Testing for context-dependence in a processing chain interaction among detritus-feeding aquatic insects

Abstract 1. Scirtid beetles may benefit mosquitoes Ochlerotatus triseriatus (Say) by consuming whole leaves and leaving behind fine particles required by mosquito larvae. Such interactions based on the sequential use of a resource that occurs in multiple forms are known as processing chains.
2. Models of processing chains predict that interactions can vary from commensal (0, +) to amensal (0, –), depending on how quickly resource is processed in the absence of consumers.
3. The scirtid– O. triseriatus system was used to test the prediction derived from processing chain models that, as consumer-independent processing increases, scirtids benefit mosquitoes less. Consumer-independent processing rate was manipulated by using different leaf species that vary in decay rate, or by physically crushing a single leaf type to different degrees.
4. Although scirtids increased the production of fine particles, the effects of scirtids on mosquitoes were weak and were not dependent on consumer-independent processing rate.
5. In the leaf manipulation experiment, a correlation between scirtid feeding and consumer-independent processing was detected. Numerical simulations suggest that such a correlation may eliminate shifts from commensal to amensal at equilibrium; because mosquito populations are typically not at equilibrium, however, this correlation may not be important.
6. There was evidence that mosquitoes affected scirtids negatively, which is inconsistent with the structure of processing chain interactions in models. Processing chain models need to incorporate more detail on the biology of scirtids and O. triseriatus , especially alternative mechanisms of interaction, if they are to describe scirtid– O. triseriatus dynamics accurately.     

Interactive effects of pH and leaf litter on a shredder, the scirtid beetle, Helodes pulchella, inhabiting tree-holes

1. Scirtid beetles (Coleoptera: Scirtidae) are common inhabitants of water-filled treeholes and interact with other detritivores in a processing chain commensalism. The strength of the commensalism is determined by resource quantity and the organisms involved have different tolerances to low pH. To determine the effects and interactions of resource quantity and pH on one of these leaf-shredding scirtid beetles, Helodes pulchella (Guerin), leaf litter and pH were varied independently in experimental microcosms which mimic treehole habitats.
2. Helodes pulchella growth was affected by both resources and pH in two similar experiments conducted in different years. Scirtid larvae grew more slowly in low resource treatments and also grew more slowly in low pH treatments. Scirtid survival was not affected by pH, but was lower in high resource microcosms in the 1997 experiment. Consumption of leaf litter was much higher in high resource microcosms, even though not all leaf litter was consumed in low leaf litter treatments.
3. Thus, water chemistry and leaf litter quantity could have strong effects on resource availability through effects on these beetle facilitators, in turn affecting other insects living in these detritus-based communities.     

Natural and manipulated populations of the treehole mosquito, Ochlerotatus triseriatus, at its northernmost range limit in southern Ontario, Canada.

Ochlerotatus triseriatus, the eastern treehole mosquito, reaches its northernmost range limit in the extreme southeast of Canada. As a known vector of West Nile and La Crosse encephalitis viruses and a potential vector of eastern equine encephalitis, its population biology is of interest. In southern Ontario, high larval densities occur in urban woodlots within sugar maple and American beech treehole communities comprising rotifers, nematode worms, mites, other dipterans, and scirtid beetles. Treehole water was characterized by low dissolved oxygen levels and seasonally variable pH and temperature, with the latter being most influential on local populations. Densities were significantly higher (up to 503 larvae 100 ml(-1)) in tree holes close to the forest floor (<1 m) and in experimental tree holes seeded with autumn-shed maple leaves as opposed to leaves of black oak and beech. In this locality, weekly sampling showed Oc. triseriatus to be multivoltine, with mass egg hatching beginning under coldwater (<10 degrees C) conditions in March/April, and thereafter producing three successful generations with a possible, less successful fourth in late summer. Some 1st instar larvae were present in water as cold as 0.7 degree C. Compared with larval psychodids living in the same tree hole, population losses of Oc. triseriatus due to washout during major rainfall events were negligible despite high flowthrough of water derived from stemflow.     

Interactions of Salmonella enterica with lettuce leaves

Aims:  To investigate the interactions of Salmonella enterica with abiotic and plant surfaces and their effect on the tolerance of the pathogen to various stressors.
Methods and Results:  Salmonella strains were tested for their ability to form biofilm in various growth media using a polystyrene plate model. Strong biofilm producers were found to attach better to intact Romaine lettuce leaf tissue compared to weak producers. Confocal microscopy and viable count studies revealed preferential attachment of Salmonella to cut-regions of the leaf after 2 h at 25°C, but not for 18 h at 4°C. Storage of intact lettuce pieces contaminated with Salmonella for 9 days at 4°C resulted only in small changes in population size. Exposure of lettuce-associated Salmonella cells to acidic conditions (pH 3·0) revealed increased tolerance of the attached vs planktonic bacteria.
Conclusions:  Biofilm formation on polystyrene may provide a suitable model to predict the initial interaction of Salmonella with cut Romaine lettuce leaves. Association of the pathogen with lettuce leaves facilitates its persistence during storage and enhances its acid tolerance.
Significance and Impact of the Study:  Understanding the interactions between foodborne pathogens and lettuce might be useful in developing new approaches to prevent fresh produce-associated outbreaks.     

Inclusion of biotic stress (consumer pressure) alters predictions from the stress gradient hypothesis

1.  The stress gradient hypothesis (SGH) predicts a shift from net negative interactions in benign environments towards net positive in harsh environments in ecological communities. While several studies found support for the SGH, others found evidence against it, leading to a debate on how nature and strength of species interactions change along stress gradients, and to calls for new empirical and theoretical work.
2.  In the latest attempt in this journal, it is successfully argued how the SGH should be expanded by considering different life strategies of species (stress tolerance versus competitive ability) and characteristics of abiotic stress (resource versus non-resource based) over wider stress gradients (opposed to low–high contrasts), but the crucial role of biotic stress by consumers is largely ignored in this refinement.
3.  We point out that consumers strongly alter the outcome of species interactions in benign and harsh environments, and show how inclusion of consumer-incurred biotic stress alters the predicted outcome of interactions along resource- and non-resource-based stress gradients for stress-tolerant and competitive benefactors and beneficiaries.
4.   Synthesis. New studies should include stress gradients consisting of both abiotic and biotic components to disentangle their impacts, and to improve our understanding of how species interactions change along environmental gradients.     

Anti-Helicobacter pylori activity of Lactobacillus delbrueckii subsp. bulgaricus strains: preliminary report

Aims:  To evaluate the activities of six Lactobacillus delbrueckii subsp. bulgaricus (LB) strains against 30 Helicobacter pylori strains by agar-well diffusion method.
Methods and Results:  LB cultures [4 × 10⁸–4 × 10⁹ CFU ml⁻¹) either were prepared in milk at their native pH, 3·8–5·0, or were adjusted to pH 6·4–7·7. At low and neutralized pH, LB strains inhibited the growth by 40–86·7% and 16·7–66·7% of H. pylori strains, respectively. LB activity was strain-dependent. At low and neutralized pH, one and five H. pylori strains, respectively, were not inhibited by any LB strain. LB2 and LB3, taken together, were active against most metronidazole and clarithromycin resistant strains.
Conclusions:  All LB strains inhibited a number of H. pylori strains, including also antibiotic resistant strains. LB activity was strain-dependent and better at low pH. At low pH values, the most active LB strains were LB1, LB2 and LB3, inhibiting 86·7% of H. pylori strains, while at neutralized pH values, the most active LB strains were LB2 and LB3, inhibiting 53·3 and 66·7% of H. pylori strains, respectively.
Significance and Impact of the Study:  LB could be utilized in the treatment or prophylaxis of H. pylori infection and warrants clinical investigations.     

Characterization of some culture factors affecting oxalate degradation by the mycoparasite Coniothyrium minitans

Aims:  To find possible approaches to utilize the mechanism of oxalate degradation by Coniothyrium minitans (Cm) in controlling the plant pathogen Sclerotinia sclerotiorum (Ss).
Methods and Results:  Differences in oxalate degradation by different Cm strains and effects of the initial oxalate concentration, ambient pH and nutrient factors on mycelial growth and oxalate degradation by Cm were studied in shaken cultures. Results showed that two wild-type Cm strains, Chy-1 and ZS-1, did not differ in oxalate degradation in modified potato dextrose broth (mPDB) amended with oxalic acid (OA). Cm could grow in mPDB amended with sodium oxalate (SO-mPDB) at pH 6·5 or with ammonium oxalate (AO-PDB) at pH 6·2, but oxalate degradation was very low; oxalate degradation was greatly enhanced in SO- or AO-mPDB with pH being lowered to 2·8–2·9. Similarly, oxalate degradation was higher than 90% in OA-amended mPDB at pH 4·4 but was reduced to be <22% at pH 7·0. Five carbon sources and three nitrogen sources investigated and nutrients from mycelia and sclerotia of Ss were favorable for the growth of Cm and OA degradation by Cm.
Conclusions:  Cm can degrade oxalate under acidic pH. Exudates from mycelia or sclerotia of Ss may serve as nutrients for Cm mycelial growth and degradation of oxalate secreted by Ss.
Significance and Impact of the Study:  The finding of oxalate degradation laid a foundation for mining-related genes in Cm for engineering plant resistance against Ss. Elucidation of the importance of acidic pH and nutrients from Ss in oxalate degradation by Cm will help to understand the interaction between Cm and Ss.     

Variation in the effects of vegetation and litter on recruitment across productivity gradients

1 We tested predictions about how the effect of vegetation and litter on seedling establishment varies among sites and herbaceous community types (sand barrens, prairies, fens). For both vegetation and litter, we also separated direct interactions from indirect interactions and interaction modifications along the gradient.
2 Although the intensity of the effects varied across sites, the direct effects of vegetation or litter alone were consistently facilitative along the productivity gradient. Predominance of facilitative effects may be due to the focus on the seedling establishment phase.
3 However, inclusion of indirect interactions and interaction modifications caused the net effects of both vegetation and litter to become largely negative. While one layer of biomass may be advantageous to ameliorate some moisture stress, the addition of another layer may be disadvantageous if this layer limits light proportionally more than it relieves moisture stress.
4 One exception to this pattern occurred at high productivity when the net effect of vegetation, even in the presence of litter, remained facilitative. The net effect of vegetation was competitive at low productivity and grew increasingly facilitative with productivity. Thus, indirect effects of litter may alter interaction patterns across this gradient.     

Direct and indirect effects of soil structure on the density of an antlion larva in a tropical dry forest

Abstract.  1. We used structural equation models to discriminate direct and indirect effects of soil structure on the abundance of the antlion Myrmeleon crudelis , a neuropteran larva that digs conical pits in soil to capture small arthropods. We proposed that soil structure may modify antlion density indirectly through its influence on tree cover, which in turn directly alters the amount of sun and rain that can reach the forest floor and the amount of litter fall.
2. The proportion of finer soils positively affected antlion density directly, but negatively tree cover. Tree cover positively affected both the amount of leaf litter and antlion density. Leaf litter negatively affected antlion density. The indirect effects of soils varied in strength and sign depending on whether trees are considered shelters against sun and rain, or leaf litter sources. The relative importance of these effects might also vary between years and seasons.
3. Antlions may select patches of finer soils not only because they are easy substrates in which to build pits, but also for their indirect benefit as sites with low leaf litter, illustrating how indirect interactions may affect the local abundance of semi-sedentary insects.     

Behavioural responses of larval container mosquitoes to a size-selective predator

Abstract.  1. The hypothesis that size-selective predation and species-specific prey behaviours facilitate the coexistence between larvae of invasive Aedes albopictus (Skuse) and U.S.A.-native Ochlerotatus triseriatus (Say) was tested experimentally with the predator Corethrella appendiculata (Grabham).
2. Larval behaviours associated with a higher risk of predation were identified, and prey behavioural responses were tested in either the physical presence of predators or in water containing predation cues. Larvae that thrashed on container bottoms had a higher risk of being captured by fourth instar C. appendiculata than did larvae resting on the water surface. Ochlerotatus triseriatus , but not A. albopictus , adopted low-risk behaviours in response to water-borne cues to predation. Both prey species reduced risky behaviours in the physical presence of the predator, but O. triseriatus showed a stronger response.
3. The vulnerability of 2nd and 3rd instar prey to predation was compared, and behavioural responses were correlated with prey vulnerability. Second instars of both species were more vulnerable to predation by C. appendiculata than were 3rd instars, and the 3rd instar A. albopictus was more vulnerable than O. triseriatus of the same stage. All instars of O. triseriatus showed a similar reduction of risky behaviours in response to the presence of C. appendiculata despite 4th instar prey being relatively invulnerable to size-selective predation.
4. Weaker predator avoidance, coupled with superior competitive ability, of invasive A. albopictus is likely to contribute to its coexistence with O. triseriatus in containers of the south-eastern U.S.A., where C. appendiculata can be abundant.     

Leaf litter species identity alters the structure of pond communities

The input of leaf litter resources is a major driver of ecosystem processes in terrestrial and freshwater habitats. Although variation exists in the quantity and composition of litter inputs due to natural and anthropogenic causes, few studies have examined how such variation influences the structure and composition of aquatic food webs. Using outdoor mesocosms, we examined the bottom–up effects of 10 chemically distinct tree litter species on microbial, algal, invertebrate and vertebrate fauna found in temperate ponds. We hypothesized that individual litter species, which differ in their traits, would differentially and predictably affect abiotic and biotic elements of pond communities. We further hypothesized that the presence of leaf litter, regardless of species, would elevate resource supply and increase the biomass of community members. Finally, we hypothesized that a mixture of litter species would have non‐additive effects on community responses. We followed the system for > 4 months and measured > 30 abiotic and biotic responses related to primary and secondary production. The different species of leaf litter had major effects on abiotic and biotic responses, including phytoplankton, periphyton, zooplankton, snails, amphipods and tadpoles. Most biological responses were negatively associated with soluble carbon content of litter, or litter decay rate. Other litter traits, including phenolic concentrations and litter C:N were of secondary importance but did exhibit both positive and negative associations with several responses. The absence of litter had pervasive effects on abiotic attributes, but did not promote substantial changes in organism biomass. Most responses to the litter mixture were additive. Our results suggest that changes in temperate forest composition can strongly affect pond communities.     

Methods for the recovery of a model virus from healthcare personal protective equipment

Aims:  To develop methods for recovering a model virus (bacteriophage MS2) from healthcare personal protective equipment (PPE).
Methods and Results:  Nine eluents were evaluated for recovery of infectious MS2 from PPE: 1·5% beef extract (BE) pH 7·5 with and without 0·1% Tween 80, 1·5% BE pH 9·0 with and without 0·1% Tween 80, 3% BE pH 7·5 with and without 0·1% Tween 80, 3% BE pH 9·0 with and without 0·1% Tween 80 and PBS with 0·1% Tween 80. Methods were applied to experimentally contaminated PPE. Elution followed by two-step enrichment assay could recover virus inputs as low as 1·5 log₁₀, and could recover >90% of inoculated virus from used items of experimentally contaminated PPE worn by human volunteers.
Conclusions:  BE was effective for recovering infectious viruses from a range of PPE materials.
Significance and Impact of the Study:  PPE plays a crucial role in interrupting transmission of infectious agents from patients to healthcare workers (HCWs). The fate of micro-organisms when PPE is removed and disposed of has important consequences for infection control. Methods described here can be used to conduct rigorous studies of viral survival and transfer on PPE for risk assessments in infection control and HCW protection.     

Stage-dependent predation on competitors: consequences for the outcome of a mosquito invasion

1. Predator-mediated coexistence occurs when predation allows competitors to coexist, due to preferential consumption of a superior competitor relative to an inferior competitor. Differences between the native treehole mosquito ( Aedes triseriatus ) and the co-occurring Asian tiger mosquito ( Aedes albopictus ) in anti-predatory larval behaviours account, in part, for the greater vulnerability of this invasive species to native predatory midge ( Corethrella appendiculata ). We test the hypothesis that stage-dependent differences in the sizes of A. albopictus and A. triseriatus larvae, relative to the size-limited C. appendiculata , contribute to differential consumption and the likelihood of predator-mediated coexistence of these competitors.
2. In all instars, larvae of A. triseriatus were larger than A. albopictus of the same stage. Third and fourth instar C. appendiculata selectively consumed late-stage A. albopictus in preference to same-stage A. triseriatus . Small, early-stage prey larvae did not differ in vulnerability to predation, but large, late-stage larvae differed significantly in vulnerability to predation, probably owing to size-limited predation by fourth instar C. appendiculata. This effect was less pronounced for third instar C. appendiculata .
3. Prey size, in conjunction with anti-predatory behavioural responses, alters the probability of predator-mediated coexistence. A stage-structured predation model showed that equally vulnerable early stages reduce the range of environmental conditions (productivities) in which predator-mediated coexistence is possible, increasing the likelihood of both competitive exclusion of the resident species or failure of the invasive to establish. These results underscore the importance of stage-dependent interspecific differences in predator–prey interactions for determining how predators may affect community composition.     

Effects of fathead minnows and restoration on prairie wetland ecosystems

SUMMARY 1. Research has shown that fish influence the structure and processes of aquatic ecosystems, but replicated studies at the ecosystem level are rare as are those involving wetlands. Some wetlands of the Prairie Pothole Region (PPR) of North America support fish communities dominated by fathead minnows ( Pimephales promelas ) while others are fishless, providing an opportunity to assess the influence of these fish on wetland ecosystems. Additionally, many wetlands have previously been drained and subsequently restored, but the success of these efforts is poorly known and restoration may be impeded by the presence of fish.
2. We assessed the effects of fathead minnows and drainage by studying 20 semipermanent, prairie wetlands in Minnesota from 1996 to 1999. We used a 2 × 2 factorial design to examine the effects of presence and absence of minnows and drainage history (restored/never drained) on the abundance of aquatic invertebrates and amphibians, as well as on the concentrations of chlorophyll a , total phosphorus, total nitrogen and turbidity in the water column.
3. Results showed that fathead minnows are an important determinant of many biotic and abiotic characteristics of wetlands in the eastern PPR. Wetlands with fathead minnows had fewer aquatic insects, large- and small-bodied cladocerans, calanoid copepods, ostracods and larval tiger salamanders, as well as a higher abundance of corixids and greater turbidity and chlorophyll a . A higher concentration of phosphorus in restored basins was the only consistent effect of past management.
4. Fathead minnows usually dominate fish communities in eastern PPR wetlands where fish are present, and can have several strong ecosystem effects. While abiotic variables are important determinants of ecosystem structure in prairie wetlands, they can be strongly influenced by biotic factors.     

Isolation and partial characterization of novel genes encoding acidic cellulases from metagenomes of buffalo rumens

Aims:  To clone and characterize genes encoding novel cellulases from metagenomes of buffalo rumens.
Methods and Results:  A ruminal metagenomic library was constructed and functionally screened for cellulase activities and 61 independent clones expressing cellulase activities were isolated. Subcloning and sequencing of 13 positive clones expressing endoglucanase and MUCase activities identified 14 cellulase genes. Two clones carried two gene clusters that may be involved in the degradation of polysaccharide nutrients. Thirteen recombinant cellulases were partially characterized. They showed diverse optimal pH from 4 to 7. Seven cellulases were most active under acidic conditions with optimal pH of 5·5 or lower. Furthermore, one novel cellulase gene, C67-1, was overexpressed in Escherichia coli , and the purified recombinant enzyme showed optimal activity at pH 4·5 and stability in a broad pH range from pH 3·5 to 10·5. Its enzyme activity was stimulated by dl -dithiothreitol.
Conclusions:  The cellulases cloned in this work may play important roles in the degradation of celluloses in the variable and low pH environment in buffalo rumen.
Significance and Impact of the Study:  This study provided evidence for the diversity and function of cellulases in the rumen. The cloned cellulases may at one point of time offer potential industrial applications.     

Aerial distribution, flight behaviour and eggload: their inter-relationship during dispersal by the sweetpotato whitefly

1. The aerial distribution of Bemisia tabaci Gennadius (the sweetpotato whitefly) was studied during the early ascent phase of flight, to test the degree to which dispersal patterns reflect the flight behaviour of individuals.
2. Marked whiteflies were trapped at four heights between 0 and 7·2 m above fallow ground, and at six distances between 0 and 100 m from the insect source. Insects were trapped during a 2–3 h period after the initiation of flight activity during the summers of 1995 and 1996.
3. Analysis of trap catch data revealed a clear negative exponential relationship between height and aerial distribution, and a slightly weaker negative power relationship between distance and aerial distribution. Marked insects were caught in the uppermost traps adjacent to the source, indicating that a portion of the population had a strong capacity for ascent out of the flight boundary layer.
4. Eggload decreased with the height, but not the distance, at which whiteflies were trapped. Mean eggload close to the ground was significantly greater than that for those trapped at 4·8 and 7·2 m, supporting the hypothesis that there is a trade-off between flight and oogenesis in weak-flying insects.
5. Air temperatures during the trapping periods were positively correlated with the proportion of male and female B. tabaci caught in the highest traps, but not in the most distant traps.
6. The significance of these results for accurate prediction of whitefly dispersal is discussed, and the importance of individual's behaviour in determining dispersal patterns of small insects is emphasized.     

Growth and reducing capacity of Listeria monocytogenes under different initial redox potential

Aims:  To determine the reducing capacity of Listeria monocytogenes and to highlight the effect of redox potential on its growth parameters.
Methods and Results:  The reducing capacity of L. monocytogenes was monitored in Brain Heart Infusion Broth media at different initial redox potential (Eh) and pH at 37°C. The effect of Eh obtained by gas flushing (air, N₂ and N₂-H₂) or by adding potassium ferricyanide and dithiotreitol in concentration from 1 to 10 mmol l⁻¹on L. monocytogenes growth parameters at pH 6·0, 7·0 and 8·0 was investigated. A total change of 539 mV (±44 mV) from an initial redox value of +330 ± 8 mV to a more negative potential in redox curves was observed resulting from L. monocytogenes growth at pH 7·0 at 37°C. A significant influence of pH and redox potential on L. monocytogenes lag phase of growth was shown ( P  < 0·05).
Conclusions:  Listeria monocytogenes exhibited longer lag phase in reducing conditions and at pH 6·0. The method used to modify the redox potential was shown to have no effect on growth parameters at pH 7·0.
Significance and Impact of the Study:  The provided information on the extending lag time and the possible delayed growth of this major pathogen in reducing conditions might be useful for its control in foods.     

Optimization of a new heteropolysaccharide production by a native isolate of Leuconostoc sp. CFR-2181

Aim:  This work is aimed at optimizing the production of a new heteropolysaccharide (HePS) of Leuconostoc sp. CFR-2181 by standardizing the fermentation conditions in a low cost semi-synthetic medium.
Methods and Results:  Both nutritional and cultural parameters, such as carbon source and its concentration, initial pH of the exopolysaccharide (EPS) medium, fermentation temperature and fermentation period were optimized. Fermentation of the EPS medium (pH 6·7), containing sucrose at 5% (w/v) and 5% (v/v) inoculum, at 25 ° C resulted in maximum production of HePS (18·38 g l⁻¹) by the isolate in 4 h of fermentation.
Conclusions:  The isolate was found to produce good amount of HePS in just 4 h in a low cost semi-synthetic EPS medium.
Significance and Impact of the Study:  This is the first report on rapid production of HePS by any lactic culture, which can significantly reduce the cost of the EPS.     

Egg hatching inhibition: field evidence for population regulation in a treehole mosquito

ABSTRACT. 1. We investigated the influence of larval density on egg hatching in the treehole mosquito, Aedes triseriatus Say (Diptera: Culicidae) by submerging laboratory-produced egg batches in natural habitats stocked with fixed densities of larvae.
2. We found a significant decrease in hatching rate as larval density increased. The results suggest that inhibited eggs refrain from hatching into environments in which competitive effects of large larvae are potentially severe, and in which cannibalism may occur.
3. The amount of time the egg batches were exposed to experimental conditions (between 2 and 16 days) had no significant influence on hatch rate.     

The assessment of the antibacterial and antifungal activities of aspirin, EDTA and aspirin–EDTA combination and their effectiveness as antibiofilm agents

Aims:  To evaluate the antimicrobial activities of aspirin, EDTA and an aspirin-EDTA (A-EDTA) combination against Pseudomonas aeruginosa , Escherichia coli and Candida albicans in planktonic and biofilm cultures.
Methods and Results:  Minimal inhibitory concentrations (MIC) and minimal biocidal concentrations (MBC) were determined using twofold broth microdilution and viable counting methods, respectively. Aspirin's recorded MIC values ranged from 1·2 to 2·7 mg ml⁻¹. Checkerboard assay demonstrated a synergism in antimicrobial activity upon combination. Aspirin's minimal biofilm eradication concentration values (MBEC) against the established biofilms ranged between 1·35 and 3·83 mg ml⁻¹. A complete eradication of bacterial biofilms was achieved after a 4-h treatment with the A-EDTA combination.
Conclusion:  Both aspirin and EDTA possess broad-spectrum antimicrobial activity for both planktonic and biofilm cultures. Aspirin used at the MBEC for 24 h was successful in eradicating P. aeruginosa , E. coli and C. albicans biofilms established on abiotic surfaces. Moreover, the exposure to the A-EDTA combination (4 h) effected complete bacterial biofilm eradication.
Significance and Impact of the Study:  There is a continuous need for the discovery of new antimicrobial agents. Aspirin and EDTA are 'nonantibiotic drugs', the combination of which can be used successfully to treat and eradicate biofilms established on abiotic surfaces.