Prey DNA detection success following digestion by intraguild predators: influence of prey and predator species

Intraguild predation (IGP) has been increasingly recognized as an important interaction in ecological systems over the past two decades, and remarkable insights have been gained into its nature and prevalence. We have developed a technique using molecular gut-content analysis to compare the rate of IGP between closely related species of coccinellid beetles (lady beetles or ladybirds), which had been previously known to prey upon one another. We first developed PCR primers for each of four lady beetle species: Harmonia axyridis, Coccinella septempunctata, Coleomegilla maculata and Propylea quatuordecimpunctata. We next determined the prey DNA detection success over time (DS(50) ) for each combination of interacting species following a meal. We found that DS(50) values varied greatly between predator-prey combinations, ranging from 5.2 to 19.3 h. As a result, general patterns of detection times based upon predator or prey species alone are not discernable. We used the DS(50) values to correct field data to demonstrate the importance of compensation for detection times that are specific to particular predator-prey combinations.     

Detecting frogs as prey in the diets of introduced mammals: a comparison between morphological and DNA‐based diet analyses

Amphibians are currently the most threatened group of vertebrates worldwide, and introduced fauna play a major role in their decline. The control of introduced predators to protect endangered species is often based on predation rates derived from diet studies of predators, but prey detection probabilities using different techniques are variable. We measured the detectability of frogs as prey, using morphological and DNA‐based diet analyses, in the stomachs and faeces of four mammal species that have been introduced to many areas of the world. Frogs (Litoria raniformis) were fed to rats (Rattus norvegicus and R. rattus), mice (Mus musculus) and hedgehogs (Erinaceus europaeus). DNA‐based analysis outperformed morphological analysis, increasing the prey detection rate from 2% to 70% in stomachs and from 0% to 53% in faeces. In most cases, utilizing either stomachs or faeces did not affect the success of prey DNA detection; however, using faeces extended the detectability half‐life from 7 to 21 h. This study is the first to measure prey DNA detection periods in mammalian stomachs, and the first to compare prey DNA detection periods in the stomachs and faeces of vertebrates. The results indicate that DNA‐based diet analysis provides a more reliable approach for detecting amphibians as prey and has the potential to be used to estimate the rate of predation by introduced mammals on endangered amphibians.     

Specific detection of the floodwater mosquitoes Aedes sticticus and Aedes vexans DNA in predatory diving beetles

Floodwater mosquitoes (Diptera: Culicidae) are associated with periodically flooded wet meadows, marshes, and swamps in floodplains of major rivers worldwide, and their larvae are abundant in the shallow parts of flooded areas. The nuisance caused by the blood‐seeking adult female mosquitoes motivates mosquito control. Larviciding with Bacillus thuringiensis israelensis is considered the most environmentally safe method. However, some concern has been raised whether aquatic predatory insects could be indirectly affected by this reduction in a potential vital prey. Top predators in the temporary wetlands in the River Dalälven floodplains are diving beetles (Coleoptera: Dytiscidae), and Aedes sticticus and Ae. vexans are the target species for mosquito control. For detailed studies on this aquatic predator–prey system, we developed a polymerase chain reaction (PCR) assay for detection of mosquito DNA in the guts of medium‐sized diving beetles. Primers were designed for amplifying short mitochondrial DNA fragments of the cytochrome C oxidase subunit I (COI) gene in Ae. sticticus and Ae. vexans, respectively. Primer specificity was confirmed and half‐life detectability of Ae. sticticus DNA in diving beetle guts was derived from a feeding and digestion experiment. The Ae. sticticus DNA within diving beetle guts was detected up to 12 h postfeeding, and half‐life detectability was estimated to 5.6 h. In addition, field caught diving beetles were screened for Ae. sticticus and Ae. vexans DNA and in 14% of the diving beetles one or both mosquito species were detected, showing that these mosquito species are utilized as food by the diving beetles.     

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.     

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.     

Intranuclear verrucomicrobial symbionts and evidence of lateral gene transfer to the host protist in the termite gut

In 1944, Harold Kirby described microorganisms living within nuclei of the protists Trichonympha in guts of termites; however, their taxonomic assignment remains to be accomplished. Here, we identified intranuclear symbionts of Trichonympha agilis in the gut of the termite Reticulitermes speratus. We isolated single nuclei of T. agilis, performed whole-genome amplification, and obtained bacterial 16S rRNA genes by PCR. Unexpectedly, however, all of the analyzed clones were from pseudogenes of 16S rRNA with large deletions and numerous sequence variations even within a single-nucleus sample. Authentic 16S rRNA gene sequences were finally recovered by digesting the nuclear DNA; these pseudogenes were present on the host Trichonympha genome. The authentic sequences represented two distinct bacterial species belonging to the phylum Verrucomicrobia, and the pseudogenes have originated from each of the two species. Fluorescence in situ hybridization confirmed that both species are specifically localized, and occasionally co-localized, within nuclei of T. agilis. Transmission electron microscopy revealed that they are distorted cocci with characteristic electron-dense and lucent regions, which resemble the intranuclear symbionts illustrated by Kirby. For these symbionts, we propose a novel genus and species, ‘Candidatus Nucleococcus trichonymphae'' and ‘Candidatus Nucleococcus kirbyi''. These formed a termite-specific cluster with database sequences, other members of which were also detected within nuclei of various gut protists, including both parabasalids and oxymonads. We suggest that this group is widely distributed as intranuclear symbionts of diverse protists in termite guts and that they might have affected the evolution of the host genome through lateral gene transfer.     

Resolving the predator first paradox: Arthropod predator food webs in pioneer sites of glacier forelands

Primary succession on bare ground surrounded by intact ecosystems is, during its first stages, characterized by predator‐dominated arthropod communities. However, little is known on what prey sustains these predators at the start of succession and which factors drive the structure of these food webs. As prey availability can be extremely patchy and episodic in pioneer stages, trophic networks might be highly variable. Moreover, the importance of allochthonous versus autochthonous food sources for these pioneer predators is mostly unknown. To answer these questions, the gut content of 1,832 arthropod predators, including four species of carabid beetles, two lycosid and several linyphiid spider species caught in early and late pioneer stages of three glacier forelands, was screened molecularly to track intraguild and extraguild trophic interactions among all major prey groups occurring in these systems. Two‐thirds of the 2,310 identified food detections were collembolans and intraguild prey, while one‐third were allochthonous flying insects. Predator identity and not successional stage or valley had by far the strongest impact on the trophic interaction patterns. Still, the variability of prey spectra increased significantly from early to late pioneer stage, as did the niche width of the predators. As such the structure of pioneer arthropod food webs in recently deglaciated Alpine habitats seems to be driven foremost by predator identity while site and early successional effects contribute to a lesser extent to food web variability. Our findings also suggest that in these pioneer sites, predatory arthropods depend less on allochthonous aeolian prey but are mainly sustained by prey of local production.     

Detection of Bemisia tabaci remains in predator guts using a sequence-characterized amplified region marker

Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae) B biotype is an invasive species (biotype) in China. In order to understand the role that native natural enemies might play in its control, techniques were developed for detecting B. tabaci DNA within the gut of predators. A species-specific DNA fragment, ca. 350 bp, was identified by random amplified polymorphic DNA analysis. This fragment was absent in other closely related or co-occurring prey species, cotton, and other select predator species. After cloning and sequencing the fragment, one pair of sequence-characterized amplified region (SCAR) primers was developed, which amplified a single band of 240 bp. Specificity tests performed with the primers showed the presence of the 240-bp band for B. tabaci in all developmental stages and both sexes, in adult Propylaea japonica (Thunberg) (Coleoptera: Coccinellidae) fed on B. tabaci nymphs in the laboratory, and in predators collected in cotton fields. Following consumption of a single red-eyed B. tabaci nymph, prey DNA was detectable in 100% of P. japonica at t = 0, decreasing to 20% after 12 h of digestion, and no B. tabaci DNA detected at t = 24 h. In total, we analyzed the gut contents of 185 field-collected predators, representing four different orders. All nine field-collected predator species (namely, P. japonica, Harmonia axyridis, Scymnus hoffmanni, Coccinella septempunctata, Orius sauteri, Chrysopa pallens, Chrysopa formosa, Erigonnidium graminicolum, and Neoscona doenitzi) contained DNA from B. tabaci and are assumed predators of this pest insect. Overall, the B. tabaci was eaten by more than 50% of field-collected predator individuals, including larvae of the coccinellids (P. japonica and H. axyridis) and lacewings (C. pallens and C. formosa) and adults of O. sauteri and the spiders (E. graminicolum and N. doenitzi). There was a trend of a higher percentage of larval than adult coccinellids and lacewings that preyed on B. tabaci in the field. This study provides a framework for the future use of molecular gut content analysis in arthropod conservation ecology and food web research, with considerable potential for quantifying threats to invasive or endemic pest species in China and elsewhere.     

Prey selection by linyphiid spiders: molecular tracking of the effects of alternative prey on rates of aphid consumption in the field

A molecular approach, using aphid-specific monoclonal antibodies, was used to test the hypothesis that alternative prey can affect predation on aphids by linyphiid spiders. These spiders locate their webs in cereal crops within microsites where prey density is high. Previous work demonstrated that of two subfamilies of Linyphiidae, one, the Linyphiinae, is web-dependent and makes its webs at sites where they were more likely to intercept flying insects plus those (principally aphids) falling from the crop above. The other, the Erigoninae, is less web-dependent, making its webs at ground level at sites with higher densities of ground-living detritivores, especially Collembola. The guts of the spiders were analysed to detect aphid proteins using enzyme-linked immunosorbent assay (ELISA). Female spiders were consuming more aphid than males of both subfamilies and female Linyphiinae were, as predicted, eating more aphid than female Erigoninae. Rates of predation on aphids by Linyphiinae were related to aphid density and were not affected by the availability of alternative prey. However, predation by the Erigoninae on aphids was significantly affected by Collembola density. Itinerant Linyphiinae, caught away from their webs, contained the same concentration of aphid in their guts as web-owners. However, nonweb-owning Erigoninae, living away from Collembola aggregations at web-sites, contained significantly higher concentrations of aphid. For both subfamilies there was evidence of a disproportionate increase in predation on aphids once Collembola populations had declined. It was concluded that nonaphid prey, by helping to maintain spiders in the field, can significantly affect predation on aphids.     

Early sexual dimorphism in the developing gut microbiome of northern elephant seals

The gut microbiome is an integral part of a species' ecology, but we know little about how host characteristics impact its development in wild populations. Here, we explored the role of such intrinsic factors in shaping the gut microbiome of northern elephant seals (Mirounga angustirostris) during a critical developmental window of 6 weeks after weaning, when the pups stay ashore without feeding. We found substantial sex differences in the early‐life gut microbiome, even though males and females could not yet be distinguished morphologically. Sex and age both explained around 15% of the variation in gut microbial beta diversity, while microbial communities sampled from the same individual showed high levels of similarity across time, explaining another 40% of the variation. Only a small proportion of the variation in beta diversity was explained by health status, assessed by full blood counts, but clinically healthy individuals had a greater microbial alpha diversity than their clinically abnormal peers. Across the post‐weaning period, the northern elephant seal gut microbiome was highly dynamic. We found evidence for several colonization and extinction events as well as a decline in Bacteroides and an increase in Prevotella, a pattern that has previously been associated with the transition from nursing to solid food. Lastly, we show that genetic relatedness was correlated with gut microbiome similarity in males but not females, again reflecting early sex differences. Our study represents a naturally diet‐controlled and longitudinal investigation of how intrinsic factors shape the early gut microbiome in a species with extreme sex differences in morphology and life history.     

Electrophoretic analysis of the prey spectrum of Teretrius nigrescens (Lewis) (Col., Histeridae), a predator of Prostephanus truncatus (Horn) (Col., Bostrichidae), in Mexico, Honduras, and Benin

Abstract: Extensive collections of Teretrius nigrescens in Mexico, Honduras, and Benin, were electrophoretically analysed to elucidate the prey spectrum of the predator. Both polyacrylamide gel electrophoresis and cellulose acetate electrophoresis were used. Beetles were sampled with pheromone traps, using the synthetic aggregation pheromone of Prostephanus truncatus , and directly from farmers' maize stores. The proportion of electrophoretically detected prey protein from adult T. nigrescens in pheromone traps was low: of the 1108 specimens analysed, only in 34 cases, prey protein could be clearly identified. More frequently, prey protein was detected in adult T. nigrescens sampled in maize stores in Mexico, Honduras, and Benin, with 87 samples showing distinct prey bands of the total 1214 predators analysed. Of the 241 T. nigrescens larvae sampled in maize stores in Benin, 136 showed distinct bands of prey protein. In all samples, P. truncatus was the most frequently detected prey species. The second most often identified prey species was Sitophilus zeamais . The results are discussed with regard to various methods for prey spectrum analysis and specifically the biology of T. nigrescens .     

Development of molecular diagnostic markers for sharpshooters Homalodisca coagulata and Homalodisca liturata for use in predator gut content examinations

To aid in identifying key predators of Proconiini sharpshooter species present in California, we developed and tested molecular diagnostic markers for the glassy‐winged sharpshooter, Homalodisca coagulata (Say), and smoke‐tree sharpshooter, Homalodisca liturata (Ball) (Homoptera: Cicadellidae). Two different types of markers were compared, those targeting single‐copy sequence characterized amplified regions (SCAR) and mitochondrial markers targeting the multicopy cytochrome oxidase subunit genes I (COI) and II (COII). A total of six markers were developed, two SCAR and four mitochondrial COI or COII markers. Specificity assays demonstrated that SCAR marker HcF5/HcR7 was H. coagulata specific and HcF6/HcR9 was H. coagulata/H. liturata specific. COI (HcCOI‐F/R) and COII (HcCOII‐F4/R4) markers were H. coagulata specific, COII (G/S‐COII‐F/R) marker was H. coagulata/H. liturata specific, and lastly, COII marker (Hl‐COII‐F/R) was H. liturata specific. Sensitivity assays using genomic DNA showed the COI marker to be the most sensitive marker with a detection limit of 6 pg of DNA. This marker was 66‐fold more sensitive than marker Hl‐COII‐F/R that showed a detection limit of 400 pg of DNA. In addition, the COI marker was 4.2‐fold more sensitive than the COII marker. In predator gut assays, the COI and COII markers demonstrated significantly higher detection efficiency than the SCAR markers. Furthermore, the COI marker demonstrated slightly higher detection efficiency over the COII marker. Lastly, we describe the inclusion of an internal control (28S amplification) for predation studies performing predator gut analyses utilizing the polymerase chain reaction (PCR). This control was critical in order to monitor reactions for PCR failures, PCR inhibitors, and for the presence of DNA.     

A comparative analysis of spider prey spectra analyzed through the next‐generation sequencing of individual and mixed DNA samples

As one of the most abundant predators of insects in terrestrial ecosystems, spiders have long received much attention from agricultural scientists and ecologists. Do spiders have a certain controlling effect on the main insect pests of concern in farmland ecosystems? Answering this question requires us to fully understand the prey spectrum of spiders. Next‐generation sequencing (NGS) has been successfully employed to analyze spider prey spectra. However, the high sequencing costs make it difficult to analyze the prey spectrum of various spider species with large samples in a given farmland ecosystem. We performed a comparative analysis of the prey spectra of Ovia alboannulata (Araneae, Lycosidae) using NGS with individual and mixed DNA samples to demonstrate which treatment was better for determining the spider prey spectra in the field. We collected spider individuals from tea plantations, and two treatments were then carried out: (1) The DNA was extracted from the spiders individually and then sequenced separately (DESISS) and (2) the DNA was extracted from the spiders individually and then mixed and sequenced (DESIMS). The results showed that the number of prey families obtained by the DESISS treatment was approximately twice that obtained by the DESIMS treatment. Therefore, the DESIMS treatment greatly underestimated the prey composition of the spiders, although its sequencing costs were obviously lower. However, the relative abundance of prey sequences detected in the two treatments was slightly different only at the family level. Therefore, we concluded that if our purpose were to obtain the most accurate prey spectrum of the spiders, the DESISS treatment would be the best choice. However, if our purpose were to obtain only the relative abundance of prey sequences of the spiders, the DESIMS treatment would also be an option. The present study provides an important reference for choosing applicable methods to analyze the prey spectra and food web compositions of animal in ecosystems.     

Dietary complexity and hidden costs of prey switching in a generalist top predator

1. Variation in predator diet is a critical aspect of food web stability, health, and population dynamics of predator/ prey communities. Quantifying diet, particularly among cryptic species, is extremely challenging, however, and differentiation between demographic subsets of populations is often overlooked.
2. We used prey remains and data taken postmortem from otter Lutra lutra to determine the extent to which dietary variation in a top predator was associated with biotic, spatial, and temporal factors.
3. Biotic data (e.g., sex, weight, and length) and stomach contents were taken from 610 otters found dead across England and Wales between 1994 and 2010. Prey remains were identified to species where possible, using published keys and reference materials. Multi‐model inference followed by model prediction was applied to test for and visualize the nature of associations.
4. Evidence for widespread decline in the consumption of eels (Anguilla anguilla) reflected known eel population declines. An association between eel consumption and otter body condition suggested negative consequences for otter nutrition. Consumption of Cottus gobio and stickleback spp. increased, but was unlikely to compensate (there was no association with body condition). More otters with empty stomachs were found over time. Otter sex, body length, and age‐class were important biotic predictors of the prey species found, and season, region, and distance from the coast were important abiotic predictors.
5. Our study is unique in its multivariate nature, broad spatial scale, and long‐term dataset. Inclusion of biotic data allowed us to reveal important differences in costs and benefits of different prey types, and differences between demographic subsets of the population, overlaid on spatial and temporal variation. Such complexities in otter diet are likely to be paralleled in other predators, and detailed characterization of diet should not be overlooked in efforts to conserve wild populations.

     

Phenology and Genome Size Variation in Allium L. ‐ a Tight Correlation?

Abstract: A very wide diversity of genome size and phenology has been shown in 75 Allium species belonging to all of the six subgenera. The 2C DNA amounts per genome, which range from 16.93 to 63.57 pg, do not show any significant negative correlation with flowering period or any direct relation with foliage leaf dormancy. This means that there is no general correlation between evolution of genome size and life strategies in the differentiation of the genus Allium. This conclusion is rather unexpected in being contrary to the ecological perspective of the nucleotype theory in the genus Allium.     

Strain-level fitness in the gut microbiome is an emergent property of glycans and a single metabolite

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High mortality rates in a juvenile free‐ranging marine predator and links to dive and forage ability

1. High juvenile mortality rates are typical of many long‐lived marine vertebrate predators. Insufficient development in dive and forage ability is considered a key driver of this. However, direct links to survival outcome are sparse, particularly in free‐ranging marine animals that may not return to land.
2. In this study, we conduct exploratory investigations toward early mortality in juvenile southern elephant seals Mirounga leonina. Twenty postweaning pups were equipped with (a) a new‐generation satellite relay data tag, capable of remotely transmitting fine‐scale behavioral movements from accelerometers, and (b) a location transmitting only tag (so that mortality events could be distinguished from device failures). Individuals were followed during their first trip at sea (until mortality or return to land). Two analyses were conducted. First, the behavioral movements and encountered environmental conditions of nonsurviving pups were individually compared to temporally concurrent observations from grouped survivors. Second, common causes of mortality were investigated using Cox's proportional hazard regression and penalized shrinkage techniques.
3. Nine individuals died (two females and seven males) and 11 survived (eight females and three males). All but one individual died before the return phase of their first trip at sea, and all but one were negatively buoyant. Causes of death were variable, although common factors included increased horizontal travel speeds and distances, decreased development in dive and forage ability, and habitat type visited (lower sea surface temperatures and decreased total [eddy] kinetic energy).
4. For long‐lived marine vertebrate predators, such as the southern elephant seal, the first few months of life following independence represent a critical period, when small deviations in behavior from the norm appear sufficient to increase mortality risk. Survival rates may subsequently be particularly vulnerable to changes in climate and environment, which will have concomitant consequences on the demography and dynamics of populations.

     

Epizootiology of a nuclear polyhedrosis virus in European spruce sawfly (Gilpinia hercyniae): the rate of passage of infective virus through the gut of birds during cage tests.

The passage of a nuclear polyhedrosis virus (NPV) of the sawfly, Gilpinia hercyniae, through avian gut was studied during cage tests on Sturnus vulgaris (three individuals), Parus ater (one), Parus caerulus (five), and Parus major (one). Following brief infection feeds, polyhedral inclusion bodies of the virus could be detected in bird feces within 0.5 hr. Peak passage of polyhedra occurred in less than 1 hr and none were detected after 2.5 hr. The feces of all birds remained infective (in bioassay tests using first instar G. hercyniae larvae) to the end of the day of infection while those of nine birds remained infective to the next day and of six birds to the third day. One bird, P. major, was also infective on Days 4, 6, and 7. The infectivity of NPV in feces stored for 2 years at +3°C declined by half. Though the scale of their epizootiological contribution is unknown, the comparatively long retention and passage of infective virus suggests birds may be effective in short- and long-distance transport of baculoviruses.     

Effects of mixed prey on the development and demographic attributes of a generalist predator,Coccinella septempunctata (Coleoptera: Coccinellidae)

Coccinella septempunctata Linnaeus, 1758 (Coleoptera: Coccinellidae) is an aphidophagous ladybird beetle species with cosmopolitan distribution. However, it may also thrive on arthropods other than aphids, when the latter are not readily available. Certain life history traits and demographic attributes of C. septempunctata were determined using seven different diets with different alternations and proportions of an aphid, Lipaphis erysimi (Kaltenbach,1843) and a mite, Tetranychus urticae Koch, 1836, in the laboratory (at 25±0.5°C, 65±5% RH and a photoperiod of 16 h L:8 h D).The larval duration was longest (12.61±6.17 days) when fed an alternating diet of two days aphid/two days mite. Adult longevity was longest (78.65±1.09 days) when fed aphids only and shortest (3.17±1.25 days) when fed mites only. Life expectancy (e_x) was highest (111 days) when adults were fed only aphids and lowest (11.3 days) when fed only mites. Survival rates of ladybird beetles were positively related to an increasing ratio of aphids in their diet. Their ability to prey on both aphids and mites indicates the effectiveness of C. septempunctata as a biological control agent on plants infested with these two pests.