Spatiotemporal Analysis of Predation by Carabid Beetles (Carabidae) on Nematode Infected and Uninfected Slugs in the Field

The dynamics of predation on parasites within prey has received relatively little attention despite the profound effects this is likely to have on both prey and parasite numbers and hence on biological control programmes where parasites are employed. The nematode Phasmarhabditis hermaphrodita is a commercially available biological agent against slugs. Predation on these slugs may, at the same time, result in intraguild predation on slug-parasitic nematodes. This study describes, for the first time, predation by carabid beetles on slugs and their nematode parasites on both spatial and temporal scales, using PCR-based methods. The highest nematode infection levels were found in the slugs Deroceras reticulatum and Arion silvaticus. Numbers of infected slugs decreased over time and no infected slugs were found four months after nematode application. The density of the most abundant slug, the invasive Arion vulgaris, was positively related to the activity-density of the carabid beetle, Carabus nemoralis. Predation on slugs was density and size related, with highest predation levels also on A. vulgaris. Predation on A. vulgaris decreased significantly in summer when these slugs were larger than one gram. Predation by C. nemoralis on slugs was opportunistic, without any preferences for specific species. Intraguild predation on the nematodes was low, suggesting that carabid beetles such as C. nemoralis probably do not have a significant impact on the success of biological control using P. hermaphrodita.     

The rhabditid nematode Phasmarhabditis hermaphrodita as a potential biological control agent for slugs

A nematode, Phasmarhabditis hermaphrodita, known to be associated with slugs but not previously thought to be parasitic, was shown to be a parasite capable of killing the pest slug Deroceras reticulatum. The parasite infects slugs in the area beneath the mantle surrounding the shell, causing a disease with characteristic symptoms, particularly swelling of the mantle. Infection leads to death of the slug, usually between seven and 21 days afterwards. The nematode then spreads and multiplies in the cadaver. In an experiment where individual D. reticulatum were exposed to different numbers of P. hermaphrodita, a significant positive relationship was found between nematode dose and slug mortality. In two experiments on host range, the nematode was found to infect and kill all pest slug species tested: Deroceras caruanae, Arion distinctus, Arion silvaticus, Arion intermedius, Arion ater, Tandonia sowerbyi and T. budapestensis, in addition to D. reticulatum.     

Effect of predator size and temperature on the predation of Deroceras reticulatum (Muller) (Mollusca) by carabid beetles

Twenty-one carabid species were used in a laboratory study to determine the effect of beetle size and temperature on the predation of one day old Deroceras reticulatum (Muller) slugs. The beetles were caught in arable land at the study site. None of the five small-sized beetle species killed slugs, whereas five of the seven medium-sized beetle species and eight of the nine large-sized beetle species killed slugs. Four of the most abundant beetle species at the study site were used to assess the effect of temperature on the predation of 1-day-old slugs. There were significant differences in the proportion of beetles predating slugs at the different temperatures. Regression analysis showed that there was a significant increase in the proportion of Pterostichus madidus (Fabricius) and Harpalus rufipes (Degeer) beetles predating slugs as temperature increased. Nebria brevicollis (Fabricius) inflicted its highest level of slug mortality at 8°C and its larvae were the only predators to significantly increase slug mortality at 4°C, the lowest experimental temperature. Harpalus aeneus (Fabricius) inflicted significant levels of slug mortality at 20°C, the highest experimental temperature. It appeared that the predatory activity of these beetle species at the five experimental temperatures reflected their annual activity periods in the field.     

Molluscicidal and slug-repellent properties of anaerobically digested organic matter

Slug problems in arable crops and vegetables have increased drastically during the past few decades. Observations on slug damage to oilseed rape suggested that fresh, anaerobically digested organic material from a biogas production plant is molluscicidal. To find out whether digested matter can be used for the control of agricultural pest slugs, a series of exeriments were carried out. The laboratory experiments demonstrated strong mollusc repellent and molluscicidal effects of digested organic matter against the three most important pest slugs of Switzerland, Arion lusitanicus, A. distinctus and Deroceras reticulatum. The effects were restricted to fresh digested matter and were rapidly lost when the material was stored, and also after application in the field. In the field experiment, fresh digested matter greatly reduced slug damage to lettuce in comparison with the untreated plots. At present, the chemical nature of the molluscicidal compound(s) in digested matter is unknown, but environmental pollutants such as heavy metals can be ruled out. Current research aims at a new formulation which is easier to apply and has a longer-lasting molluscicidal or slug-repellent effect, and at optimising the dosage and number of applications.     

Predation and prey size choice by the carabid beetle Pterostichus melanarius (Coleoptera: Carabidae): the dangers of extrapolating from laboratory to field

The impact of predation by the generalist carabid beetle Pterostichus melanarius (Illiger) on populations of the field slug Deroceras reticulatum (Müller), and the effects of prey size on the predator-prey interaction, were measured under semi-field conditions. It was hypothesized that environmental heterogeneity would lead to very different patterns of comparative mortality than might be deduced from size choice experiments conducted in the laboratory. Results from outdoor mini-plots, emulating conditions in a field of wheat, demonstrated that P. melanarius significantly reduced numbers of slugs from all size classes, with no apparent preferences. This was in marked contrast to results from earlier laboratory studies, where this beetle fed preferentially on the smallest slugs. The slugs in the mini-plots ranged in size from 2-100 mg and the numbers in the mini-plot reflected the size frequency distribution in the field. Beetles in mini-plots containing high densities of slugs increased significantly in weight, in contrast to beetles in mini-plots with low slug density or no added slugs, which did not. Enzyme-linked immunosorbent assays (ELISA), using anti-slug monoclonal antibodies, showed that where there was a higher density of slugs there was more slug protein in the guts of the beetles. It was concluded that environmental heterogeneity probably provided a greater number and diversity of refugia for smaller than for larger slugs, counteracting laboratory-measured size preferences measured in arenas without refugia. These results have implications for a range of ecological studies involving inter- and intra-specific prey size choice, and emphasize the dangers of extrapolating from the laboratory to the field.     

Control of slugs with contact-action molluscicides

A number of iron and aluminium compounds were compared as contact-acting poisons against the slug Deroceras reticulatum, by confining slugs on treated surfaces of glass and wet soil in laboratory tests. Simple metal salts were effective poisons when applied to a glass surface, but were rapidly inactivated when applied to wet soil. Chelation of both metals with organic ligands retarded the rate of attenuation on wet soil. In a field trial D. reticulatum and Arion spp. slugs were killed by a broadcast application of the chelate iron (III) 2,4-pentanedionate applied at 40 kg a.i. ha^-1.     

THE IMPORTANCE OF MUCUS AS A DEFENCE AGAINST CARABID BEETLES BY THE SLUGS ARION FASCIATUS AND DEROCERAS RETICULATUM

The effect of mucus exudation on the survival of Arion fasciatusand Deroceras reticulatum was studied by exposing mechanicallystimulated and control slugs to Carabidae beetles for 24 hours.A light stimulation, lasting three minutes, exhausted the copiousflow of mucus for one day. A generalist, Pterostichus niger,significantly preferred stressed D. reticulatum over controlones. Similarly, P. niger exclusively ate stressed individualsof A. fasciatus. Two beetles which specialize in gastropods,Cychrus caraboides and Carabus violaceus, consumed an equalnumber of stressed and control D. reticulatum and A. fasciatus.The susceptibility of the slug species to predation was different:for each beetle species, the proportion of available D. reticulatumpreyed upon was significantly higher than that of A. fasciatus.The differences in the behaviour of A. fasciatus and D. reticulatumin defending themselves against attacks is described. The mainreason why specialist beetles are able to hunt slugs successfullyis their ability to prevent the slugs from exuding large amountsof mucus. This may succeed by different means: C. violaceustargeted their killing strokes against the posterior part ofthe mantle while C. caraboides hit the head of the slug. Inboth case the strokes seemed to paralyze the slugs. (Received 31 March 1993; accepted 1 October 1993)     

Cold tolerance and freeze-induced glucose accumulation in three terrestrial slugs

Cold tolerance and metabolic responses to freezing of three slug species common in Scandinavia (Arion ater, Arion rufus and Arion lusitanicus) are reported. Autumn collected slugs were cold acclimated in the laboratory and subjected to freezing conditions simulating likely winter temperatures in their habitat. Slugs spontaneously froze at about -4 °C when cooled under dry conditions, but freezing of body fluids was readily induced at -1 °C when in contact with external ice crystals. All three species survived freezing for 2 days at -1 °C, and some A. rufus and A. lusitanicus also survived freezing at -2 °C. (1)H NMR spectroscopy revealed that freezing of body fluids resulted in accumulation of lactate, succinate and glucose. Accumulation of lactate and succinate indicates that ATP production occurred via fermentative pathways, which is likely a result of oxygen depletion in frozen tissues. Glucose increased from about 6 to 22 μg/mg dry tissue upon freezing in A. rufus, but less so in A. ater and A. lusitanicus. Glucose may thus act as a cryoprotectant in these slugs, although the concentrations are not as high as reported for other freeze tolerant invertebrates.     

Effects of seed-bed conditions on slug numbers and damage to winter wheat in a clay soil

In a field experiment three levels of consolidation were combined with three seed-bed tilths, to produce nine different types of seed-bed. The soil properties of each combination of tilth and consolidation were measured and effects on the numbers and biomass of slugs in the top 10 cm of soil and on damage to wheat seeds and seedlings were assessed. Eight species of slug were present, with three species predominating (Deroceras reticulatum, Arion distinctus and Arion subfuscus). Between 3% and 33% of seeds and seedlings were killed by slugs (recorded at Growth Stage 12). Contrary to expectation, most damage occurred on consolidated seed-beds of fine or medium tilth, least on loose seed-beds of the same texture; intermediate damage occurred on cloddy seed-beds, where consolidation had little effect. The level of damage occurring on different types of seed-bed was directly related to the biomass of slugs in the top 10 crn of soil, and was inversely related to depth of sowing and the percentage of fine soil in the seed-bed. These three factors together accounted for 94–97% of the variance in slug damage. Consolidation was associated with increased slug damage probably because of its effects on these three factors: slug biomass was greater and seed was at shallower depth in consolidated than loose seed-beds and consolidation, whether before or after drilling, failed to break down clods of soil into finer particles. This is the first experimental evidence in the field of the effects of seedbed conditions on slug numbers and damage to winter wheat and represents a significant step towards forecasting and avoiding slug damage in this crop.     

The effects of crop development upon slug distribution and control by Abax parallelepipedus (Coleoptera: Carabidae)

The polyphagous carabid beetle Abax parallelepipedus has been shown to be capable of controlling slugs within a lettuce crop in previous studies. This report describes experiments undertaken in plots within a polythene tunnel. Field slugs, Deroceras reticulatum, were introduced to plots containing either large or small lettuce plants, at 30 slugs per plot. The plots were further subdivided into those with or without six beetle predators. Slug numbers were assessed after two weeks, and were found to be affected by plant size, both with and without the presence of predators. These two factors had an additive effect, with both small plant size and the presence of beetles causing significant reductions in slug numbers. Plots with beetles contained either males or females, and females were shown to be significantly better at reducing slug numbers than males, particularly within plots of small plants. The four principal treatments (combinations of two plant sizes and the presence/absence of beetles) generated four distinct patterns of slug distribution within the plots. The potential of the residual slug populations to inflict further damage was measured in a subsequent crop of seedling lettuces. It appeared that the beetles were generally incapable of capturing slugs within large lettuce plants, but were effective predators at soil level. Possible reasons for the survival of fewer slugs in plots with small plants, both with and without the presence of predators, are discussed.     

Spatial sampling to detect slug abundance in an arable field

Overuse of molluscicides by farmers in arable systems can lead to environmental and product contamination. Here we assess a simple and inexpensive surface trapping method for monitoring populations of slugs (Deroceras reticulatum and Arion intermedius). This method was biased against small slugs, and against A. intermedius, when compared to direct soil sampling. Regression was used to model the relationship between the results of surface trapping and soil sampling methods. Spatial Analysis by Distance IndicEs (SADIE) algorithms were used to describe the spatial relationships between the two sets of samples. Using both traditional statistical methods and spatial statistics, the spatial information collected from surface traps was sufficient to identify patches and gaps in slug numbers and possibly to allow the spot application of slug control, and thus provide land managers who experience slug damage with a way of reducing molluscicides use, whilst maintaining slug control. Further improvements and applications of the model are discussed.     

Daily activity rhythm and use of shelter in the slugs Deroceras reticulation and Arion distinctus under laboratory conditions

Daily activity of slugs, Deroceras reticulatum and Arion distinctus, was studied in the laboratory in relation to the presence of shelters so as to define optimal conditions for using traps to forecast crop damage. Under constant temperature and humidity, activity of slugs began with the lights off. Its arrest occurred after a period of activity with the lights on. Slugs rested most frequently under a shelter on bare earth and rarely used the same shelter for more than two consecutive days. When food was placed under the shelter, the number of slugs found under the shelter was more constant throughout the 24 h period, but lower during diurnal rest. The presence of 4% methiocarb pellets under the shelter led to an increased occupation by slugs, due to the poisoning of some of them. Acts of aggression were more numerous in A. distinctus than in D. reticulatum. In D. reticulatum the smallest slug was the least aggressive and suffered more attacks from the other slugs. In A. distinctus the numbers of attacks suffered or provoked varied little with the size of the slug, but the smallest slug showed less interactions with other slugs.     

A Species-specific Monoclonal Antibody System for Detecting the Remains of Field Slugs, Deroceras reticulatum (Muller) (Mollusca: Pulmonata), in Carabid Beetles (Coleoptera: Carabidae)

The field slug, Deroceras reticulatum (Muller), is probably the most damaging and widespread species of slug, causing severe economic losses in a broad range of crops in temperate zones throughout the world. Investigations into the role of predation in the population dynamics of this species required a biochemical system which was capable of identifying the remains of this slug in the crop contents of predators, and distinguishing them from those of other molluscs. A monoclonal antibody was developed (IgM isotype). This was capable of separating, in an enzyme-linked immunosorbent assay, D. reticulatum from the related D. caruanae (Pollonera) and all molluscs and other invertebrates tested, with the unexpected exceptions of New Zealand flatworms, Artioposthia triangulata (Deny) and the millipede Polymicrodon polydesmoides (Leach). Characterization of the antibody and assay demonstrated that the system could clearly detect as little as 11.6 ng of D. reticulatum protein in 200 mu l of buffer. Slug remains could be identified as such in the crops of the carabid predator Pterostichus melanarius Illiger for 38.1 h, while the antibody-antigen reaction declined to half of that measured immediately following consumption, after 12.9 h. A practical and highly sensitive system was therefore developed, using the first species-specific monoclonal antibody available for the investigation of predation on slugs.     

The significance of facultative scavenging in generalist predator nutrition: detecting decayed prey in the guts of predators using PCR

Gut-content analyses using molecular techniques are an effective approach to quantifying predator-prey interactions. Predation is often assumed but scavenging is an equally likely route by which animal DNA enters the gut of a predator/scavenger. We used PCR (polymerase chain reaction) to detect scavenged material in predator gut homogenates. The rates at which DNA in decaying slugs (Mollusca: Pulmonata) and aphids (Homoptera: Aphididae) became undetectable were estimated. The detectability of DNA from both carrion types in the guts of the generalist predator Pterostichus melanarius (Coleoptera: Carabidae) was then determined. The effects of carrion age and weight, as well as beetle sex, on detection periods, were quantified. Laboratory trials measured prey preference of beetles between live and decaying prey. Further experiments measured, for the first time, feeding by P. melanarius on dead slugs and aphids directly in the field. In both field and laboratory, P. melanarius preferentially fed on dead prey if available, but preference changed as the prey became increasingly decayed. Disappearance rates for slug carrion in wheat fields and grasslands were estimated and P. melanarius was identified as the main scavenger. Comparison of the retention time for dead slugs in the field, with the detection period for decaying slug material in the guts of the predators, showed that PCR-based techniques are not able to distinguish between predated and scavenged food items. This could potentially lead to overestimation of the impact of predation on slugs (and other prey) by carabids. Possible implications of facultative scavenging by invertebrate predators for biocontrol and food-web research are discussed.     

Effects of metaldehyde and methiocarb on feeding preferences and survival of a slug predator (Pterostichus melanarius (F.): Carabidae,Pterostichini)

The potential for reducing slug populations in crops through predation by generalist carabid beetles is well documented. However, few studies have considered interactions between biological and chemical control agents of slugs. Laboratory assays supported previous findings that the consumption of metaldehyde by slugs (Deroceras reticulatum) leads to increased duration of feeding bouts by carabid beetles (such as Pterostichini) on sub‐lethally affected individuals. However, a similar effect was not found for Pterostichus melanarius exposed to slugs fed on the other widely applied pelleted molluscicide formulation (methiocarb). Examination of beetle survival after consumption of slugs containing molluscicides demonstrated the strong biocidal properties of methiocarb, whereas metaldehyde consumption (ingested through slug predation) did not differ from control slugs killed by freezing. Beetle avoidance of slugs containing a more toxic molluscicide compound and the interaction between slug mucus production and beetle attack rates are discussed.     

SLUG DAMAGE AND NUMBER OF SLUGS (GASTROPODA: PULMONATA) IN WINTER WHEAT IN FIELDS WITH SOWN WILDFLOWER STRIPS

Previous observations have shown that slugs found favourableconditions for reproduction in wildflower strips and that theycaused high damage to oilseed rape adjacent to the strips. Inthe current study slug numbers and damage were estimated inwildflower strips and at different distances from the stripsinto winter wheat crops, from 1994 to 1996. Slug damage wasestimated using an index of defoliation and slug numbers andactivity were measured using wheat bran traps. Investigationsbegan when winter wheat seedlings emerged and lasted for fiveweeks. Slug damage was never severe in any of the fields studied.In most fields, slug damage was higher close to the wildflowerstrips than at greater distances from the strips. The decliningslug damage with increasing distances from the wildflowers stripswas related to the distribution pattern of juvenile Arion lusitanicus,suggesting that this species was responsible for the higherdamage near the strips. Other slug species (Deroceras reticulatum,Arion fasciatus, Deroceras laeve) were more or less evenly distributedover the field or were almost entirely confined to the wildflowerstrips. Results showed that winter wheat was not at risk fromslug feeding in spite of the establishment of wildflower stripsand that therefore the application of molluscicide along thestrips is unnecessary. (Received 23 June 1997; accepted 17 October 1997)     

Biocontrol of Slugs in Protected Lettuce Using the Rhabditid Nematode Phasmarhabditis hermaphrodita

In two experiments, the rhabditid nematode Phasmarhabditis hermaphrodita, a parasite of slugs, was cultured in vitro and applied as a drench to soil at four dose rates (3 108, 1 109, 3 109 and 1 1010 ha-1) 1 or 4 days before planting lettuce seedlings in a polythene tunnel. The effects of the four nematode doses on slug damage during the first 3 weeks after planting and on the numbers of slugs found within and below lettuce plants at harvest were measured. Results were then compared with untreated plots and with plots where methiocarb pellets were applied at the recommended field rate. In the first experiment, methiocarb pellets significantly reduced the percentage of plants damaged by slugs, but the nematode did not. In the second experiment, methiocarb pellets and the second highest dose of nematodes significantly reduced the percentage of plants damaged by slugs. The different effect of the nematode in the two experiments may have resulted from differences in the timing of nematode application and/or differences in the pattern of slug damage between experiments. At the end of the first experiment, the highest two doses of nematodes and methiocarb pellets had significantly reduced the number of slugs found within lettuce plants at harvest and on the soil surface below the plants. At the end of the second experiment, analysis of variance showed no significant effects of any treatment on slug numbers or biomass, but regression analysis showed significant negative relationships between nematode dose and total slug numbers, numbers of Arion ater agg. and biomass of Deroceras reticulatum. In both experiments, increasing nematode dose significantly reduced the numbers of slugs found contaminating the harvested lettuce. At the end of the second experiment, the mean weight of individuals of A. ater agg. increased with rising nematode dose.     

Tracking predator density dependence and subterranean predation by carabid larvae on slugs using monoclonal antibodies

1. Subterranean carabid larvae are more numerous than surface‐active adults, yet very little is known about their ecological significance, dietary preferences or ability to regulate populations of prey species, particularly pests. Part of the reason for this is that predator–prey interactions beneath the soil are almost impossible to observe. 2. Extensive field studies have shown that adult Pterostichus melanarius (Illiger) can affect the temporal and spatial dynamics of their slug prey. However, if larvae too are feeding on slugs, this could radically affect overall predator–prey dynamics. 3. We tested the hypotheses that P. melanarius larvae would kill and consume two slug species, Deroceras reticulatum Müller and Arion intermedius Normand, under laboratory and semi‐field conditions, and that there would be no significant difference in rates of predation on these slug species. 4. A new monoclonal antibody was developed that was capable of detecting the presence of slug proteins in the guts of P. melanarius larvae. 5. Pterostichus melanarius larvae killed both A. intermedius and D. reticulatum in the laboratory, feeding to a greater extent, and growing more rapidly, on the latter. The larvae were equally effective at reducing numbers of both slug species in a crop of wheat grown in semi‐field mini plots, but predation was affected by density‐dependent intra‐specific competition amongst the beetle larvae. 6. Future modelling of the dynamic interactions between carabids and slugs will need to take into account predation by larvae.     

The role of invertebrates in seedling establishment in a heterocarpic plant,Atriplex sagittata

Seed heteromorphism is a marked character of many Chenopodioideae (Amaranthaceae). Seed morphs differ in dormancy, germination and seedling biology, but differences in their predation have not yet been studied. Atriplex sagittata produces small black dormant and large brown non‐dormant seeds. In this study, the timing of seed release and seedling establishment were ascertained, and their consumption by invertebrates (carabids, isopods and slugs) was studied. Seeds dispersed in the autumn passed the winter on the ground surface, protected from invertebrate predation by low temperatures. In the following vegetative season, ungerminated black seeds exposed to predation on ground surface were preferred by a large carabid species, Pseudoophonus rufipes. Some black and all brown seeds escaped predation by germinating in early spring. The seedlings were little endangered by carabids and isopods but were preferred by an invasive slug, Arion vulgaris, the feeding of which can exterminate seedlings at places in which slugs are abundant. Invertebrate predation is important factor of seed and seedling mortality of A. sagittata and seed heteromorphism modifies its intensity and timing.     

DOES TANDONIA BUDAPESTENIS (MOLLUSCA: PULMONATA) CONTAIN TOXINS? EVIDENCE FROM FEEDING TRIALS WITH THE SLUG PREDATOR PTEROSTICHUS MELANARIUS (COLEOPTERA: CARABIDAE)

Slugs are known to be killed and consumed by a range of invertebrateand vertebrate predators in the field. Carabid beetles (Coleoptera:Carabidae) in particular are major natural enemies of slugs,and have been shown to be capable of controlling certain speciesin a crop environment. This paper reports experiments to investigatethe effects, on the carabid Pterostichus melanarius, of feedingon the milacid slug Tandonia budapestensis. The slugs provedto be toxic, 50% of the beetles dying within two days. Mortalityof beetles fed on two other species of slug, Deroceras reticulatumand Arion distinctus, was not significantly different from thatfor beetles fed upon a control prey (Calliphora vomitoria).These results represent the only known case of a European slugproving to be toxic to potential predators, and is one of avery small number of reported instances of possible toxicityamongst terrestrial gastropods. The possibility that the orangeline down the dorsal keel of T. budapestensis may act as warningcoloration to birds is discussed. (Received 4 December 1996; accepted 5 March 1997)