Invertebrate biodiversity affects predator fitness and hence potential to control pests in crops

Natural enemies that control pests usually allow farmers to avoid, or reduce, the use of pesticides. However, modern farming practices, that maximize yields, are resulting in loss of biodiversity, particularly prey diversity. Does this matter? Pests continue to thrive, and without alternative prey the predators should, perforce, concentrate their attentions upon the pests.We showed that a diverse diet significantly enhances predator fecundity and survival. Experiments were conducted using common generalist predators found in arable fields in Europe, the carabid beetle Pterostichus melanarius (Coleoptera: Carabidae) and the linyphiid spider Erigone atra (Araneae: Linyphiidae). We tested the hypothesis that mixed species diets were optimal, compared with restricted diets, with respect to parameters such as predator weights, egg weights, numbers of eggs laid, egg development times, egg hatching rates and predator survival. In carabids, an exclusive earthworm diet was as good as mixed diets containing earthworms for egg production and hatching, but less good than such mixed diets for increase in beetle mass and sustained egg laying. For spiders, aphids alone (Sitobion avenae) or with the Collembola Folsomia candida, drastically reduced survival. Aphids plus the Collembola Isotoma anglicana improved survival but only aphids with a mixed Collembola diet maximized numbers of hatching eggs.Predators offered only pests (slugs or aphids) had lowest growth rates and fecundity. We therefore demonstrated that conservation of a diversity of prey species within farmland, allowing predators to exploit a diverse diet, is essential if predators are to continue to thrive in crops and regulate agricultural pests.     

Selectivity underlies the dissociation between seasonal prey availability and prey consumption in a generalist predator

Generalist predators are capable of selective foraging, but are predicted to feed in close proportion to prey availability to maximize energetic intake especially when overall prey availability is low. By extension, they are also expected to feed in a more frequency‐dependent manner during winter compared to the more favourable foraging conditions during spring, summer and fall seasons. For 18 months, we observed the foraging patterns of forest‐dwelling wolf spiders from the genus Schizocosa (Araneae: Lycosidae) using PCR‐based gut‐content analysis and simultaneously monitored the activity densities of two common prey: springtails (Collembola) and flies (Diptera). Rates of prey detection within spider guts relative to rates of prey collected in traps were estimated using Roualdes’ c_st model and compared using various linear contrasts to make inferences pertaining to seasonal prey selectivity. Results indicated spiders foraged selectively over the course of the study, contrary to predictions derived from optimal foraging theory. Even during winter, with overall low prey densities, the relative rates of predation compared to available prey differed significantly over time and by prey group. Moreover, these spiders appeared to diversify their diets; the least abundant prey group was consistently overrepresented in the diet within a given season. We suggest that foraging in generalist predators is not necessarily restricted to frequency dependency during winter. In fact, foraging motives other than energy maximization, such as a more nutrient‐focused strategy, may also be optimal for generalist predators during prey‐scarce winters.     

Invertebrate biodiversity affects predator fitness and hence potential to control pests in crops

Natural enemies that control pests usually allow farmers to avoid, or reduce, the use of pesticides. However, modern farming practices, that maximize yields, are resulting in loss of biodiversity, particularly prey diversity. Does this matter? Pests continue to thrive, and without alternative prey the predators should, perforce, concentrate their attentions upon the pests.We showed that a diverse diet significantly enhances predator fecundity and survival. Experiments were conducted using common generalist predators found in arable fields in Europe, the carabid beetle Pterostichus melanarius (Coleoptera: Carabidae) and the linyphiid spider Erigone atra (Araneae: Linyphiidae). We tested the hypothesis that mixed species diets were optimal, compared with restricted diets, with respect to parameters such as predator weights, egg weights, numbers of eggs laid, egg development times, egg hatching rates and predator survival. In carabids, an exclusive earthworm diet was as good as mixed diets containing earthworms for egg production and hatching, but less good than such mixed diets for increase in beetle mass and sustained egg laying. For spiders, aphids alone (Sitobion avenae) or with the Collembola Folsomia candida, drastically reduced survival. Aphids plus the Collembola Isotoma anglicana improved survival but only aphids with a mixed Collembola diet maximized numbers of hatching eggs.Predators offered only pests (slugs or aphids) had lowest growth rates and fecundity. We therefore demonstrated that conservation of a diversity of prey species within farmland, allowing predators to exploit a diverse diet, is essential if predators are to continue to thrive in crops and regulate agricultural pests.     

Behavioral and ecophysiological responses of a generalist predator to single- and mixed-species diets of different quality

Prey of different quality can be distinguished by their effects on fitness parameters (e.g., survival, growth, development) of the predator. This paper describes a series of experiments with wolf spiders, Schizocosa sp., designed to analyze the behavioral and physiological mechanisms by which these effects are brought about. Schizocosa responded to prey of inferior quality by developing an aversion. The number of prey encounters needed for the aversion to develop depended on the prey type and the quality of alternative prey available. As expected, fewer low-quality prey were accepted if prey of higher quality were available; however, acceptance was also reduced if alternative prey were of even lower quality. An aversion disappeared within a few hours to about 1 day. Consumption by Schizocosa of even small amounts of some “toxic” prey species reduced growth by inhibiting feeding rate and impairing utilization of food derived from prey of higher quality. These results have implications for understanding the interactions of generalist predators in community food webs. Received: 29 July 1998 / Accepted: 1 February 1999     

Impact of intraguild predators on survival of a forest-floor wolf spider

We investigated the impact of variation in densities of a guild of generalist predators on survival of young wolf spiders of the genus Schizocosa. Numbers of other spiders and centipedes were reduced by >80% in fenced 4-m² plots in an experiment that was replicated twice in each of three forest locations. Schizocosa survival during the 1st month was low (<50%) in all three locations, but did not differ between predator-reduction and control plots. By the end of the 1st month, densities of the manipulated predators had converged in control and perturbed treatments, most likely because of reduced per capita mortality from lowered rates of intraguild predation and cannibalism in the experimental treatment. During the 2nd month of the experiment, centipedes and spiders other than Schizocosa again were removed from the experimental plots and, unlike the earlier period, numbers of intraguild predators in the predator-removal treatment remained lower than in control plots. Reducing densities of intraguild predators during the 2nd month improved survival of older juvenile Schizocosa by 75% in two of three locations on the forest floor. In addition to this evidence that intraguild predation can affect older juvenile Schizocosa, survival of Schizocosa during the last half of the experiment was negatively correlated with spatial variation in densities of gnaphosid and ctenid spiders. These two abundant families of cursorial spiders preyed on Schizocosa at a high rate in laboratory trials. Thus, variation in densities of intraguild predators did not influence the youngest Schizocosa, but did influence the survival of older juveniles, most likely due to variations in densities of other cursorial spiders. Received: 21 October 1998 / Accepted: 14 May 1999     

Effects of chronic exposure to a toxic prey in a generalist predator

Abstract.  Generalist predators experience large differences in prey quality, and eating toxic prey may affect their fitness even if the toxic prey constitutes only a small fraction of a mixed diet. Feeding and life history parameters were examined in the wolf spider Pardosa prativaga during continuous exposure to the toxic collembolan Folsomia candida . Spiderlings were divided into a control group fed adult Drosophila melanogaster , a group fed only F. candida , and a mixed diet group fed both types of prey. Folsomia candida reduced survival, developmental stage attained and growth rate of all exposed spiders. Spiders chronically exposed to F . candida increased their consumption of D. melanogaster compared to control spiders. Folsomia candida thus inhibited the utilization of the high-quality food and/or increased the respiration rate. The intake of F . candida remained at a constant low level throughout the experiment, indicating that P. prativaga was unable to develop an absolute aversion against this prey. Half of the control group was also given the mixed diet after a large juvenile instar was attained. These spiders were not affected to the same extent as the newly hatched spiderlings, indicating an ontogenetically increased tolerance. Comparison of laboratory-raised and field-caught spiders confirmed that tolerance to F. candida was size-dependent and thus not an induced response. A differential survival in the mixed diet group, based partly on maternal effects, indicated possible genetic variability in the physiological tolerance to F. candida .     

Influence of prey availability and conspecifics on patch quality for a cannibalistic forager: laboratory experiments with the wolf spider Schizocosa

 Because cannibals are potentially both predator and prey, the presence of conspecifics and alternative prey may act together to influence the rate at which cannibals prey upon each other or emigrate from a habitat patch. Wolf spiders (Lycosidae) are cannibalistic-generalist predators that hunt for prey with a sit-and-wait strategy characterized by changes in foraging site. Little information is available on how both prey abundance and the presence of conspecifics influence patch quality for these cursorial, non-web-building spiders. To address this question, laboratory experiments were conducted with spiderlings and older juveniles of the lycosid genus Schizocosa. The presence of insect prey consistently reduced rates of spider emigration when spiders were housed either alone or in groups. Solitary juvenile Schizocosa that had been recently collected from the field exhibited a median giving-up time (GUT) of 10 h in the absence of prey (Collembola); providing Collembola increased the median GUT to 64 h. For solitary spiders, the absence of prey increased by about fourfold the rate of emigration during the first 24 h. In contrast, for spiders in patches with a high density of conspecifics, the absence of prey increased the 24-h emigration rate by only 1.6-fold. For successful cannibals in the no-prey patches, the presence of conspecifics improved patch quality by providing a source of food. Mortality by cannibalism was affected by both prey availability and openness of the patch to net emigration. In patches with no net emigration, the presence of prey reduced rates of cannibalism from 79% to 57%. Spiders in patches open to emigration but not immigration experienced a rate of cannibalism (16%) that was independent of prey availability. The results of these experiments indicate that for a cannibalistic forager such as the wolf spider Schizocosa, (1) the presence of conspecifics can improve average patch quality when prey are absent, and (2) cannibalism has the potential to be a significant mortality factor under natural field conditions because cannibalism persisted in prey patches that were open to emigration. Received: 12 April 1996 / Accepted: 14 August 1996     

Suitability of woodlice prey for generalist and specialist spider predators: a comparative study

1. Predators select a prey according to its energetic and nutritional composition. Generalist predators avoid, whereas specialists often specialise on well‐defended prey. The aim of this study was to find the suitability of woodlice prey for generalist and specialist predators by comparing their handling efficiency. 2. Laboratory experiments were performed in which specialist and generalist predators were reared on monotypic diets comprising one or other of two woodlice species that differ in their defensive strategies: rollers (Armadillidium) and clingers (Porcellio). A control group was reared on a mixture of arthropods (excluding woodlice). Three spider predators were used that differ in their adaptations to deal with woodlice prey: a woodlice specialist, Dysdera crocata; an oligophagous generalist, Pholcus phalangioides, that also captures woodlice; and a euryphagous generalist, Tegenaria domestica, that does not feed on woodlice. The frequency of capture was recorded and various fitness parameters were measured, namely survival, growth rate, and ontogenetic development. 3. It was found that the specialist, D. crocata, performed best on the Porcellio diet, and similarly well on Armadillidium and mixed diets. The two generalists, P. phalangioides and T. domestica, had poor performance on both woodlice diets but performed well on the mixed diet. 4. The results show that woodlice are unsuitable prey for both oligophagous and euryphagous generalist predators.     

Stable isotope enrichment (δ15N and δ13C) in a generalist predator (Pardosa lugubris, Araneae: Lycosidae): effects of prey quality

Analysis of the natural variations in stable isotope ratios in animal tissue may be a powerful tool to reveal the trophic position and feeding preferences of generalist predators such as lycosid spiders. In the present study, changes in ¹⁵N/¹⁴N and ¹³C/¹²C ratios in the lycosid spider Pardosa lugubris (Araneae: Lycosidae), fed with prey of different quality, were investigated. Experimental food chains included three trophic levels: prey media, prey organism and predator. In order to analyse the time course of stable isotope enrichment, different life stages of P. lugubris feeding on Drosophila melanogaster were studied. The ¹⁵N content of hatchlings of P. lugubris was significantly lower than that of their mothers, indicating the existence of nitrogen pools with different ¹⁵N signatures in female P. lugubris. With duration of feeding and progressing development, age and body weight, the ¹⁵N content in spiderlings increased. Starvation resulted in ¹⁵N and ¹³C enrichment in P. lugubris. Being fed with prey of different quality resulted in varying patterns of stable isotope enrichment. ¹⁵N but not ¹³C content consistently increased when fed on high quality prey (Heteromurus nitidus, D. melanogaster and a mixed diet consisting of H. nitidus and D. melanogaster). In contrast, low quality prey (Rhopalosiphum padi, Folsomia candida and a mixed diet consisting of H. nitidus, D. melanogaster and F. candida) resulted in deviations from postulated patterns of stable isotope enrichment with ¹⁵N content being similar to that of starving spiderlings. When fed on high quality prey, ¹⁵N enrichment of P. lugubris was close to ca. 3‰. It is concluded that the analysis of variations in the natural abundance of stable isotopes is particularly helpful in revealing the trophic structure of terrestrial food webs in which polyphagous feeders predominate, as is the case in litter and soil.     

Capture efficiency and trophic adaptations of a specialist and generalist predator: A comparison

Specialist true predators are expected to exhibit higher capture efficiencies for the capture of larger and dangerous prey than generalist predators due to their possession of specialized morphological and behavioral adaptations. We used an araneophagous spider (Lampona murina) and a generalist spider (Drassodes lapidosus) as phylogenetically related model species and investigated their realized and fundamental trophic niches and their efficacy with respect to prey capture and prey handling. The trophic niche of both species confirmed that Lampona had a narrow trophic niche with a predominance of spider prey (including conspecifics), while the niche of Drassodes was wide, without any preference. DNA analysis of the gut contents of Lampona spiders collected in the field revealed that spiders form a significant part of its natural diet. Lampona captured significantly larger prey than itself and the prey captured by Drassodes. As concerns hunting strategy, Lampona grasped the prey with two pairs of legs possessing scopulae, whereas Drassodes immobilized prey with silk. Lampona possess forelegs equipped with scopulae and a thicker cuticle similar to other nonrelated araneophagous spiders. Lampona fed for a longer time and extracted more nutrients than Drassodes. We show that specialized behavioral and morphological adaptations altogether increase the hunting efficiency of specialists when compared to generalists.     

Food quality of Ephestia eggs,the aphid Rhopalosiphum padi and mixed diet for Orius majusculus

We studied the food quality of the aphid Rhopalosiphum padi to the pirate bug Orius majusculus using Ephestia eggs as high-quality comparison prey. Several performance parameters were tested on individuals that had been reared and maintained on each of the two single-prey diets or on a mixed diet. All fitness parameters were lower in individuals fed aphids only, indicating poor food quality of this prey. Compared with the pure Ephestia egg diet, the mixed diet enhanced teneral mass, while adult survival and female starvation tolerance were negatively affected and all other traits were unaffected. Body protein proportions were constant across diets, whereas lipid proportion was low in the aphid treatment. Preference for aphids was lower following a monotypic aphid diet than when reared on Ephestia eggs or a mixed diet. The results confirm that R. padi is low-quality food for O. majusculus as it is for other generalist predators, even though O. majusculus may contribute significantly to population suppression of the aphid.     

A specialized araneophagic predator's short‐term nutrient utilization depends on the macronutrient content of prey rather than on prey taxonomic affiliation

A specialist predator that has a specialized diet, prey‐specific prey‐capture behaviour and a preference for a particular type of prey may or may not be specialized metabolically. Previous studies have shown that jumping spiders of the genus Portia prey on other spiders using prey‐specific prey‐capture behaviour, prefer spiders as prey to insects and gain long‐term benefits in terms of higher survival and growth rates on spider diets than on insect diets. However, it is unclear whether there are substances uniquely present in spiders on which Portia depends, or, alternatively, spiders and insects all contain more or less the same nutrients but the relative amounts of these substances are such that Portia perform better on a spider diet. These questions are addressed by testing the hypothesis that prey specialization includes metabolic adaptations that allow Portia an enhanced nutrient extraction or nutrient utilization efficiency when feeding on spider prey compared with insect prey. Three groups of Portia quei Zabka are fed either their preferred spider prey or one of two types of flies (Drosophila melanogaster Meigen) that differ in nitrogen and lipid content. Portia quei shows a higher feeding rate of high‐protein flies than of high‐lipid flies and spiders but, after 5 days of feeding, there is no significant difference in growth between treatments, and the diets lead to significant changes in the macronutrient composition of P. quei as a result of variable extraction and utilization of the prey. The short‐term utilization of spider prey is similar to that of high‐lipid flies and both differ in several respects from the utilization of high‐protein flies. Thus, the short‐term nutrient utilization is better explained by prey macronutrient content than by whether the prey is a spider or not. The results suggest that spider prey may have a more optimal macronutrient composition for P. quei and that P. quei does not depend on spider‐specific substances.     

Prey species and prey diet affect growth of invertebrate predators

1. The effects of prey species and leaf age used by prey on performance of two generalist invertebrate predators were studied. The focal plant was Plantago lanceolata , which contains iridoid glycosides.
2. Diet of the herbivorous prey influenced their growth rate.
3. The generalist herbivore ( Vanessa cardui ) and the novel-plant feeder ( Manduca sexta ) contained very low levels of iridoid glycosides in their haemolymph, whereas the specialist ( Junonia coenia ) levels were 50–150-fold higher.
4. Predatory stinkbugs ( Podisus maculiventris ) fed either the novel-plant feeder or the specialist exhibited similar developmental rates. However, stinkbugs ate less of the generalist but grew faster. The growth rate of the stinkbugs was higher when the caterpillar species were raised on the new-leaf powder diet, which contained twice as much protein and iridoid glycosides as the mature-leaf powder diet.
5. Jumping spiders ( Phidippus audax ) ate more mealworms ( Tenebrio molitor ) than specialist J. coenia caterpillars, fed either new- or mature-leaf powder diets, and could not gain weight when fed J. coenia.
6. These results indicate that prey quality was not determined solely by the iridoid glycoside concentration in the diet.     

Maternal Care and Infanticide by Males in Helvibis longicauda (Araneae: Theridiidae)

Maternal care in spiders varies from just the construction of a protective silken structure for the eggs and the selection of a safe site to place them, to a long period of association between the mother and spiderlings. Such extended care may involve the active protection from predators and parasitoids, food regurgitation, the production of trophic eggs and even matriphagy. In this study, we describe extended maternal care in Helvibis longicauda (Theridiidae) and evaluate the effectiveness of maternal protection against predators of eggs and spiderlings. We conducted experiments comparing the frequency of egg sac destruction and mortality of spiderlings in the presence and absence of mothers. We also observed the behaviour of the mother and spiderlings during prey capture events and interactions with possible predators. Helvibis longicauda females guard their egg sacs until the emergence of the young and guard the spiderlings for several instar stages, fighting possible predators, including conspecifics. We found that aggressive behaviour by females increased the survival of both eggs and spiderlings in our experiments. Intruder males were the main source of mortality in the absence of females. The benefits of maternal care for the young also include the acquisition of prey items that are captured, immobilized and pre‐digested by the mother. Effective maternal protection and the extended period of supplying food to juveniles probably contribute to the late dispersal of offspring in H. longicauda.     

Survival strategies of the crab spider Thomisus onustus Walckenaer 1806 (Chelicerata,Arachnida, Thomisidae)

Summary The initial energy supply of emerging spiderlings is relatively meagre, so survival without feeding on insects during a spell of bad weather is limited to a period of a few days or weeks. During our investigations, spiderlings of Thomisus onustus (Arachnida, Thomisidae) were kept on different diets. There was a significant difference in survival rate between spiderlings that were starved or fed on pollen, nectar, or Drosophila. The results showed that pollen and nectar can be a source of energy for spiders for an extensive period. This demonstrates another way in which spiders may survive starvation when insect prey is lacking and thus ensure the survival of a whole population.     

Weak responses to dietary enrichment in a specialized aphid predator

Some ladybeetles are specialist predators of aphids, coccids or other prey, although they often eat a variety of species from their focal prey taxon. In addition, the diet is often supplemented with alternative prey. How larvae of the aphidophagous Coccinella septempunctata L. utilize a non‐aphid alternative prey (fruit‐fly larvae Drosophila melonogaster Meigen) is compared with adequate (i.e. high‐quality) aphid prey provided alone (monotypic diet) or in mixed diets. The alternative prey are presented either nutrient‐enriched (i.e. raised on dog food supplemented medium) or not (raised on pure medium). Ladybird performance (survival, growth and development) is poor on the pure fly larvae diets, and also reduced when given mixed diets compared with the pure aphid diet. Nutrient enrichment of the fly larvae has no positive effects. The physiological background for the differences in food value, as indicated by performance in life‐history parameters, is a strong pre‐ingestive effect (i.e. reduced consumption of fly larvae compared with aphids) and a post‐ingestive effect (i.e. reduced utilization of assimilated larval fly tissue), whereas the assimilation efficiency of the consumed fly larvae is as high as that of aphids. The results show a physiological trade‐off resulting from prey specialization that reduces the possibility of utilizing alternative prey when the availability of aphids is scarce. Connected with this is a high robustness against variation in prey nutrient diversity and composition; the ladybird shows little positive response to dietary mixing (i.e. neither mixing of adequate aphids, nor of aphids and alternative prey) or to nutrient enrichment of prey. This contrasts with the results from generalist predators (spiders), where similar treatments lead to strong effects on life‐history parameters.     

Effects of chlorogenic acid-and tomatine-fed caterpillars on the behavior of an insect predator

If generalist insect predators are a selective force contributing to patterns of feeding specialization by insect herbivores, then predators should be deterred from eating allelochemical-fed prey. The attack and feeding behaviors of naive predators (Podisus maculiventris stinkbugs) reared on control caterpillars (Manduca sexta) fed plain diet were compared to experienced predators reared on caterpillars fed tomato allelochemicals. Tomatine-fed prey were found more quickly by both naive and tomatine-experienced predators, and chlorogenic acid-experienced predators were more stimulated to begin searching for prey. However, experienced predators were less likely to attack both chlorogenic acidfed and tomatine-fed caterpillars than were naive predators. These results indicate that allelochemical-fed prey were easier for predators to locate, but allelochemical-containing prey often deterred predation by experienced predtors.     

Mismatch between dietary requirements for lipid by a predator and availability of lipid in prey

Growth is an important factor in predicting an organism's overall success as an adult. Understanding how abiotic and biotic factors influence body size is key to predicting how environmental changes will impact organisms and predicting optimal behaviors under varying conditions. Food items can vary widely in nutrient content and this variation can affect growth. We tested how the quantity and macronutrient content of live prey affected the growth of juvenile jumping spiders, Phiddipus audax, using 420 spiderlings raised on one of 21 diets. In addition, we measured the nutrient content of prey from the field and compared average prey nutrient content with the diet at which spiders maximized growth. Our results show that the quantity and nutrient content of prey have significant and interacting effects on the growth of an actively hunting predator. In particular, spiders grew larger in body size (tibia/patella length and posterior lateral eye width) and were heavier on diets with higher lipid content. Yet, data on the nutrient content of prey from the field shows that most potential prey have high protein and low lipid content in their bodies. This apparent mismatch between spider nutritional requirements and prey nutrient availability likely presents a challenge to spiders seeking to maximize growth in the field. Several hypotheses for how these predators overcome this challenge to reach the large body sizes observed in the field are discussed.     

Spider predation on a mirid pest in Japanese rice fields

Spiders are common generalist predators, and understanding their potential in biological control is important for the development of integrated pest management programs. In this study, predation by three groups of spiders on the mirid bug Stenotus rubrovittatus (Hemiptera: Miridae) in rice paddies was investigated using DNA-based gut-content analysis. A laboratory feeding study revealed that the detection half-lives of bug DNA in the spider gut at 25 °C was 3.4 days for Lycosidae and 1.5 days for Tetragnathidae. Individual spider predation on the mirid bug was investigated by detecting DNA of prey in field-collected spiders. In total, 1199 spiders were assayed from three spider groups: Pirata subpiraticus (Lycosidae), Tetragnatha spp. (Tetra-gnathidae), and Pachygnatha clercki (Tetra-gnathidae), which each differ in their preferred microhabitat as well as their predatory habits. Detection rates of prey DNA in spiders increased significantly with the density of prey across all spider groups. P. subpiraticus and Tetragnatha spp. predation showed a better fit to a saturated response curve to increasing prey density, while P. clercki showed a simple linear relationship with prey density. Densities of alternative prey species did not affect the detection rates of mirids. These results suggest that predation on pests by generalist predators in an agroecosystem is affected not only by prey abundance but also by predator preference for specific prey. Predator preference is therefore an important factor to consider when estimating the role of natural enemies as biological control agents.     

An order-specific monoclonal antibody to Diptera reveals the impact of alternative prey on spider feeding behavior in a complex food web

Generalist predators have the capacity to restrict pest population growth, especially early in the season before densities increase. However, their polyphagous feeding habits sometimes translate into reduced pest consumption when they target alternative prey. An order-specific monoclonal antibody was developed to examine the strength of trophic connections between Diptera, a major category of non-pest prey, and linyphiid spiders in alfalfa. We report the development and characterization of a monoclonal antibody with order-level specificity to Diptera. This antibody elicited strong absorbance to 22 Diptera from 13 families, no false-positive reactivity to non-dipteran invertebrates, and antigen detection periods following prey consumption that were comparable between spiders. Over 900 field-collected females of the linyphiid spiders Erigone autumnalis and Bathyphantes pallidus were screened for Diptera antigen. Significantly more B. pallidus screened positive for Diptera (40%) compared to E. autumnalis (16%), indicating differential reliance on these prey. In parallel with the collection of spiders for gut-content analysis, prey availability was estimated at web sites. The two spiders exhibited different feeding responses to prey availability. Consumption of Diptera by B. pallidus was strongly correlated with Diptera abundance whilst the availability of other potential prey did not influence predation rates. Conversely, E. autumnalis did not prey upon Diptera in proportion to availability, but increased Collembola activity-density reduced dipteran consumption. Integration of molecular gut-content analysis with precise sampling of prey demonstrated how two closely related linyphiid spiders exhibit different feeding responses to the availability of prey under natural field conditions. Elucidating the feeding preferences of natural enemies is critical to effective incorporation of biological control by generalist predators in the management of agricultural pests.