Rapid cold hardening in the olive fruit fly Bactrocera oleae under laboratory and field conditions

A rapid cold hardening response was studied in females and males of the olive fruit fly Bactrocera (Dacus) oleae. When laboratory-reared females and males were transferred and maintained from the rearing temperature of 24 °C for 2 h to –6.5 °C approximately 5% survived. However, conditioning of both females and males for 2 h at various temperatures from 0 to 10 °C before their exposure for 2 h to –6.5 °C increased survival to 80 to 92%. A similar rapid cold hardening response in both females and males was also induced through gradual cooling of the flies at a rate of approximately 0.4 °C per min. The rapid increase in cold tolerance after prior conditioning of the flies to low temperatures, was rapidly lost when they returned to a higher temperature of 24 °C. In the field, in late February and early March, females and males were capable of a rapid cold hardening response. After exposure to the critical temperature they suffered a high mortality when tested in the afternoon and low mortality early in the morning on consecutive days, probably because of differences in the prevailing field temperatures a few hours before testing. This plasticity of cold tolerance gained through rapid cold hardening may allow the flies to survive during periods of the year with great fluctuation in circadian temperatures.     

Oviposition response and development of the egg-pupal parasitoid Fopius arisanus on Bactrocera oleae, a tephritid fruit fly pest of olive in the Mediterranean basin

To date, information is wanting with regard to the use of new exotic parasitoids against olive fruit fly, Bactrocera (=Dacus) oleae (Gmelin) (Diptera: Tephritidae), a serious pest of olives Olea europaea L., in the Mediterranean basin. We investigated the oviposition response and developmental biology on B. oleae of Fopius (=Biosteres) arisanus (Sonan) (=Opius oophilus Fullaway) (Hymenoptera: Braconidae), an egg-pupal parasitoid of tephritid fruit flies, never tested before as a potential parasitoid of this host. Our results showed that olive fruits infested with B. oleae eggs exerted a relevant attraction to gravid F. arisanus and represented a stimulus for oviposition. Nevertheless they were not as attractive to female parasitoids as the Mediterranean fruit fly, Ceratitis capitata Wiedemann (Diptera: Tephritidae), eggs infested papaya fruits (Carica papaya L.). In our experimental conditions, F. arisanus completed development in B. oleae within 33 ± 1.7 days (males) and 35 ± 1.6 (females). Increases in host egg to female parasitoid ratios of 1:1, 5:1, 10:1 and 20:1 corresponded with decreases in the percentage of B. oleae parasitisation and host killing but corresponded also with increases in absolute parasitisation. Our findings are discussed in light of possibilities of utilising F. arisanus for biological control of olive fruit fly.     

The activation of amino acid analogues by phenylalanyl- and tyrosyl-tRNA synthetases from plants

Partially purified preparations of Phe- and Tyr-tRNA synthetases were obtained from seed or seedlings of Phaseolus aureus, Delonix regia and Caesalpinia tinctoria, and the ability of a variety of structural analogues of Phe or Tyr to act as alternative substrates or inhibitors was tested. 3-Hydroxymethylphenylalanine, a natural product of C. tinctoria, formed a particularly effective substrate for the Tyr-tRNA synthetase from P. aureus. The structural features commensurate with substrate activity in an analogue molecule are discussed.     

Multiple mating in Anastrepha fraterculus females and its relationship with fecundity and fertility

The occurrence of female remating has been widely reported in insects and the frequency at which it occurs and the factors driving females’ remating behavior have been shown to be both species specific and variable within species. Herein, we studied the remating behavior of females from a well established laboratory colony and a wild population of the South American fruit fly, Anastrepha fraterculus (Wiedemann) (Diptera: Tephritidae), under laboratory conditions. Latency to first mating (number of days from the onset of the experiment until the first copula) was shorter for remating females than for females that did not remate. Two‐day fecundity was higher for females that did remate than for monogamous females. Egg hatch was sustained after remating and was not affected by the number of times the female mated. However, when females willing to remate were prevented from doing so, percent egg hatch showed a significant drop. These results and the fact that remating occurred more often in more fecund females than in less fecund ones suggest that remating may be a response to sperm depletion. Remating frequency was similar in laboratory and wild flies, but 2‐day fecundity was higher for laboratory than for wild females of similar mating status. Also, the length of the refractory period (time between first and second copulation) was longer for wild than for laboratory females. Differences between strains could be the result of artificial selection. Results are discussed from a theoretical and applied perspective in the context of direct benefits to females.     

Comparative field studies of various traps and attractants of the olive fruit fly,Bactrocera oleae

In the framework of an ongoing testing and refining process of a mass trapping method, using a combination of semiochemicals, for the control of the olive fruit fly,Bactrocera (Dacus) oleae, four trap designs, three trap colors, six different food attractants, and two pheromone formulations were compared under field conditions. No differences were observed between the trap designs and the food attractants tested. Trap color had a significant effect in preference tests only. Traps combining food attractants and pheromones attracted higher numbers of both male and female flies but the differences became statistically significant only in cases of low trap densities. Pheromones enclosed in cyclodextrenes were not as attractive as standard pheromone formulations. The findings of these tests allow a choice among trap types and semiochemicals for cost reduction and convenience, but not for enhancement of the efficacy of the method.     

Evaluation of entomopathogenic nematodes against the olive fruit fly, Bactrocera oleae (Diptera: Tephritidae)

Infectivity of six entomopathogenic nematode (EPNs) species against Bactrocera oleae was compared. Similar infection levels were observed when third-instar larvae were exposed to infective juveniles (IJs) on a sand-potting soil substrate. When IJs were sprayed over naturally infested fallen olives, many larvae died within treated olives as well as in the soil; Steinernema feltiae caused the highest overall mortality of 67.9%. In addition, three laboratory experiments were conducted to optimize a time period for S. feltiae field application. (1) Abundance of fly larvae inside fallen olives was estimated over the 2006–2007 season with the highest number of susceptible larvae (3 mm and larger) per 100 olives being observed during December, 2006. (2) S. feltiae efficacy against fly larvae dropped to the soil post-IJ-application was determined. B. oleae added to the substrate before and after nematode application were infected at similar levels. (3) Effect of three temperature regimes (min–max: 10–27, 6–18, and 3–12 °C) corresponding to October through December in Davis, California on S. feltiae survival and infectivity was determined. After 8 weeks, the IJs at the 3–12 °C treatment showed the highest survival rate. However, the cold temperature significantly limited S. feltiae infectivity. Our results demonstrate that B. oleae mature larvae are susceptible to EPN infection both in the soil and within infested olives. Being the most effective species, S. feltiae may have the potential to suppress overwintering populations of B. oleae. We suggest that November is the optimal time for S. feltiae field application in Northern California.     

Relative flight responses of Rhagoletis indifferens as influenced by crowding, sex, and resources

Although flight is believed to be the primary mechanism for dispersal in the Western cherry fruit fly, Rhagoletis indifferens Curran (Diptera: Tephritidae), an orchard pest of both sweet (Prunus avium L.) and sour (Prunus cerasus L.) (Rosaceae) cherry crops, the movement of these flies between host patches is difficult to quantify in the field. A tethered flight mill system was used in the laboratory to examine the flight behaviour of sexually mature flies exposed to different levels of conspecific contact and resource availability. A complete 2 × 2 × 3 factorial design compared the relative influence of the factors ‘context’ (crowded, isolated), ‘sex’ (female, male), and ‘resources’ (low = food only; medium = food + leaf; high = food + leaf + cherries) on flight performance measures including distance flown, net trial time, and stopping patterns. Rather than using a minimum time or distance to determine trial length, flight observations were continued for each fly until a behavioural protocol based on stopping time was met. In this protocol each successful trial was composed of three consecutive flight intervals and included a minimum of three stops lasting a combined total of 5 min. Of the 160 flies tested, 86.9% flew <500 m on the flight mill. Individuals from both sexes were capable of maximum flights in the same order of magnitude, ca. 3 km on the flight mill. Distance flown was significantly influenced by ‘context’ such that crowded individuals flew >1.5-fold farther than isolated individuals. Sex influenced the frequency and duration of stops made, with females stopping more often and longer than males. Although females and males in high resource treatments had the shortest net trial times, the factor ‘resources’ did not produce any highly significant main effects, but did generate significant interaction terms with the factors ‘context’ and ‘sex’, suggesting that past experience with ‘resources’ modifies individual flight behaviour. We have shown for the first time using a tethered flight mill system that R. indifferens flight behaviour is context dependent and sensitive to adult crowding. The implications of this study for improved field experiments on dispersal are discussed.     

The accumulation and metabolism of amino acids by Biomphalaria glabrata, a freshwater pulmonate snail

Freshwater pulmonate snails (Biomphalaria glabrata), pre-treated under bacteriostatic conditions, were incubated in 10 ml of standard medium containing various U-¹⁴C-labelled amino acids at concentrations of 10 μM. Measurements of mass-specific accumulation rates (MSARs) based on HPLC and the accumulation of U-¹⁴C-labelled amino acids into snail tissues have shown unequivocally for the first time that freshwater snails achieved a net accumulation of all the amino acids tested, including aminoisobutyrate (AIB), aspartate, alanine and a mixture of 13 amino acids. There were no significant differences between the MSAR values determined by HPLC from those based on the use of radiolabelled amino acids, whereas MSAR values for control snails were negligible and significantly less. Incubation of snails in media containing radiolabelled aspartate and a mixture of amino acids showed that the accumulated amino acids were readily distributed through the snail’s tissues and then metabolized. The ecological and biochemical questions arising from the fact that freshwater snails are capable of net accumulation of exogenous amino acids at naturally occurring concentrations and subsequent metabolic conversion, contrary to widely held views, are addressed.     

湖南道地白芷氨基酸含量与组成分析

白芷是我国常用中药材的一种,关于白芷镇痛的有效成分,主要认为是异欧前胡素、欧前胡素等香豆素类具有解痉、镇痛等作用的挥发油类化学成分,但白芷镇痛的成分及其机理尚不十分清楚,是否有其他物质也具有一定的药理作用也不得而知,比如说某些药效性氨基酸。该研究采用自动氨基酸分析仪测定了湖南道地茶陵白芷的蛋白质类氨基酸和游离氨基酸的含量,并分析其氨基酸组成。结果表明:总氨基酸检出除Asn以外的其他16种蛋白质氨基酸,含量最高的Arg占总氨基酸的31.21%,接近1/3;必需氨基酸总量达27.01%,其中含量最高的是Leu,占总必需氨基酸的24.14%;药效氨基酸比例较高,在酸水解产物总氨基酸和游离氨基酸中分别达到73.89%和85.78%,其中Arg的含量最高,达1.383g·100g-1,占比为42.24%,游离氨基酸中γ-氨基丁酸、鸟氨酸含量也较高。此外,还含有少量的高半胱氨酸和鹅肌肽等游离非蛋白质氨基酸和短肽;必需氨基酸的组成接近WHO/FAO的建议摄入值,但Met+Cys的RC值最小,为第一限制性氨基酸。这些药效性氨基酸和游离氨基酸可能是茶陵白芷具有良好药效的一个因素,而且茶陵白芷的氨基酸组成和配比较合理、符合人体需要,在强化含硫氨基酸的基础上具有开发成为新型药食同源保健性食品的潜力。研究结果可为进一步研究白芷的药理作用提供依据。     

Thermal responses in the citrus fruit fly,Dacus tsuneonis: evidence for a pupal diapause

Thermal responses controlling pupariation and adult eclosion in a citrus fruit fly,Dacus tsuneonis (Miyake), were studied to understand the winter biology of this species. When mature larvae were exposed to various temperature conditions, the highest percentage of pupariation was obtained at 15 °C, although the variance at this temperature was greater than at 20 °C or 25 °C. Pupariation occurred most rapidly at 20 °C and an alternating temperature with a mean of 15 °C. At constant 15 °C, pupae failed to emerge as adults. Pupae were characterized by a reduced respiration rate, which is typical of a diapausing pupa. When insects were stored at different temperatures for 45 days after pupariation, and then transferred to 25 °C, adult eclosion occurred earlier when the initial temperature was 10 °C than when it was 5 °C or 15 °C. Adult eclosion occurred most synchronously and pupal mortality was lowest when insects were stored at 15 °C for 90 days before incubation at 25 °C. These results strongly suggest thatD. tsuneonis enters a pupal diapause.     

Effect of age and mating on pheromone production in the female olive fruit fly, Dacus oleae (Gmel.)

Virgin female olive fruit flies began producing sex pheromone on the third day after emergence. Production of sex pheromone appears to be cyclical with peaks of production recurring at about 10-day intervals, each peak lasting 2–3 days. As the female ages, the quantity of sex pheromone produced during the period of high pheromone production decreases, but it is always higher than the quantity present in the pheromone gland during the rest of female life.Mated females produced less sex pheromone the first 10 days after mating which then increased to near the same level of that in virgin females.     

Development, survival and fecundity of the urine fly, Scatella (Teichomyza) fusca and predation by the black dumpfly, Hydrotaea aenescens

Development, survival and fecundity for Scatella (Teichomyza) fusca Maquart (Diptera: Ephydridae) were studied at 20 ± 1 °C and 85 ± 10% r.h. Mean (± S.E.) developmental times of eggs, larvae and pupae were 4.0 ± 0.06, 14.9 ± 0.19 and 14.6 ± 0.11 days, respectively, the mean (± S.E.) survival of the original egg cohort to the start of larval, pupal and adult stage being 77.2 ± 3.2%, 54.5 ± 2.6% and 47.9 ± 3.5%, respectively. Females and males displayed approximately straight survivorship curves during adult life, implying constant mortality rates. Mean (± S.E.) adult longevity was 41.6 ± 2.98 days for females and 51.2 ± 3.91 days for males. Assuming a stable age distribution the population consisted of 56% eggs, 31% larvae, 6% pupae and 7% adults. Oviposition peaked when females were 25 days old, and the highest reproductive values (RV_x) (mean ± S.E.) ranged from 129.1 ± 7.57 to 138.5 ± 6.83 for individuals 17–27 days old. A female deposited a mean (± S.E.) of 614.7 ± 35.9 female eggs over a maximum life span of 93 days. The basic reproductive rate (R ₀) (mean ± S.E.) was 173.0 ± 14.2 female offspring per female and the intrinsic rate of natural increase of female individuals (r) (mean ± S.E.) was 0.088 ± 0.001 day^-1. The mean (± S.E.) generation time (T) was 57.8 ± 0.78 days. In cultures with equal numbers of first instar S. fusca larvae and predacious third instar larvae of Hydrotaea aenescens (synonymous Ophyra aenescens) Wiedemann, mean (± S.E.) survival to the adult stage of S. fusca (16.7 ± 8.8%) was significantly lower than in controls with S. fusca alone (58.3 ± 7.4%). The potential significance of predation by H. aenescens on S. fusca in pig farms is discussed.     

Asymmetry as a predictor of growth, fecundity and survival

Measures of developmental stability such as fluctuating asymmetry have been assumed to predict individual performance because asymmetry reflects an inability to cope with stressful situations, and because asymmetry hampers locomotion. However, the magnitude of this relationship between important fitness components (growth, fecundity, survival) and asymmetry has never been assessed. Based on a literature survey, estimates of the correlation between asymmetry and the three fitness components are presented. Pearson's correlation coefficients weighted for sample size between asymmetry and growth, fecundity, and survival, respectively, were –0.15, –0.35, and –0.25, respectively, with all three coefficients being highly significant. All three relationships were extremely robust given very large fail-safe numbers. The results were independent of whether studies or species were used as units of analysis. Hence asymmetry is a robust predictor of performance in fitness domains such as growth, fecundity, and survival, although only accounting for 2.1%, 12.3%, and 6.0% of the variance. This may be of importance for studies of sexual selection, but also for ecological and conservation biological studies, where the performance of individuals or groups of individuals are assessed.     

Recognition of specific patterns of amino acid inhibition of growth in higher plants, uncomplicated by glutamine-reversible `general amino acid inhibition''

The complexity of the regulatory mechanisms that govern amino acid biosynthesis, particularly in multibranched pathways, frequently results in sensitivity to growth inhibition by exogenous amino acids. Usually the inhibition caused by a given amino acid(s) is relieved by another amino acid(s), thus indicating the cause of inhibition to be a specific interference with endogenous formation of the latter amino acid(s). We recently summarized the evidence that Nicotiana silvestris (and probably most higher plants), in suspension culture, exhibits a separate phenomenon of amino acid mediated growth inhibition called general amino acid inhibition. Every amino acid provokes general amino acid inhibition except for

Full-size image

-glutamine. In fact,

Full-size image

-glutamine completely overcomes general amino acid inhibition. We have now demonstrated that specific amino acid inhibition can be recognized and characterized at the level of growth inhibition without interference caused by general amino acid inhibition by the simple provision of exogenous

Full-size image

-glutamine. Several examples of specific amino acid inhibition of growth were demonstrated in N. silvestris. In one case,

Full-size image

-threonine inhibits growth partially in the presence of

Full-size image

-glutamine. The residual amino acid inhibition was overcome by the additional presence of

Full-size image

-lysine and

Full-size image

-methionine, indicating that exogenous

Full-size image

-threonine specifically inhibits the biosynthesis of both

Full-size image

-lysine and

Full-size image

-methionine. As a second example, the

Full-size image

-valine-mediated inhibition of growth that persisted in the presence of

Full-size image

-glutamine was overcome by

Full-size image

-isoleucine, indicating that exogenous

Full-size image

-valine inhibits

Full-size image

-isoleucine biosynthesis. The use of amino acid analogs as experimental tools for biochemical-genetic studies in higher plants is also complicated by general amino acid inhibition. Conditions were demonstrated under which p-fluorophenylalanine and m-fluorotyrosine could be used as specific antimetabolites of

Full-size image

-phenylalanine and

Full-size image

-tyrosine biosynthesis without interference from general amino acid inhibition. We thus present a rigorous basis for recognition of specific relationships between metabolic branches that can guide detailed enzymological analyses.     

Effect of constant temperatures on development of the walnut husk fly,Rhagoletis completa

The developmental rates of various life stages ofRhagoletis completa Cresson (Diptera: Tephritidae) were determined in the laboratory at seven different constant temperatures: 8, 12, 16, 20, 24, 28, and 32±1°C, RH 80±10%, photoperiod L 16∶D8. Preoviposition developmental rate was fastest at 28°C (10±1 days, mean±SD) and slowest at 12°C (26±1 days). About 83% of the females deposited eggs at 20 and 24°C and only 25% oviposited at 32°C. Females laid the highest number of eggs at 24°C and the lowest at 8°C. Egg development increased with increasing temperatures up to 28°C, then declined. The fastest egg development was noticed at 28°C (55±1 h) and slowest at 8°C (389±2 h). Over 90% egg hatch was observed at temperatures between 12 and 32°C, but decreased to 73% at 8°C. Larval development was fastest also at 28°C (20±0.2 days). Over 65% pupation was recorded at 20 and 24°C, but decreased to 15% at 32°C and 12% at 8°C. Pupal development was most rapid at 24°C (53±1 days) and slowest at 8°C (162±2 days). More than 70% of adult emergence was noticed in treatments between 16 and 24°C but decreased to 20% at 8°C. Based on a linear regression model of temperature-development rate relationship, the lower developmental thresholds were determined to be 6.6, 5.3, 2.9, and 5°C for preoviposition, egg, larval, and pupal stages, respectively. Based on a non-linear developmental rate model, the upper developmental thresholds were 34°C for preoviposition, egg, and larval stages and 30°C for pupal stage.     

Changes in the levels of plant total phenols and free amino acids induced by two cereal aphids and effects on aphid fecundity

Abstract:  Phenols and amino acids have previously been involved in the resistance of cereals to aphids. In the present study, the mean total concentrations of both compounds were quantitatively estimated from leaf-tissue samples collected at three growth stages (third, fifth and seventh leaf stages) from maize and barley seedlings and then correlated with the mean 7-day fecundity of Rhopalosiphum padi (L.) and Sitobion avenae (F.) (Homoptera, Aphididae) reared separately on the corresponding growth stages of each cereal. Results revealed an inverse correlation between the total concentration of each chemical in aphid-infested maize and barley plants and the fecundity of each aphid on every growth stage of the cereals. Moreover, R. padi led to an increase in total amino acids when reared on maize or barley without affecting the phenol concentration, while S. avenae induced a decrease in phenol levels but did not change the total amino acids on each growth stage of the plants. The significance of these results for understanding potential differences in requirements of cereal aphids for host selection and acceptance are discussed.     

Naphthoquinone amino acid derivatives,synthesis and biological activity as proteasome inhibitors

The ubiquitin-proteasome system has been largely investigated for its key role in protein degradation mechanisms that regulate both apoptosis and cell division. Because of their antitumour activity, different classes of proteasome inhibitors have been identified to date. Some of these compounds are currently employed in the clinical treatment of several types of cancer among which multiple myeloma. Here, we describe the design, chemistry, biological activity and modelling studies of a large series of amino acid derivatives linked to a naphthoquinone pharmacophoric group through variable spacers. Some analogues showed interesting inhibitory potency for the β1 and β5 subunits of the proteasome with IC₅₀ values in the sub-µm range.     

The amino acid sequence of hemoglobin II from the symbiont-harboring clamLucina pectinata

The cytoplasmic hemoglobin II from the gill of the clamLucina pectinata consists of 150 amino acid residues, has a calculatedM _m of 17,476, including heme and an acetylated N-terminal residue. It retains the invariant residues Phe 44 at position CD1 and His 65 at the proximal position F8, as well as the highly conserved Trp 15 at position A12 and Pro 38 at position C2. The most likely candidate for the distal residue at position E7, based on the alignment with other globins, is Gln 65. However, optical and EPR spectroscopic studies of the ferri Hb II (Kraus, D. W., Wittenberg, J. B., Lu, J. F., and Peisach, J.,J. Biol. Chem. 265, 16054–16059, 1990) have implicated a tyrosinate oxygen as the distal ligand. Modeling of theLucina Hb II sequence, using the crystal structure of sperm whale aquometmyoglobin, showed that Tyr 30 substituting for the Leu located at position B10 can place its oxygen within 2.8 Å of the water molecule occupying the distal ligand position. This structural alteration is facilitated by the coordinate mutation of the residue at position CD4, from Phe 46 in the sperm whale myoglobin sequence to Leu 47 inLucina Hb II.     

Effects of branched‐chain amino acid supplementation on fecundity,lifespan and flight ability of Helicoverpa armigera

The effects on cotton bollworm, Helicoverpa armigera, of supplementing the benchmark larval feeding formula with branched‐chain amino acids (BCAAs) were analyzed. The results show that supplementary BCAAs can increase adult longevity, and are conducive to improving development and reproductive capacity. In particular, by supplementing the ration with the three proteinogenic BCAAs at concentrations ten times that of the benchmark formula, all the selected physiological indices of cotton bollworm health were improved. The larval duration, pupae weight, pupation rate, fecundity and adult longevity for specimens fed the supplemented formula were 15.8 days, 0.348 g, 100%, 1340.7 eggs and 27.3 days, respectively. The same indicators for larvae fed the benchmark formula were 16.6 days, 0.306 g, 97.9%, 1167.0 eggs and 12.0 days, respectively. This study provides a reference for future research on amino acid supplementation and increasing lifespans