging bee pathogens may thus shape host phenotypes to increase transmission, a strategy especially well-suited to the unnaturally high colony densities of modern apiculture. These findings demonstrate how anthropogenic changes could affect arms races between human-managed hosts and their pathogens to potentially affect global food security.

The rise of octocoral forests on Caribbean reefs.

Coral reefs throughout the tropics have experienced large declines in the abundance of scleractinian corals over the last few decades, and some reefs are becoming functionally dominated by animal taxa other than scleractinians. This phenomenon is striking on many shallow reefs in the tropical western Atlantic, where arborescent octocorals now are numerically and functionally dominant. Octocorals are one of several taxa that have been overlooked for decades in analyses of coral reef community dynamics, and our understanding of why octocorals are favoured (whereas scleractinians are not) on some modern reefs, and how they will affect the function of future reef communities, is not commensurate with the task of scientifically responding to the coral reef crisis. We summarize the biological and ecological features predisposing octocorals for success under contemporary conditions, and focus on those features that could have generated resistance and resilience of octocoral populations to environmental change on modern reefs. There is a rich set of opportunities for rapid advancement in understanding the factors driving the success of octocorals on modern reefs, but we underscore three lines of inquiry: (1) the functional implications of strongly mixotrophic, polytrophic, and plastic nutrition, (2) the capacity to recruit at high densities and maintain rapid initial rates of vertical growth, and (3) the emergent properties associated with dense animal forests at high colony densities.

High-resolution maps of Swiss apiaries and their applicability to study spatial distribution of bacterial honey bee brood diseases.

Honey bees directly affect and are influenced by their local environment, in terms of food sources, pollinator densities, pathogen and toxin exposure and climate. Currently, there is a lack of studies analyzing these data with Geographic Information Systems (GIS) to investigate spatial relationships with the environment. Particularly for inter-colonial pathogen transmission, it is known that the likelihood of a healthy colony to become infested (e.g., Varroosis) or infected (e.g., American foulbrood—AFB, European foulbrood—EFB) increases with higher colony density. Whether these transmission paths can actually be asserted at apiary level is largely unknown. Here, we unraveled spatial distribution and high-resolution density of apiaries and bacterial honey bee brood diseases in Switzerland based on available GIS data. Switzerland as ‘model country’ offers the unique opportunity to get apiary data since 2010 owing to compulsory registration for every beekeeper. Further, both destructive bee brood diseases (AFB and EFB) are legally notifiable in Switzerland, and EFB has an epizootic character for the last decades. As governmental data sets have to be ameliorated, raw data from the cantonal agricultural or veterinary offices have been included. We found a mean density of 0.56 apiaries per km2, and high resolution spatial analyzes showed strong correlation between density of apiaries and human population density as well as agricultural landscape type. Concerning two bacterial bee brood diseases (AFB, EFB), no significant correlation was detectable with density of apiaries on cantonal level, though a high correlation of EFB cases and apiary density became obvious on higher resolution (district level). Hence, Swiss EFB epizootics seem to have benefited from high apiary densities, promoting the transmission of pathogens by adult bees. The GIS-based method presented here, might also be useful for other bee diseases, anthropogenic or environmental factors affecting bee colonies.

Revisiting the Role of Aquatic Plants in Beaver Habitat Selection

Beaver (Castor canadensis) can help to restore wetlands and mitigate the effects of climate change on hydrological regimes in some areas. Therefore, ongoing resource management efforts seek to promote the presence and persistence of beaver populations. The long-term success of such endeavors requires an understanding of what conditions are conducive to sustaining beaver populations, often in landscapes with degraded forest resources. Available information about beaver habitat suitability is largely based on short-term studies in stream habitats that do not consider aquatic plant suitability, even though aquatic plants may comprise over half of beavers' annual diets. In the present study, we assess whether the availability of woody plants, conditions conducive to the availability of aquatic vascular plants (macrophytes), and/or other features of basin morphology are associated with the persistence and density of beaver occupancy in 23 lakes over a 50-y period. We incorporate field-based study that includes all species of macrophytes, to extend previous work that focused specifically on water lily species (Nymphaea spp., Nuphar spp.). Lake perimeter (a function of sinuosity and surface area) and total macrophyte cover were associated with the persistence and density of beaver occupancy in lakes. Each factor independently explained ∼70% (R2adj) of the variation in the persistence of beaver occupancy. Percent cover of floating-leaved macrophytes was a leading predictor of beaver colony density in lakes, independently explaining 72% (R2adj) of the variation. Brasenia schreberi appeared particularly valuable to sustaining beaver in smaller lakes. Several lakes with abundant Brasenia supported high colony densities and long-term colony occupancy. Where feasible, beaver restoration efforts may increase the probability of success by facilitating beaver access to lakes that host key rhizomatous macrophyte species.

Real-time PCR as a decision aid in the control of European foulbrood

The number of cases of European foulbrood (EFB) increased massively in Switzerland since 2000. According to Swiss legislation, the health status of all colonies within one kilometer from a case needs to be identified by a bee inspector. This results in a workload too high to be handled in due time in areas with a high prevalence and high colony density. Therefore, the primary goal of this study was testing the hypothesis that the number of visual inspections could be reduced if brood nest bees sampled by owners were tested by real-time PCR for the presence of Melissococcus plutonius, and results were assigned to one of the three categories “negative”, “positive,” or “strong positive” depending on the amount of bacterial DNA. Only apiaries with strong positive results were inspected visually. Furthermore, the development of the bacterial load over time was analyzed on an apiary level to determine whether this would allow predicting the outcome of an infection. For this purpose, bee samples of 88 apiaries were assayed by real-time PCR and visually inspected in parallel over several months. Comparison of the two methods revealed a sensitivity of the real-time PCR of 93.5% and a specificity of 58.6% relative to visual inspection. One month after first sampling the bacterial load was still on the same level in 54%, had increased in 17%, and decreased in 29% of 35 apiaries. Establishing predictions on the basis of real-time PCR results is therefore not possible. In conclusion, due to its poor specificity, preliminary real-time PCR would not allow reducing the number of visual inspections sufficiently to compensate the costs of additional laboratory diagnosis. However, real-time PCR may be a valuable tool for identifying infected colonies in the context of trade and migratory beekeeping.

First- and second-order sociality determine survival and reproduction in cooperative cichlids.

Cooperative breeders serve as a model to study the evolution of cooperation, where costs and benefits of helping are typically scrutinized at the level of group membership. However, cooperation is often observed in multi-level social organizations involving interactions among individuals at various levels. Here, we argue that a full understanding of the adaptive value of cooperation and the evolution of complex social organization requires identifying the effect of different levels of social organization on direct and indirect fitness components. Our long-term field data show that in the cooperatively breeding, colonial cichlid fish Neolamprologus pulcher, both large group size and high colony density significantly raised group persistence. Neither group size nor density affected survival at the individual level, but they had interactive effects on reproductive output; large group size raised productivity when local population density was low, whereas in contrast, small groups were more productive at high densities. Fitness estimates of individually marked fish revealed indirect fitness benefits associated with staying in large groups. Inclusive fitness, however, was not significantly affected by group size, because the direct fitness component was not increased in larger groups. Together, our findings highlight that the reproductive output of groups may be affected in opposite directions by different levels of sociality, and that complex forms of sociality and costly cooperation may evolve in the absence of large indirect fitness benefits and the influence of kin selection.

Generating size-controlled embryoid bodies using laser direct-write

Embryonic stem cells (ESCs) have the potential to self-renew and differentiate into any specialized cell type. One common method to differentiate ESCs in vitro is through embryoid bodies (EBs), three-dimensional cellular aggregates that spontaneously self-assemble and generally express markers for the three germ layers, endoderm, ectoderm, and mesoderm. It has been previously shown that both EB size and 2D colony size each influence differentiation. We hypothesized that we could control the size of the EB formed by mouse ESCs (mESCs) by using a cell printing method, laser direct-write (LDW), to control both the size of the initial printed colony and the local cell density in printed colonies. After printing mESCs at various printed colony sizes and printing densities, two-way ANOVAs indicated that the EB diameter was influenced by printing density after three days (p = 0.0002), while there was no effect of the printed colony diameter on the EB diameter at the same timepoint (p = 0.74). There was no significant interaction between these two factors. Tukey's honestly significant difference test showed that high-density colonies formed significantly larger EBs, suggesting that printed mESCs quickly aggregate with nearby cells. Thus, EBs can be engineered to a desired size by controlling printing density, which will influence the design of future differentiation studies. Herein, we highlight the capacity of LDW to control the local cell density and colony size independently, at prescribed spatial locations, potentially leading to better stem cell maintenance and directed differentiation.

Parasite communities in stray cat populations from Lisbon, Portugal

Stray cats live in high-density colonies in urban areas and pose a health hazard to household cats and humans. In Portugal, information on the parasitic fauna of stray cats is limited and relies mostly on results from faecal analysis. The present survey aimed to determine the prevalence, diversity and intensity of parasites in stray cats from the urban area of Lisbon by means of parasitological necropsy. Internal organs were collected from 162 cats captured in different areas of the city and systematically subjected to parasitological dissection. Helminths were identified by macro- and microscopic examination and protozoa by faecal floatation and sedimentation techniques. The overall prevalence of parasites was 90.7% (95% confidence interval (CI): 85.3-94.6%). A total of 12 parasite species was recorded: Cystoisospora felis (14.2%), Cystoisospora rivolta (46.3%), Sarcocystis sp. (1.2%), Ancylostoma tubaeforme (19.1%), Toxocara cati (38.3%), Ollulanus tricuspis (30.9%), Aelurostrongylus abstrusus (12.4%), Eucoleus aerophilus (0.6%), Taenia taeniaeformis (3.1%), Dipylidium caninum (53.1%), Joyeuxiella pasqualei (15.4%) and Diplopylidium nölleri (3.7%). Overall mean species richness was 2.36 ± 1.52. Helminth mean intensity was highest for O. tricuspis (285.8), followed by D. caninum (42.4), J. pasqualei (14.4), A. tubaeforme (8.1) and T. cati (5.9). The prevalence and variety of parasites found in our sampling are substantially higher than the numbers previously reported in Portugal. Some of the parasites, including T. cati and A. tubaeforme, are zoonotic, which emphasizes the need for parasite control strategies based on demographic containment of stray cat populations in urban areas to promote public health protection.

Nesting Associations without Interdependence: A Preliminary Review on Plesiobiosis in Ants

Plesiobiosis, the most basic form of interspecific associations in ants, denotes occasional or regular nesting of heterospecific colonies of certain species pairs in close proximity to each other without biological interdependence. Plesiobionts differ from each other both in morphology and in behaviour (e.g., in their foraging strategies), and at least one of the plesiobiotic pair is a submissive species. Recent studies on plesiobiosis have revealed thatFormica fuscaandLasius flavusare two of the most frequent plesiobionts. To date, at least 48 different plesiobiotic species pairs have been recorded from various habitat types of the Holarctic region. Two main habitat properties may play a role in the forming of plesiobiosis: the scarcity of suitable nesting sites as a forcing factor and the sufficient amount of food sources available, influencing the abundance of colonies. Thus, high colony density may contribute to the formation of such associations, resulting in (1) frequent nesting in each other's neighbourhood and (2) stronger intraspecific competition, which forces colonies into the vicinity of heterospecific nests. Plesiobiotic associations formed this way may promote persistent coexistence, leading to the formation of other types of interspecific associations (e.g., clepto- or lestobiosis).

Estimating density of a forest‐dwelling bat: a predictive model for Rafinesque's big‐eared bat

Data on species distribution and abundance are the foundation of population ecology. However, due to difficulties in surveying bats, abundance estimates for tree-roosting microchiropterans are non-existent. Therefore, our objective was to develop methods for estimating colony abundance and density, taking as our model Rafinesque's big-eared bat Corynorhinus rafinesquii, a species of conservation concern found in cypress-gum swamps of the southeastern United States. We searched 123 transects at eight study sites in the Coastal Plain of Georgia, USA to locate and characterize diurnal summer roosts of C. rafinesquii. We modeled the relationship between the number of bat colonies and landscape-scale habitat variables with zero-inflated negative binomial regression and used Akaike's information criterion to select the most parsimonious models. We generated a predictive density map to identify areas of high colony density and to estimate overall abundance. Colony density was predicted by the duration of wetland flooding, wetland width, and study site. Application of the regression model to a GIS indicated there were 3,734 colonies containing 6,910 adult bats on the eight study sites. Predicted density ranged from 0.07 colonies/ha and 0.07 adult bats/ha in saturated wetlands to 0.47 colonies/ha and 1.18 adult bats/ha in semi-permanently flooded wetlands. This study is the first to estimate density and abundance of forest-dwelling microchiropterans over a large area. Such data can serve as a baseline for future work on population trends in C. rafinesquii. In addition, our approach could be replicated for other bat species with moderately cryptic roosts.

Spatial-temporal dynamics of collective chemosensing

Although the process of chemosensing by individual cells is intrisically stochastic, multicellular organisms exhibit highly regulated responses to external stimulations. Two key elements to understand the deterministic features of chemosensing are intercellular communications and the role of pacemaker cells. To characterize the collective behavior induced by these two factors, we study the spatial-temporal calcium dynamics of fibroblast cells in response to ATP stimulation. We find that closely packed cell colonies exhibit faster, more synchronized, and highly correlated responses compared to isolated cells. In addition, we demonstrate for chemosensing the existence of pacemaker cells and how the presence of gap junctions impact the first step of the collective response. By further comparing these results with the calcium dynamics of cells embedded in thin hydrogel films, where intercellular communication is only possible via diffusing molecules, we conclude that gap junctions are required for synchronized and highly correlated responses among cells in high density colonies. In addition, in high density cell colonies, both communication channels lead to calcium oscillations following the stimulation by external ATP. While the calcium oscillations associated with cells directly exposed to external flows were transient, the oscillations of hydrogel trapped cells can persist with a fundamental frequency and higher harmonics. Our observations and measurements highlight the crucial role of intercellular signaling for generating regulated spatial and temporal dynamics in cell colonies and tissues.