Positive effects of some gyrase inhibitors on survival and development of Pimpla turionellae (Hymenoptera: Ichneumonidae) larvae reared on an artificial diet

Effects of novobiocin, nalidixic, and oxolinic acids on the survival and development of the hymenopterous endoparasitoid Pimpla turionellae L. were investigated alone and in various combinations by rearing larvae aseptically on chemically defined synthetic diets. Effects varied with the levels and kinds of the antibiotics in the tested combinations. Among the tested antibiotics alone, only the lowest level of novobiocin enhanced the postlarval survival. Most of the combinations significantly increased the number of survivors in the postlarval developmental stages. Novobiocin in combination with nalidixic acid (6.0:2.0 mg) in the artificial diet produced a yield of 80.8% pupae and 69.4% adults, respectively. This combination also shortened the developmental time to adult emergence. Similarly, on most of the diets with combinations of the antibiotics, the development of the larvae up to adult stage was noticeably faster than those on the control diets. An approximately equals 50% reduction in the percentages of pupae and adults was recorded by 6.0 mg of novobiocin in combination with 0.75 mg of oxolinic acid compared with the control diet. Although this combination prolonged the complete development of the insect according to other novobiocin and oxolinic acid combinations, developmental time was significantly shorter than that with control diet.     

亚洲玉米螟幼虫对氮、糖的吸收和利用

本文以不同含氮和含糖量的系列人工饲料为材料,研究了亚洲玉米螟Ostrinia fur nacalis(Guen6e)幼虫对氮和糖的吸收和利用。结果表明：幼虫对饲料中氮的吸收能力随虫龄增加而下降,对糖的吸收能力则随之而增强;5龄幼虫对糖的吸收能力随所食饲料中氮含量的增加而增强,而对氮的吸收则与饲料糖含量关系不大,5龄幼虫对氮和糖的消耗速率均随饲料含糖量增加而加快,但幼虫的氮利用率和虫体含氮量则随之下降;幼虫的氮消耗速率随饲料含氮量的增加而增加,而氮利用率则下降,5龄幼虫的氮利用率比3龄幼虫的氮利用率低,导致虫体含氮量的迅速下降。这些结果说明饲料中氮含量影响幼虫对糖的吸收,高龄幼虫对糖的需要量增加而对氮的需要则较稳定,饲料中糖比氮更重要。     

Host rearing is a bottleneck for classical biological control of the cherry bark tortrix: A comparative analysis of artificial diets

This paper describes a comparative analysis of the suitability of three artificial diets for the development of the cherry bark tortrix (CBT), Enarmonia formosana Scopoli (Lepidoptera: Tortricidae), to simplify the rearing process for this species and its potential classical biological control agents. The three diets tested included (1) a pinto bean-based diet modified specifically for the CBT, (2) the diet for codling moth, Cydia pomonella Linnaeus (Lepidoptera: Tortricidae), and (3) the Singh general-purpose diet. Survival from first instar to the pupal stage was very low on the pinto bean, codling moth, and Singh general-purpose diets (29, 0, and 0%, respectively). Survival was consistently greater, yet still low, for larvae that were reared through the first instar on bark and subsequently transferred to the codling moth or Singh general-purpose diets (5 and 32%, respectively). In comparison, larvae started on the pinto bean diet as second instars had a survival rate of 90%, only slightly below that of sibling larvae from the cherry bark control group (100%). Larval development time was fastest on cherry bark (36±2?days), differing significantly from that on the pinto bean diet (started as first instars: 58±2?days; started as second instars: 46±2?days), but not from the development time of larvae on the Singh general-purpose diet (44±3?days). Pupal weights were greatest for specimens from the Singh general-purpose diet (14.9±0.5?mg) and lowest for those from the pinto bean diet (started as first instar: 12.3±0.6?mg; started as second instar: 12.1±0.4?mg). Pupal weights from cherry bark were intermediate (13.5±0.6?mg). Early mortality, resulting primarily from rejection of the diet, remains to be the critical impediment in CBT rearing. It is therefore suggested that a phagostimulant from cherry bark be identified and included in an artificial diet shown to be nutritionally suitable, such as the Singh general-purpose diet or the pinto bean diet.     

In vitro rearing of Toxoneuron nigriceps (Hymenoptera: Braconidae), a larval endoparasitoid of Heliothis virescens (Lepidoptera: Noctuidae) from early second instar to third instar larvae

The early second instar larvae of Toxoneuron nigriceps, a larval endoparastioid of Heliothis virescens, were incubated in artificial rearing media, supplemented with hemolymph of the unparasitized and parasitized fifth instar larvae of the host, H. virescens. The parasitoid larvae were incubated in both a semisolid and liquid form of the artificial rearing medium, and their growth and development were evaluated. The growth in size (increase in length and width), development (molting), and survival of the incubated larvae were observed for 10 days. The incubated larvae exhibited some level of growth in all nine types of media tested, including the control (without host hemolymph). However, ingesting the semisolid rearing media supplemented with the hemolymph from the late fifth instar (day 5, 7 and 9) parasitized host resulted in 100% of the larvae molting to third instars. Some of the in vitro reared third instar larvae demonstrated behavioral changes that could be interpreted as the preparation for cocoon formation or pupation i.e. oral secretion of a whitish material and lots of twisting and turning; however, none produced a cocoon nor pupa.     

二化螟人工饲料研究初报

在温度为(29±1)℃,相对湿度为80%~90%的条件下,分别以3种人工饲料和天然饲料(新鲜水稻茎杆)对二化螟Chilo suppressalisWalker进行连续继代饲养。结果表明,3种人工饲料饲养的二化螟幼虫的生长历期、化蛹率、蛹重、蛹期及羽化率等均与天然饲料的基本接近,而且这3种人工饲料配制简便、成本较低,也不易霉变,是3种较为理想的人工饲料。通过第2代和第3代的继代繁殖表明,3种人工饲料和天然饲料饲养的二化螟的发育情况都稍有下降,但3种人工饲料与天然饲料间不存在显著差异。     

Effects of diet on development of Podisus nigrispinus (Dallas) (Het., Pentatomidae), a predator of the cotton leafworm

Abstract:  The effects of diet on development of Podisus nigrispinus (Dallas) (Het., Pentatomidae) were studied at 25°C, relative humidity of 60 ± 10%, and photoperiod of 14 h light : 10 h dark. Development times of P. nigrispinus nymphs were similar when fed with third or fifth instar larvae of cotton leafworm ( Alabama argillacea Hübner) (Lep., Noctuidae) or Tenebrio molitor L. (Col., Tenebrionidae). When fed with housefly larvae ( Musca domestica L.) (Dipt., Muscidae) or artificial diet, the predator had a longer development time. Independent of diet, instar or sex, the females of P. nigrispinus showed a longer longevity than the males. The total survival of the nymphal stage, on the different diets, varied from 22.46 (fed with housefly) to 77.33% (fed with T. molitor larvae). P. nigrispinus males were heavier when fed with third or fifth instar cotton leafworm larvae than when fed with artificial diet. The weight of the females varied from 37.91 (with artificial diet) to 64.68 mg (with fifth instar cotton leafworm larvae). Independently of the diet, newly emerged females of P. nigrispinus were heavier than the males. Females of P. nigrispinus which were fed with fifth instar cotton leafworm larvae had heavier ovaries than those fed other diets.     

Utilization of fatty acids by Pieris brassicae reared on artificial and natural diets

The fatty acid composition of Pieris brassicae was measured from larvae reared on four different diets. Pieris can alter the composition of fatty acids in the diet through selective incorporation and synthesis. Oleate is preferentially accumulated on artificial diets (15·9 per cent in diet, 43·8 per cent in neutral lipid (NL) of fifth instar larvae), but not equally on natural diets (18·1 per cent in Brassica napus, 25·6 per cent in the NL of fifth instar larvae). Incorporation of linolenate appears to depend on the concentration of both linolenate and linoleate in the diet. With dietary levels of 35·7% linolenate and 32·2% linoleate, fifth instar larvae contain 12·2 and 16·0 per cent, respectively, of these acids. With 45·8% linolenate and 12·5% linoleate in the diet, fifth instar larvae contain 44·1 and 11·6 per cent of these acids, respectively, in the NL. Palmitoleate is actively synthetized on the artificial diets; with trace amounts of dietary palmitoleate, fifth instar larvae have 9·3 per cent of this acid in the NL. Pieris regulates the uptake of linoleate from the diet at the intestinal wall as was shown by linoleic acid-1-¹⁴C, and is unable to convert dietary linoleate to any of the 18-carbon analogues. The female apparently accumulates linolenate into egg phospholipids on the artificial diet, but in general the fatty acid composition of the eggs resembles that of the fat body.     

Biological and immune response of Galleria mellonella (Lepidoptera: Pyralidae) to sodium tetraborate

Inorganic insecticides are commonly used in urban pest management because of their low mammalian toxicity. We tested the effects of sodium tetraborate (ST) on life parameters of greater wax moth, Galleria mellonella (L.) (Lepidoptera: Pyralidae), to determine its sublethal toxicity on the insect. Survival, development, adult longevity, and fecundity of the wax moth were investigated by rearing larvae on artificial diets containing ST at concentrations of 0.005, 0.1, 0.2, or 0.3%. Larvae reared on medium at the highest concentration of ST (0.3%) had significantly decreased survival to the seventh instar and prolonged time required to reach the seventh instar. This concentration reduced pupa and adult yields to 12.5%, and it also prolonged development by 5 d. ST did not significantly influence adult longevity. Dietary ST led to significant decreases in fecundity and egg viability. Oviposition of survivors at the highest ST concentration (0.3%) was completely inhibited. Lysozyme content was decreased in larval hemolymph and fat body at high dietary ST concentrations. Fat body lysozyme content was significantly increased two-fold for larvae reared on diet at the lowest concentration of ST (0.005%). However, the highest concentration (0.3%) dramatically decreased fat body lysozyme content from 0.12 +/- 0.013 to 0.006 +/- 0.003 mg/ml in seventh instars. We infer that sublethal levels of dietary ST substantially influence life history parameters and immunocompetence in G. mellonella.     

Comparison of the growth and development of beet armyworm,Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) on two different diets

Beet armyworm (Spodoptera exigua (Hübner)) (Lepidoptera: Noctuidae) is the most economically important sugar beet (Beta vulgaris) pest worldwide. In this study, a comparison was made between two different diets: one was based on Merkx diet (Holidic diet) and the other was based on sugar beet leaf (Oligidic diet). Results showed that the whole development time from larvae to adult between two diets (Merkx and leaf) was not significantly different. For example, developmental time from first instar larvae to adult in Merkx diet was 11.33?days, whilst developmental time of larvae to adult when larvae fed with sugar beet leaf was 10.33?days. However, analysis of variance showed that in some cases like development time of the first instar, third instar and fifth instar larvae and pupae was significantly different between two treatments (p?<?0.05). Larval weight showed differences when larvae fed on Merkx diet and sugar beet leaves. For example, significant differences were shown between first, third and fourth instar larvae weight when larvae fed on Merkx diet and sugar beet leaves (p?<?0.05). However, significant differences were not observed between weight of second and fifth instar as well as pupae weight when larvae fed on Merkx diet and sugar beet leaves (p?>?0.05).     

Performance of Lymantria xylina (Lepidoptera: Lymantriidae) on artificial and host plant diets

Lymantria xylina Swinhoe (Lepidoptera: Lymantriidae) is a serious defoliator of hardwood and fruit trees in Taiwan. The larvae of L. xylina feed on >63 species of host plants, belonging to 29 families. Because a large number of larvae are needed for the production of nucleopolyhedrosis virus (NPV) or other related studies, the development of a suitable artificial diet is very important for the mass rearing of this moth in the laboratory. In this study, eight artificial diets, modified from different formulas, and one host plant, Liquidambar formosana Hance, were used to feed L. xylina caterpillars. Through various bioassays (first instar survival trial and long- and short-term feeding trials), the most suitable diet for the L. xylina was selected by performance comparisons with L. formosana. After the first instar survival trial, two of the diets were discarded, because no larva survived on these diets. The results of the long-term feeding trial indicated that the larvae grew successfully on only three kinds of artificial diet. Finally, results of the short-term feeding trial revealed that a diet (diet A), modified from the gypsy moth, Lymantria dispar (L.), formula diet, was the most appropriate for the L. xylina. Larvae fed on diet A had better survival rate, pupal weight, adult size, efficiency of conversion, and relative growth rate than larvae fed on other diets; they did not grow as well as those fed on L. formosana, however, except for pupal and adult weight, and approximate digestibility. In summary, diet A was found to be the best of the artificial diets for the L. xylina and is suitable for mass rearing of this moth in the laboratory.     

The relative importance of temperature and diet to larval development and adult size of the winter stonefly, Soyedina carolinensis (Piecoptera: Nemouridae)

SUMMARY. 1. Soyedina carolinensis Claassen, a leaf shredding stonefly, was reared in a series of three laboratory experiments from early instar to adult on different species of deciduous leaves and at various constant and fluctuating temperature regimes.
2. Experiment 1, which involved rearing larvae on fourteen different leaf diets at ambient stream temperatures, showed that diet significantly affected larval growth and adult size but did not affect overall developmental time.
3. Experiment 2, which involved rearing larvae on five different leaf diets at each of three fluctuating temperature regimes (viz ambient White Clay Creek (WCC), ambient WCC+3°C, and ambient WCC+6°C), showed that: (i) adding 6°C to the normal temperature regime of WCC was lethal to 99% of the larvae regardless of diet; and (ii) warming WCC by 3°C did not affect developmental time but did significantly reduce adult size relative to adults reared at WCC temperatures on certain diets.
4. Experiment 3, which involved rearing larvae on five different leaf diets at each of five constant temperatures (viz 5, 10, 15, 20, 25°C), showed that: (i) temperature significantly affected the mortality, growth, and development time of larvae whereas diet only affected larval growth and mortality; (ii) temperatures at or near 10°C yielded maximum larval growth and survival for most diets; (iii) at 5°C, larval mortality was high and growth was low resulting in a few small adults for most diets; (iv) larval mortality was at or near 100% at 15°C regardless of diet; and (v) no larvae survived at 20 and 25°C.     

Insect cytokine growth-blocking peptide may regulate density-dependent phase trait of cuticular melanization in the larval armyworm,Mythimna separata

Growth-blocking peptide (GBP) is a 25 amino acid insect cytokine found in lepidopteran insects that has diverse biological activities such as larval growth regulation, paralysis induction, cell proliferation and stimulation of immune cells. Density-dependent phase polyphenism is a phenomenon of phenotypic plasticity in which the expression of a variety of traits can be affected by local population density. In the present study, the armyworm Mythimna separata larvae with four rearing densities (1 larva/vial, 2 larvae/vial, 4 larvae/vial and 6 larvae/vial) were tested for cuticular melanization and body weight throughout the third-fifth instar, and the functional role of GBP in regulating the changes was investigated. The results indicated that when reared at high densities, the larvae exhibited less body weight and more degree of cuticular melanization than larvae reared at low densities. The gene expression of GBP in armyworm larvae showed an initial rise and then decline trend with increased rearing densities in the third to fifth instar. Compared with control, more degree of cuticular melanization was observed in GBP-injected larvae (500 ng/larva in volume 50 μL) than that in Ringer’s solution-injected counterparts. Furthermore, the gene expression level of dopa decarboxylase and prophenoloxidase increased significantly in GBP-injected fifth instar larvae from 6 h to 12 h after injection, suggesting the role of GBP in modulating density-dependent phase trait of armyworm cuticular melanization.     

Effects of B group vitamins and choline chloride on the development and protein synthesis in the male larvae of Pimpla turionellae L. (Hym., Ichneumonidae)

The effects on the development and protein levels in the larvae of male Pimpla turionellae L. of different amounts of B group vitamins namely thiamin, riboflavin, Ca-pantothenate, nicotinic acid, pyridoxine-HCl, folic acid and biotin and a growth factor choline chloride in the meridic diet were investigated. Among the vitamins tested, choline chloride and biotin were found to have quantitatively critical values in the diet and increased amounts of nicotinic acid and folic acid individually in the control diet showed positive effects on development and protein levels. A decrease in the levels of Ca-pantothenate and pridoxine-HCl caused decreases in protein levels, whereas variations in the levels of thamin and riboflavin did not have any significant effect.     

Rearing <Emphasis Type="Italic">Chrysoperla externa</Emphasis> Larvae on Artificial Diets

We tested three artificial diets for rearing larvae of Chrysoperla externa (Hagen) (Neuroptera: Chrysopidae), aiming at reducing the production costs of this predator. Two of the diets come from studies with other species of lacewings, and the third is a modification described in this paper. All diets were based on animal protein and were supplied to 2nd and 3rd instar larvae, whereas 1st instar larvae received eggs of Anagasta kuehniella (Zeller) (Lepidoptera: Pyralidae). We evaluated the preimaginal duration and survival, adult size, longevity and fecundity, egg hatchability, and predatory capacity of larvae produced. The performance of the diets was followed for seven generations. The diet we describe showed to be the best among the artificial diets tested. Our results show that C. externa can be successfully reared on artificial diets during second and third instars, reducing in 90% the dependency on eggs of A. kuehniella.     

Influence of plant,animal and mixed resources on development of the zoophytophagous plant bug Campylomma verbasci (Hemiptera: Miridae)

The mullein bug, Campylomma verbasci (Meyer-Dür) (Hemiptera: Miridae), exploits both plant and animal resources. The aim of this study was (1) to evaluate the quality of different plant, animal and mixed diets (i.e. plant material or prey from the overwintering host, the summer host or the laboratory rearing) for the development and survival of mullein bug, and (2) to evaluate if the suitability of the resource changes according to the nymphal instar of C. verbasci. Mullein bug nymphs were reared individually in Petri dishes containing different diets and observed daily until reaching adulthood or dying. The rearing diet (living potato aphids?+?potato leaf?+?Ephestia eggs?+?apple pollen) was a high-quality diet. The medium-quality diets were apple fruit?+?apple pollen, Ephestia eggs and Sitotroga eggs. The low-quality diets included mullein leaf, frozen aphids, living aphids?+?potato leaf and apple fruit?+?living aphids?+?potato leaf. Finally, the inappropriate diets (when no nymphs reached adulthood) were agar gel, apple fruit, apple pollen, apple leaf, potato leaf, frozen spider mites and frozen conspecifics. When comparing the value of the diets for different instars, living aphids?+?potato leaf was a high-quality diet for third instar. Apple pollen and frozen conspecifics were low-quality diets for third instar. ‘Red Delicious’ fruit was a high-quality diet for fifth instar and low-quality for third instar. ‘Honeycrisp’ fruit constituted an inappropriate diet whatever the instar. Thus, a whole mixed diet maximises development and survival of C. verbasci nymphs.     

Artificial diets for rearing the coconut hispine beetle Brontispa longissima and its parasitoid Asecodes hispinarum

We tested artificial diets for rearing the coconut hispine beetle, Brontispa longissima, a serious invasive pest of coconut (Cocos nucifera) in Southeast Asia. We examined three artificial diets that were identical except for their agar content. The survival rate from hatching to adult emergence was 26.0% when beetles were reared on a ‘soft diet’ (20 g/l agar), 16.0% on a ‘hard diet’ (40 g/l agar), and 41.0% on a ‘mixed diet’, in which the hard diet was used for the first instar and the soft diet for later instars. Females raised as larvae on the mixed diet and then as adults on the soft diet laid few eggs, which did not hatch. However, females reared on the mixed diet as larvae and then on fresh leaves as adults consistently laid eggs that hatched. We then examined the suitability of B. longissima larvae reared on the mixed diet as hosts for the larval parasitoid Asecodes hispinarum, a specialist parasitoid of this beetle. Of the oviposited hosts, 75.0% became mummified and 41.7% produced adult wasps. These results indicate that the mixed diet may be useful for rearing B. longissima larvae as hosts for the rearing of A. hispinarum.     

The effects of boric acid-induced oxidative stress on antioxidant enzymes and survivorship in Galleria mellonella

Larvae of the wax moth, Galleria mellonella (L.), were reared from first instar on a diet supplemented with 156, 620, 1,250, or 2,500 ppm boric acid (BA). The content of malondialdehyde (MDA, an oxidative stress indicator), and activities of the antioxidant enzymes [superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST), and glutathione peroxidase (GPx)] were determined in the fat body and hemolymph in the 7th instar larvae and newly emerged pupae. Relative to control larvae, MDA was significantly increased in larval hemolymph, larval and pupal fat body, but decreased in the pupal hemolymph. Insects reared on diets with 156- and 620-ppm BA doses yielded increased SOD activity but 1,250- and 2,500-ppm doses resulted in decreased SOD activity in larval hemolymph. SOD activity was significantly increased but CAT was decreased in the larval fat body. High dietary BA treatments led to significantly decreased GST activity. However, they increased GPx activity in larval hemolymph. Dietary BA also affected larval survival. The 1,250- and 2,500-ppm concentrations led to significantly increased larval and pupal mortality and prolonged development. In contrast, the lowest BA concentration increased longevity and shortened development. We infer that BA toxicity is related, at least in part, to oxidative stress management.     

Effect of the bacterial DNA gyrase inhibitors, novobiocin, nalidixic acid, and oxolinic acid, on oxidative phosphorylation

When incubated with isolated intact rat liver mitochondria, novobiocin and nalidixic acid act as uncouplers of oxidative phosphorylation; they stimulate oxygen uptake and inhibit ATP synthesis. Novobiocin is about as powerful an uncoupler as is 2,4-dinitrophenol, nalidixic acid is somewhat less powerful, and oxolinic acid exerts no inhibition whatsoever at the concentrations used. The three inhibitors are without effect on oxidative phosphorylation in Escherichia coli nor does novobiocin affect this process in a novobiocin-permeable mutant of yeast. While it would appear that oxolinic acid may be a relatively specific tool for the manipulation of the superhelicity of DNA in complex systems such as mammalian mitochondria and intact mammalian cells, the specificity of each of these inhibitors may depend upon the particular conditions and species used and such experiments require adequate controls on oxidative phosphorylation.     

Facultative monophagy as a consequence of prior feeding experience: behavioral and physiological specialization in Colias philodice larvae

Summary Although newly-emerged Colias philodice readily accept Medicago sativa, Melilotus alba, and Coronilla varia, fifth instar larvae reared on any single plant species display a highly significant induced feeding preference for their rearing host. Forced host-switching reveals that fifth instar relative growth rate (RGR) on M. sativa and M. alba is significantly reduced by prior feeding on either alternative host. Moreover, regardless of rearing diet, switching to a novel host during the fifth instar results in reduced RGR, relative consumption rate (RCR), efficiency of conversion of digested food (ECD), and pupal weight. These results support the hypothesis that induction of feeding preference is an adaptive response that predisposes larvae to feed on the plant species they are most capable of utilizing for growth.     

A method for rearing and assessing the health of the sugarcane pest Dermolepida albohirtum (Coleoptera: Scarabaeidae)

Rearing insects in controlled conditions is a prerequisite to supply high-quality specimens for bioassays. However, while artificial diets and standardized rearing methods have been developed for many phytophagous insects, especially Lepidoptera, there are limited published diets for root-feeding Coleoptera which are commonly fed either grass roots or pieces of vegetables as a simple alternative to artificial diets during bioassays. These feeding options, while convenient, can be considered suboptimal as they do not maximise the insects' development and health. Additionally, it is also important to develop standardised screening methods designed to test sublethal effects of control agents which may have repellent, antifeedant, antimetabolic and/or delayed mortality effects. The greyback canegrub (Dermolepida albohirtum, Waterhouse) is the most damaging native pest of Australian sugarcane, but no rearing method or artificial diet has ever been developed for this species. Our objectives were to improve bioassay methodology for D. albohirtum by describing and developing standard rearing and health assessment protocols. We describe a successful rearing method to raise healthy D. albohirtum larvae with a total of 48.8% of first instars successfully moulting to the second instar. We also tested a modified artificial diet which increased the weight, size and food uptake of larvae compared to traditional methods (i.e., pieces of carrots). For example, the average weight increase of larvae fed with the modified diet was 3.4 times higher than for carrot-fed larvae while modified diet-fed larvae were 2.1 times wider than if they were fed with carrots. Finally, we developed a method to measure larval activity which can be used to identify sublethal effects of control agents such as effects on activity level. Our methods may also be applied to improve bioassay methodology for other root-feeding Coleoptera.