Does nutrition-related stress carry over to spruce budworm,Choristoneura fumiferana (Lepidoptera: Tortricidae) progeny?

Three different patterns of feeding of sixth-instar spruce budworm, Choristoneura fumiferana Clemens were simulated in the laboratory. Larvae were fed artificial diets whose nitrogen and total soluble sugar contents varied according to levels similar to those found in three types of balsam fir, Abies balsamea (L.) Miller foliage (current-year foliage from middle and lower crown sections and one-year-old foliage). The biological performance of offspring was studied according to the nutrition of their parents. Although food quality had no impact on pupal weight of female parents and individual mean egg weight, progeny fitness was influenced by parental nutrition. Old foliage simulated diet, poor in nitrogen, clearly affected the early larval development of progeny, especially the percent of egg hatch and first-instar survival. Lower crown current-year foliage simulated diet, with low total soluble sugar content, reduced the first-instar survival of the progeny. However, the selective pressure exerted by low food qualities on the parental generation and on the early stages of their progenies resulted in C. fumiferana populations having higher tolerance to starvation and higher survival after the diapause period. These results highlighted the potentially direct and indirect effects of C. fumiferana parental nutrition on the next generation. The patterns of feeding of parental generations would appear to affect the quality and size of subsequent populations through several selections on the different life-history stages of both generations.     

Parental effects in Pieris rapae in response to variation in food quality: adaptive plasticity across generations?

1. Herbivores using seasonal resources must cope with variation in the quality of their host plants. The effects of variation in protein concentration of artificial diet and glucosinolate concentration in canola, Brassica napus, on Pieris rapae parental and progeny growth were investigated. 2. The hypothesis that parents respond to variation in food quality by altering the phenotype of their progeny to enhance progeny fitness was tested. Consistent with previous studies, P. rapae was not affected strongly by variation in the protein concentration of artificial diet and had equal mass on completing development. 3. The mass of individual eggs of P. rapae progeny was correlated negatively with the amount of protein in the diet on which parents fed. Moreover, mothers reared in extreme conditions (high and low protein) produced progeny that grew best under those conditions. These potentially adaptive parental effects were detected early in progeny growth but not later in their development. 4. Early larval growth of P. rapae was affected negatively by increasing glucosinolates in B. napus plants, although no effects of glucosinolates were detected later in growth or on the progeny's phenotype. 5. Thus, evidence is presented that variation in food quality (protein concentration) has major consequences for the progeny of P. rapae. Given the multivoltine life history of P. rapae and the seasonal differences in food quality it encounters, such parental effects may be adaptive.     

Carry over effects of the entomopathogen Bacillus thuringiensis ssp. Kurstaki on Choristoneura fumiferana (Lepidoptera: Tortricidae) progeny under various stressful environmental conditions

Abstract

• 1 In the present study, we documented the lethal and sublethal effects of the entomopathogen Btk on spruce budworm and its progeny under various environmental conditions. We hypothesized that aerial spray of Bacillus thuringiensis ssp. kurstaki (Btk) could affect the biological performance of the surviving spruce budworm (Choristoneura fumiferana Clem.) populations and their progenies and that Btk sublethal effects could be widened by other types of stress (i.e. temperature conditions and changes in food suitability from year to year).
• 2 The results from a 3‐year field experiment indicated that Btk treatments decreased the fitness of the surviving larvae whatever the prevailing temperature and nutritional conditions.
• 3 The detrimental Btk effects on the parental generation carried over to the offspring. The percent of egg hatch and first‐instar survival were negatively affected by Btk whatever other stress spruce budworm parents underwent.
• 4 The present study also highlighted the fact that the effects of temperature and nutritional stress suffered by the parents could carry over to the next generation. Balsam fir flowering, which provided larvae with pollen rich in nitrogen, favoured both the parental generation and the fitness of their offspring. Spruce budworm mothers allocated to their progenies large amounts of energy reserves (triglycerides and glycogen) that greatly enhanced the survival of the early stages.
• 5 Egg hatch and the survival of first‐instar larval progeny were drastically affected when their parents had reduced larval growth as a result of exposure to cool temperatures that had desynchronized insect and bud phenology.
• 6 Budworm mothers submitted to negative impacts of previous defoliation allocated low amounts of energy reserves to their progeny. This lack of energy associated with unfavourable temperature conditions (i.e. high temperatures in late summer and in early fall and an extended cool period in spring) drastically reduced survival of diapausing second‐instar larvae.
• 7 These results highlight the importance of considering the various sources of stress when attempting to evaluate the impact of a control agent on an insect pest population and its progenies.

     

Phase-dependent locomotor activity in first-stadium nymphs of the desert locust, Schistocerca gregaria: effects of parental and progeny rearing density

This study examined the effects of parental and progeny rearing density on locomotor activity of 1st-stadium nymphs of the desert locust, Schistocerca gregaria, using an actograph. Progeny obtained from solitarious (isolated-reared) or gregarious (crowd-reared) locusts were reared in isolation or in a group of 30 nymphs. Crowding after hatching had a slight influence on mean activity shortly after the start of measurements, but no clear effect was detected until day 2, when maximum activity during the 6-24 h of observation was significantly higher than that of the nymphs kept in isolation. On the other hand, the effects of parental rearing density on locomotor activity manifested at all ages examined (0-2 days old). Progeny of gregarious locusts showed consistently higher activity than those of solitarious locusts. In newly hatched nymphs, the effect of parental rearing density was explained by variation in body size at hatching, one of the phase-dependent characteristics. Hatchling body color was also correlated with locomotor activity and body weight. Similar levels of locomotor activity were exhibited when green, solitarious and black, gregarious nymphs were similar in body weight. These results suggested that parental rearing density indirectly influences locomotor activity in the progeny shortly after hatching by affecting their body size as eggs or hatchlings.     

Transgenerational effects of parental larval diet on offspring development time, adult body size and pathogen resistance in Drosophila melanogaster

Environmental conditions experienced by parents are increasingly recognized to affect offspring performance. We set out to investigate the effect of parental larval diet on offspring development time, adult body size and adult resistance to the bacterium Serratia marcescens in Drosophila melanogaster. Flies for the parental generation were raised on either poor or standard diet and then mated in the four possible sex-by-parental diet crosses. Females that were raised on poor food produced larger offspring than females that were raised on standard food. Furthermore, male progeny sired by fathers that were raised on poor food were larger than male progeny sired by males raised on standard food. Development times were shortest for offspring whose one parent (mother or the father) was raised on standard and the other parent on poor food and longest for offspring whose parents both were raised on poor food. No evidence for transgenerational effects of parental diet on offspring disease resistance was found. Although paternal effects have been previously demonstrated in D. melanogaster, no earlier studies have investigated male-mediated transgenerational effects of diet in this species. The results highlight the importance of not only considering the relative contribution each parental sex has on progeny performance but also the combined effects that the two sexes may have on offspring performance.     

Diapause incidence of progeny in relation to parental geographic origin,host plant and rearing density in the cabbage beetle,Colaphellus bowringi

The cabbage beetle, Colaphellus bowringi Baly (Coleoptera: Chrysomelidae), which is widely distributed in China, undergoes an imaginal diapause in the soil. In this study, we investigated the influences of parental geographic origin, host plant, and rearing density on the diapause incidence of progeny in this beetle. In studies conducted at 25 °C and L12:D12 photoperiod, the proportions of adults entering diapause were significantly different among latitudes from which the parents had been collected. The incidence of diapause increased with increasing latitude. Reciprocal cross tests between post‐diapause adults from different latitudes showed that there were significant differences in diapause incidence between pure‐bred and hybrid adults, suggesting that diapause incidence among progeny was determined by both the female and male, although the female appeared to have a stronger effect than the male. These results revealed that parental origin has an important influence on progeny diapause. Parental host plant species not only affected diapause incidence of the current generation, but also affected the incidence of diapause in the progeny produced by the non‐diapause parents; yet, the parental host plant had no influence on the incidence of diapause in the progeny produced by the post‐diapause parents. Rearing density of the parental generation had no significant effect on the incidence of diapause in the progeny.     

Life history and cost analysis for continuous rearing of Perillus bioculatus (Heteroptera: Pentatomidae) on a zoophytogenous artificial diet

The impact of a zoophytogenous, insect-free artificial diet on the developmental rate, life history parameters, and fertility was examined over 11 consecutive generations for domesticated Perillus biocullatus (F.) (Heteroptera: Pentatomidae). This study showed that when fed an insect-free artificial diet during both the nymphal and adult stages, developmental times were prolonged, and the net reproductive rates (R0) and the intrinsic rates of increase (r(m)) were significantly lower than when fed larval Trichoplusia ni at both nymphal and adult stages. Moreover, the cost to rear P. bioculatus on the artificial diet approached 1.1 times the cost of rearing P. bioculatus on natural prey. These results demonstrate the effectiveness and potential cost-savings of the zoophytogenous artificial diet for rearing a beneficial pentatomid.     

The price of eggs: increased investment in egg production reduces the offspring rearing capacity of parents

Understanding the selective pressures shaping the number of offspring per breeding event is a key area in the study of life-history strategies. However, in species with parental care, costs incurred in offspring production, rather than rearing, have been largely ignored in both theoretical and empirical studies until relatively recently. Furthermore, the few experimental studies that have manipulated production costs have not yet teased apart effects that operate via the parental phenotype from effects on the quality of the resulting young. To examine whether increased egg production influences parental brood rearing capacity independently of effects operating via egg quality, we experimentally increased egg production in gulls and then examined their capacity to rear a control clutch. We found that the capacity of parents to rear the control brood was substantially reduced solely as a consequence of having themselves produced one extra egg. The paradox that, in many species, parents apparently aim for fewer young per breeding event than the experimentally and theoretically demonstrated optimum, has partly arisen from the failure to take into account the constraints imposed by production costs.     

Parental Self-Feeding Effects on Parental Care Levels and Time Allocation in Palestine Sunbirds

The trade-off between parents feeding themselves and their young is an important life history problem that can be considered in terms of optimal behavioral strategies. Recent studies on birds have tested how parents allocate the food between themselves and their young. Until now the effect of food consumption by parent birds on their food delivery to their young as well as other parental activities has rarely been studied. I have previously shown that parent Palestine sunbirds (Nectarinia osea) will consume nectar and liquidized arthropods from artificial feeders. However, they will only feed their young with whole arthropods. This provided a unique opportunity to experimentally manipulate the food eaten by parents independent of that fed to their offspring. Here, I hypothesized that parents invest in their current young according to the quality of food that they themselves consume. Breeding pairs with two or three nestlings were provided with feeders containing water (control), sucrose solution (0.75 mol) or liquidized mealworms mixed with sucrose solution (0.75 mol). As food quality in feeders increased (from water up to liquidized mealworms mixed with sucrose solution): 1) Parents (especially females) increased their food delivery of whole arthropod prey to their young. 2) Only males increased their nest guarding effort. Nestling food intake and growth rate increased with increasing food quality of parents and decreasing brood size. These results imply that increasing the nutrient content of foods consumed by parent sunbirds allow them to increase the rate at which other foods are delivered to their young and to increase the time spent on other parental care activities.     

Effect of host diet on acceptance of eastern spruce budworm eggs byTrichogramma minutum

The acceptance behaviour of the parasitic wasp, Trichogramma minutum Riley (Hymenoptera: Trichogrammatidae), was studied on two types of egg masses of eastern spruce budworm (SBW), Choristoneura fumiferana Clemens. Host eggs were obtained from moths that had been fed balsam fir or artificial diet during their larval stages. Parasitoids were reared from both types of hosts. Host acceptance assays were conducted where females were provided with a single egg mass and with a choice between egg masses. Regardless of the parasitoid's rearing host, females parasitised hosts reared from artificial diet more often than those from balsam fir. Response was consistent in both no-choice and choice assays. Rejection of hosts tended to occur during the examining stage of behaviour, suggesting that acceptability is judged externally. If a host was accepted, the same number of eggs were laid per egg mass regardless of treatment.     

Larval experience induces adult aversion to rearing host plants: a novel behaviour contrary to Hopkins' host selection principle

1. Hopkins' host selection principle (HSP) states that insects should prefer foliage from their rearing host plant over that of an alternative host. 2. The current study tested whether eastern spruce budworm, Choristoneura fumiferana (Clem.) (Lepidoptera: Tortricidae), that were laid and developed on, respectively, resistant and susceptible white spruce Picea glauca (Moench) Voss, showed differences in their feeding and oviposition preferences for these two hosts. 3. The data revealed that previous experience of spruce budworm on a host tree type does not influence the host acceptance and feeding behaviour of later larval stages. However, adult budworm reared on resistant white spruce needles preferentially selected susceptible white spruce needles as the host for their progeny, whereas those reared on susceptible needles showed no preference. 4. To the authors' knowledge, this is the first report of an insect showing an oviposition preference for the non‐rearing host plant. This would tend to increase mixing between insects from susceptible and resistant trees. The present results thus argue against Hopkins' HSP and suggest that learned aversion to resistant foliage experienced by larvae is carried into the adult stage.     

Life table parameters, reproductive rate, intrinsic rate of increase, and estimated cost of rearing Podisus maculiventris (Heteroptera: Pentatomidae) on an artificial diet.

The impact of an insect-free artificial diet provided at nymphal and/or adult stage upon the developmental rate, life table parameters, and fertility table parameters was examined for Podisus maculiventris (Say). This study showed that when fed an insect-free artificial diet during both the nymphal and adult stage, developmental time was prolonged, preoviposition period was extended, and reproductive rate (R0) and intrinsic rate of increase (r) were significantly lower than when fed larval insect prey at both nymphal and adult stages. Additionally, feeding larval prey to adults reared as nymphs on an artificial diet significantly increased the proportion of fertile females, the number of eggs laid by mated females, the reproductive rate and intrinsic rate of increase, but the mean generation time was not significantly different. Likewise, feeding artificial diet to adults reared on larval prey resulted in a significant reduction in reproductive rate and intrinsic rate of increase. The "realized" cost to rear P. maculiventris on the artificial diet was calculated (as the cost to double the population size) using raw material cost, fertility table parameters and doubling time values. Raw material cost for rearing P. maculiventris colony on Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae) was only 1.4 times higher than the cost of artificial diet raw materials required to rear the same size colony. However, the realized cost of rearing was 3.5 times higher when rearing on artificial diet because of the prolonged developmental time and reduced reproductive output. The cost efficiency of rearing a beneficial insect on an artificial diet that decreases the intrinsic rate of increase of a colony is discussed, as well as the advantages and disadvantages of supplementing adult diets with natural prey at the reproductive stage.     

沿海拔梯度的表型可塑性影响银胶菊叶甲的取食效率和发育(英文)

【目的】本研究旨在弄清楚生态气候因子是否影响尼泊尔银胶菊Parthenium hysterophorus盛产区中银胶菊叶甲Zygogramma bicolorata成虫后代的形态学测量参数和基础生物学。【方法】 从尼泊尔Kathmandu (24℃, 海拔1 400 m, 温带气候), Chitwan (25℃, 海拔415 m, 热带或亚热带气候)和Mahendranagar (34℃, 海拔229 m, 湿润 亚热带气候)地区采集银胶菊叶甲成虫,在最适室内条件下评估各生态气候区中其F₁代4龄幼虫和雌成虫的取食特征。我们假定在自由取食和最适非生物条件下饲养的后代将不会受到食物和气温的间接影响,不管亲本属于哪一生态气候区,后代利用食物的能力一致。【结果】然而,我们发现,尽管在最适条件下饲养,银胶菊叶甲后代的体型和取食特征与其亲本相似。体型大的亲本(来自Kathmandu地区),其后代体型也大,但是其食物利用效率比体型小的亲本(来自Chitwan 和Mahendranagar地区)所产后代小。这3个生态气候区中,与幼虫相比正在发育的雌成虫均表现出体重增加而食物利用效率降低。【结论】这些发现说明,食物和最适非生物条件的间接影响可能不影响银胶菊叶甲后代的表型可塑性,后代的体型和取食特征与亲本相似。发生的可遗传的变化可能是由于种内的遗传多样性。我们期望这些发现有助于理解气候变化背景下银胶菊叶甲成虫的表型可塑性、分布格局和取食行为。     

Suitability of pines and other conifers as hosts for the invasive Mediterranean pine engraver (Coleoptera: Scolytidae) in North America

The invasive Mediterranean pine engraver, Orthotomicus erosus (Wollaston) (Coleoptera: Scolytidae), was detected in North America in 2004, and it is currently distributed in the southern Central Valley of California. It originates from the Mediterranean region, the Middle East, and Asia, and it reproduces on pines (Pinus spp.). To identify potentially vulnerable native and adventive hosts in North America, no-choice host range tests were conducted in the laboratory on 22 conifer species. The beetle reproduced on four pines from its native Eurasian range--Aleppo, Canary Island, Italian stone, and Scots pines; 11 native North American pines--eastern white, grey, jack, Jeffrey, loblolly, Monterey, ponderosa, red, Sierra lodgepole, singleleaf pinyon, and sugar pines; and four native nonpines--Douglas-fir, black and white spruce, and tamarack. Among nonpines, fewer progeny developed and they were of smaller size on Douglas-fir and tamarack, but sex ratios of progeny were nearly 1:1 on all hosts. Last, beetles did not develop on white fir, incense cedar, and coast redwood. With loblolly pine, the first new adults emerged 42 d after parental females were introduced into host logs at temperatures of 20-33 degrees C and 523.5 or 334.7 accumulated degree-days based on lower development thresholds of 13.6 or 18 degrees C, respectively.     

韭蛆人工饲料配方筛选及饲养效果比较

【目的】研究实验室条件下韭蛆 Bradysia odoriphaga Yang et Zhang大量饲养的方法以解决试虫不足的问题。【方法】本实验在25℃,相对湿度75%,光周期16L:8D条件下,通过控制变量法筛选出人工饲料配方,运用两性生命表软件,分析比较人工饲料与韭菜Allium tuberosum的饲喂效果。【结果】对不同成分和不同含量的8种韭蛆人工饲料配方筛选,结果确定最优的人工饲料配方成分和含量为：平菇粉6 g,韭菜粉5 g,琼脂粉1.25 g,山梨酸0.04 g,苯甲酸0.05 g,酵母粉0.5 g,维生素C 2.5 g和水50 mL。该配方和韭菜饲养效果相当,不存在显著差异（P>0.05）。同时,两性生命表结果表明,人工饲料饲喂的韭蛆平均世代周期23.6800 d,韭菜饲喂的韭蛆世代周期为25.9000 d,两者之间差异显著（P<0.05）。生殖生物学特性分析发现：韭菜和人工饲料饲喂的韭蛆雌成虫净生殖率分别为16.3200和41.1800/d,内禀增长率分别为0.1078和0.1570/d。【结论】筛选的人工饲料和韭菜均可以稳定地完成韭蛆的世代繁殖,且人工饲料饲喂效果略优于韭菜,另外人工饲料较韭菜不但具有低成本省人工的优点,而且可解决韭菜反季节供应和农药残留问题,保证稳定的虫源供给。     

Ecophysiological adaptations of black spruce (<Emphasis Type="Italic"> Picea mariana</Emphasis>) and tamarack (<Emphasis Type="Italic"> Larix laricina</Emphasis>) seedlings to flooding

Black spruce [ Picea mariana (Mill.) B.S.P.] and tamarack [ Larix laricina (Du Roi) K. Koch] are the predominant tree species in boreal peatlands. The effects of 34 days of flooding on morphological and physiological responses were investigated in the greenhouse for black spruce and tamarack seedlings in their second growing season (18 months old). Flooding resulted in reduced root hydraulic conductance, net assimilation rate and stomatal conductance and increased needle electrolyte leakage in both species. Flooded tamarack seedlings maintained a higher net assimilation rate and stomatal conductance compared to flooded black spruce. Flooded tamarack seedlings were also able to maintain higher root hydraulic conductance compared to flooded black spruce seedlings at a comparable time period of flooding. Root respiration declined in both species under flooding. Sugar concentration increased in shoots while decreasing in roots in both species under flooding. Needles of flooded black spruce appeared necrotic and electrolyte leakage increased over time with flooding and remained significantly higher than in flooded tamarack seedlings. No visible damage symptoms were observed in flooded tamarack seedlings. Flooded tamarack seedlings developed adventitious roots beginning 16 days after the start of flooding treatment. Adventitious roots exhibited significantly higher root hydraulic conductivity than similarly sized flooded tamarack roots. Flooded black spruce lacked any such morphological adaptation. These results suggest that tamarack is better able to adjust both morphologically and physiologically to prolonged soil flooding than black spruce seedlings.     

Effects of exposing Drosophila melanogaster parents to ethanol on expression of vestigial in their progeny

When parental Drosophila melanogaster were chronically exposed at 28 degrees C or 24 degrees C to ethanol during their larval and pupal stages of development, their progeny, produced when parents were 5-16-day-old adults, showed modified expression of vestigial alleles in heterozygous and homozygous combinations. Parental alcohol effects were dependent on parental rearing temperature. We conclude that parental environment (alcohol, temperature) causes heritable but transitory changes in progeny phenotype that are elicited by exposure of germ cells to alcohol.     

人工饲料与天然饲料饲养条件下烟青虫的生长发育和繁殖力比较

报道饲养烟青虫HelicoverpaassultaGue澖ee的一种人工饲料,比较了烟青虫取食该饲料和天然饲料的主要生物学指标,测定了6龄幼虫对2种食料的转化和利用率。该人工饲料的主要成分是麦胚、黄豆粉、酵母粉和烟叶粉,天然饲料为烟草叶片。结果表明:取食人工饲料的烟青虫幼虫生长发育快,化的蛹较大,成虫寿命长,产卵量大,所产卵孵化率高;取食人工饲料的烟青虫与取食烟草蒴果的相比,前者相对取食量和近似消化率较高,相对生长率无显著性差异。说明烟青虫对该人工饲料有较好的适应性,可以作为繁殖烟青虫的实用饲料。     

Interspecific variation in resistance of two host tree species to spruce budworm

Woody plants regularly sustain biomass losses to herbivorous insects. Consequently, they have developed various resistance mechanisms to cope with insect attack. However, these mechanisms of defense and how they are affected by resource availability are not well understood. The present study aimed at evaluating and comparing the natural resistance (antibiosis and tolerance) of balsam fir (Abies balsamea [L.] Mill.) and white spruce (Picea glauca [Moench) Voss] to spruce budworm, Choristoneura fumiferana (Clem.), and how drainage site quality as a component of resource availability affects the expression of resistance over time (6 years). Our results showed that there are differences in natural resistance between the two tree species to spruce budworm, but it was not significantly affected by drainage quality. Balsam fir exhibited higher foliar toxic secondary compounds concentrations than white spruce in all drainage classes, resulting in lower male pupal mass, survival and longer male developmental time. This, however, did not prevent spruce budworm from consuming more foliage in balsam fir than in white spruce. This response suggests that either natural levels of measured secondary compounds do not provide sufficient toxicity to reduce defoliation, or spruce budworm has developed compensatory mechanisms, which allow it to utilize food resources more efficiently or minimize the toxic effects that are produced by its host's defensive compounds. Larvae exhibited lower pupal mass and higher mortality in rapidly drained and subhygric sites. Drainage class also affected the amount of foliage destroyed but its impact varied over the years and was probably influenced by climatic variables. These results demonstrate the complexity of predicting the effect of resource availability on tree defenses, especially when other confounding environmental factors can affect tree resource allocation and utilization.     

Factors affecting oral regurgitation by larval spruce budworm

This study evaluated factors that influence the regurgitation behaviour of sixth instar spruce budworm, Choristoneura fumiferana Clemens (Lepidoptera: Tortricidae), reared on balsam fir, Abies balsamea (L.) Mill. (Pinaceae), under various experimental conditions in the laboratory. Upon physical disturbance, larvae discharged a median volume of regurgitant of 0.4 μl when fed and 1.6 μl when food‐deprived. Larvae deprived of food for 24 or 48 h disgorged more regurgitant than larvae feeding on balsam fir foliage, and the effect was consistent for laboratory‐reared and field‐collected larvae. The water content of the foliage fed upon by larvae had no immediate impact on the volume of regurgitant; following a 24‐h period of food deprivation, however, larvae that previously fed on fresh foliage discharged >2.5 times more regurgitant than larvae that previously fed on dry foliage. Self‐regulated regurgitation by larvae, measured using the number of regurgitant stains on filter paper, was >10 times higher when larvae had access to balsam fir foliage than when they were starved. The number of larvae confined inside the Petri dish (one or four individuals) had a relatively small effect on regurgitation. Larvae were deterred from feeding when balsam fir needles were entirely covered with regurgitant, but not when only a portion of the foliage was treated. These results suggest that the regurgitant does not serve as resource marking or spacing pheromone. The high level of regurgitation by larvae after contact with ants suggests that the regurgitant has evolved in part as a defence mechanism against natural enemies.