Search

Jaburetox (Jbtx) is an insecticidal peptide derived from Canavalia ensiformis urease, whose mechanism of action is not completely elucidated. We employed behavioral, electromyographical and electrophysiological protocols to identify the cellular and molecular targets involved in the Jbtx entomotoxicity in cockroaches and locusts. In Nauphoeta cinerea, Jbtx (32 μg/g) altered the locomotory behaviour inducing a significative decrease in the distance travelled followed by a significant increase in stopped time (52 ± 85 cm and 2573 ± 89 s, p < .05, n = 40). Jbtx (8 to 32 μg/g body weight, respectively) also increased the leg and antennae grooming activities (p < .05, n = 40, respectively). Jbtx (8 to 16 μg/g) induced a maximum neuromuscular blockade of 80.72% (n = 6, p < .05) and was cardiotoxic, decreasing the cockroach heart rate. The electrophysiological profiles of both muscle and nerve of L. migratoria showed that Jbtx (2.5 × 10 -7 and 2.5 × 10 -3 μg/ body weight) induced a significant increase in the amplitude of nerve action potentials (n = 5, p < .05). Voltage clamp analysis of Jbtx (200 nM) applied in Xenopus laevis oocytes heterologously expressed with Nav 1.1 channels showed a significant increase in the sodium currents. In conclusion, this work revealed that the entomotoxic activity of Jbtx involves complex behavioral alterations that begins with an initial activation of voltage-gated sodium channels., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The Asian Migratory locust (Locusta migratoria migratoria L.) is a pest that continuously threatens crops in the Amudarya River delta near the Aral Sea in Uzbekistan, Central Asia. Its development coincides with the growing period of its main food plant, a tall reed grass (Phragmites australis), which represents the predominant vegetation in the delta and which cover vast areas of the former Aral Sea, which is desiccating since the 1960s. Current locust survey methods and control practices would tremendously benefit from accurate and timely spatially explicit information on the potential locust habitat distribution. To that aim, satellite observation from the MODIS Terra/Aqua satellites and in-situ observations were combined to monitor potential locust habitats according to their corresponding risk of infestations along the growing season. A Random Forest (RF) algorithm was applied for classifying time series of MODIS enhanced vegetation index (EVI) from 2003 to 2014 at an 8-day interval. Based on an independent ground truth data set, classification accuracies of reeds posing a medium or high risk of locust infestation exceeded 89% on average. For the 12-year period covered in this study, an average of 7504 km 2 (28% of the observed area) was flagged as potential locust habitat and 5% represents a permanent high risk of locust infestation. Results are instrumental for predicting potential locust outbreaks and developing well-targeted management plans. The method offers positive perspectives for locust management and treatment of infested sites because it is able to deliver risk maps in near real time, with an accuracy of 80% in April-May which coincides with both locust hatching and the first control surveys. Such maps could help in rapid decision-making regarding control interventions against the initial locust congregations, and thus the efficiency of survey teams and the chemical treatments could be increased, thus potentially reducing environmental pollution while avoiding areas where treatments are most likely to cause environmental degradation., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The morphological, physiological and behavioural differences between solitarious and gregarious desert locusts are so pronounced that one could easily mistake the two phases as belonging to different species, if one has no knowledge of the phenomenon of phenotypic plasticity. A number of phase-specific features are hormonally controlled. Juvenile hormone promotes several solitarious features, the green cuticular colour being the most obvious one. The neuropeptide corazonin elicits the dark cuticular colour that is typical for the gregarious phase, as well as particular gregarious behavioural characteristics. However, it had to be concluded, for multiple reasons, that the endocrine system is not the primary phase-determining system. Our observation that longevity gets imprinted in very early life by crowding of the young hatchlings, and that it cannot be changed thereafter, made us consider the possibility that, perhaps, epigenetic control of gene expression might be, if not the missing, a primary phase-determining mechanism. Imprinting is likely to involve DNA methylation and histone modification. Analysis of a Schistocerca EST database of nervous tissue identified the presence of several candidate genes that may be involved in epigenetic control, including two DNA methyltransferases (Dnmts). Dnmt1 and Dnmt2 are phase-specifically expressed in certain tissues. In the metathoracic ganglion, important in the serotonin pathway for sensing mechanostimulation, their expression is clearly affected by crowding. Our data urge for reconsidering the role of the endocrine system as being sandwiched in between genetics and epigenetics, involving complementary modes of action., (Copyright © 2011 Elsevier Inc. All rights reserved.)

In the last decade, important progress has been made in the experimental analysis of the endocrine mechanisms controlling reproduction and phase transition in locusts. Phase transition is a very fascinating, but complex, phenomenon of phenotypic plasticity that is triggered by changes in population density and can lead to the formation of extremely devastating hopper bands and adult gregarious locust swarms. While some phase characteristics change within hours, others appear more gradually in the next stage(s), or even in the next generation(s). In adults, the phase status also has a major influence on the process of reproduction. A better understanding of how solitarious locusts become gregarious and how this switch affects reproductive physiology may result in novel strategies to fight locust plagues. In this paper, we will review the current knowledge concerning this close interaction between locust phase polyphenism and reproduction.

The interaction between two pathogens, the microsporidian Paranosema locustae Canning and the fungus Metarhizium anisopliae var. acridum Driver and Milner was studied under laboratory conditions in an attempt to develop an improved method of microbial control for the desert locust, Schistocerca gregaria Forskål. Fifth-instar locust nymphs, reared in the laboratory, were treated with various concentrations of one of the two pathogens or with both pathogens. The numbers of locusts killed were recorded each day and the production of pathogen spores within the dead locusts was assessed at the end (day 21) of each experiment. Locust nymphs treated with both P. locustae and M. anisopliae died sooner than nymphs infected with only one of the pathogens. At the lower concentrations of pathogen tested, the effects of the two pathogens were additive. At the higher concentrations the combined effects were synergistic. In terms of locust mortality, there was no evidence of any antagonistic effects between the two pathogens. However, the production of spores by P. locustae was reduced considerably when the host insects were infected also with M. anisopliae. For example, nymphs treated initially with P. locustae and then treated 3 and 10 days later with M. anisopliae produced 3-20 times and 2.5-8 times fewer spores, respectively, than nymphs treated only with P. locustae. Hence, in areas where M. anisopliae is applied, the natural persistence of P. locustae in the local grasshopper and locust populations may be diminished.

The gut bacterial community from four species of feral locusts and grasshoppers was determined by denaturing gradient gel electrophoresis (DGGE) analysis of bacterial 16S rRNA gene fragments. The study revealed an effect of phase polymorphism on gut bacterial diversity in brown locusts from South Africa. A single bacterial phylotype, consistent with Citrobacter sp. dominated the gut microbiota of two sympatric populations of Moroccan and Italian locusts in Spain. There was evidence for Wollbachia sp. in the meadow grasshopper caught locally in the UK. Sequence analysis of DGGE products did not reveal evidence for unculturable bacteria and homologies suggested that bacterial species were principally Gammaproteobacteria from the family Enterobacteriaceae similar to those recorded previously in laboratory reared locusts.

In a manual sequence alignment experiment we accidentally observed that the locust adipokinetic hormone (AKH) precursor-related peptides (APRPs), which are peptides contained in the AKH-preprohormones, display sequence similarity with human growth hormone-releasing factor (GRF). So far, a counterpart for GRF, either structurally or functionally, has not yet been reported in any non-chordate. This could give new insights into the origin of the GRF superfamily, as it would mean that in some invertebrates (locusts), the structural homologues of glucagons (AKHs) and GRFs (APRPs) are located on the same precursor. No function could as yet be attributed to APRPs, although these neuropeptides occur abundantly in the corpora cardiaca, a neurohaemal organ from which many neuropeptides are released. Therefore, similar to GRF, a role as releasing factor may be envisaged for APRPs.

As neurons extend axons to their targets during development, growth cones must reorient their direction of migration in response to extracellular guidance cues. A variety of model systems have been employed in order to dissect the cellular and molecular mechanisms that underlie this complex process. One preparation, the developing grasshopper limb bud, has proved to offer a number of advantages in which to examine mechanisms of growth cone guidance and motility in vivo. First, the relatively large size of the embryonic nervous system allows for straightforward imaging of both fixed and live neurons in vivo. Second, the peripheral nerves generated in the limb bud are highly stereotyped. Third, intact embryos can be cultured for a period of days, allowing for fairly easy perturbations at precise developmental stages. Fourth, due to the ease of dissection, numerous cell biological and molecular techniques can be utilized in the limb bud. Finally, axon guidance molecules and mechanisms are conserved between grasshoppers and other organism, including vertebrates.

Hemolymph proteins in the locust, Locusta migratoria migratorioides infected with the fungus Metarhizium anisopliae var acridum were analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Under conditions that allowed locusts to thermoregulate, 2 proteins, ITB1 (ca. 18kDa) and ITB2 (ca. 13kDa) were induced 48h post inoculation. In contrast, under non-thermoregulating conditions, only 1 band, INTB1 (ca. 18kDa) was induced with similar molecular mass to ITB1. ITB1 and ITB2 were N-terminally sequenced but showed little homology to known proteins. The induction of hemolymphal proteins in infected, thermoregulating locusts and implication in insect immune defence are discussed.

Studies were conducted with two different doses of Metarhizium anisopliae var acridum to examine the effects on survival and reproductive potential of adult Schistocerca gregaria under conditions that either limited thermoregulation or enabled optimal thermoregulation. Adult S. gregaria infected with the fungal pathogen showed either a rapid and high mortality at relatively constant temperatures or a much reduced mortality and lengthened survival time when allowed to thermoregulate. Mortality rate varied from >90% after 10 days under constant temperature conditions to 66% after 70 days under optimal thermoregulatory conditions. Effects of infection on maturation and reproduction depended on the age of the adults at the time of inoculation, the nighttime temperature regime, the fungal dose, and the length of time of the monitoring period. No difference in reproductive behaviors in treated and control insects were found in one experiment that utilized older adults and was conducted over 25 days. In a second experiment with newly fledged locusts, differences in maturation rates and total reproductive output were observed due to infection. The results from these experiments are discussed in terms of the potential of M. anisopliae var acridum to alter the balance of insect endocrine systems and the importance of the assessment of behavioral changes and their impact on microbial control agents in the long term., (Copyright 2001 Academic Press.)

The brain-corpora cardiaca-corpora allata complex of insects is the physiological equivalent of the brain-hypophysis axis of vertebrates. In locusts there is only one corpus cardiacum as a result of fusion, while most other insect species have a pair of such glands. Like the pituitary of vertebrates, the corpus cardiacum consists of a glandular lobe and a neurohemal lobe. The glandular lobe synthesizes and releases adipokinetic hormones. In the neurohemal part many peptide hormones, which are produced in neurosecretory cells in the brain, are released into the hemolymph. The corpora allata, which have no counterpart in vertebrates, synthesize and release juvenile hormones. The control of the locust corpus cardiacum-corpora allata complex appears to be very complex. Numerous brain factors have been reported to have an effect on biosynthesis and release of juvenile hormone or adipokinetic hormone. Many neuropeptides are present in nerves projecting from the brain into the corpora cardiaca-corpora allata complex, the most important ones being neuroparsins, ovary maturating parsin, insulin-related peptide, diuretic peptide, tachykinins, FLRFamides, FXPRLamides, accessory gland myotropin I, crustacean cardioactive peptide, and schistostatins. In this paper, the cellular distribution, posttranslational processing, peptide-receptor interaction, and inactivation of these peptides are reviewed. In addition, the signal transduction pathways in the release of adipokinetic hormone and juvenile hormone from, respectively, the corpora cardiaca and corpora allata are discussed.

Endothelin-1-like immunoreactivity has been found in endocrine cells of the midgut of the desert locust Locusta migratoria. Several antisera have been directed against the whole molecule and its C-terminal sequence. Endothelin-1-immunoreactive cells are present in the main region of the midgut (ventriculus) and in the midgut caeca but not in the ampullae through which the malpighian tubules drain. Endothelin-1-like immunoreactivity colocalizes with FMRFa immunoreactivity in the cells of the main region of the midgut but not in those in the midgut caeca. Endothelin-1-immunoreactive cells are present not only in adults but also throughout the five instars of posthatching development.

Investigations into the behavioural and underlying physiological mechanisms of dietary selection are presented for the locust, Locusta migratoria. Locusts were fed for 4, 8 or 12 h on one of four chemically defined artificial diets: diet PC, which was nutritionally complete; diet P, containing no digestible carbohydrate; diet C, containing no protein; and diet O, which lacked both protein and digestible carbohydrate. Following this pretreatment, the locusts were provided with both the P and the C diet in a choice test. Detailed analyses of selection behaviour indicated that diets lacking a nutrient for which the insect was deficient were either rejected before a meal was initiated, or, if feeding commenced, eaten in meals of only short duration, while those containing the appropriate nutrients were accepted more readily and eaten in longer meals. Electrophysiological studies showed that this behaviour was paralleled by nutrient-specific changes in gustatory responsiveness. Locusts pretreated for 4h on C diet had increased gustatory responsiveness to stimulation with an amino acid mix, but not to sucrose, while insects fed on P diet showed increased responsiveness to stimulation with sucrose, but not to the amino acid mix. This result is consistent with earlier experiments in which levels of blood nutrients were shown to modulate taste responsiveness in the locust.

A neuropeptide which stimulates the motility of the cockroach hindgut has been isolated from an extract of 9000 brain-corpora cardiaca-corpora allata-subesophageal ganglion complexes of Locusta migratoria. Biological activity was monitored during HPLC purification by observing the myotropic effect of column fractions on the isolated hindgut of Leucophaea maderae. The primary structure of this myotropic peptide was established as a blocked 16-residue peptide: pGlu-Asp-Ser-Gly-Asp-Gly-Trp-Pro-Gln-Gln-Pro-Phe-Val-Pro-Arg-Leu-NH2. This novel locust peptide was designated as locustapyrokinin, or Lom-PK. Lom-PK was synthesized and shown to have chromatographic and biological properties identical to those of the native material. Lom-PK has a Phe-X-Pro-Arg-Leu-NH2 carboxy terminal in common with leucopyrokinin (or Lem-PK), a blocked myotropic neuropeptide isolated from the cockroach hindgut. The constituent amino acids of this C-terminal are important for biological activity on the Leucophaea hindgut. The primary structure of this novel insect peptide is, however, substantially different from Lem-PK at the amino-terminal sequence.

Proteinaceous factors from the corpora cardiaca (CC) and ventral ganglia (VG) stimulate KCl transport across voltage-clamped preparations of locust ilea in a dose-dependent manner. These factors may act via cAMP as the second messenger because this cyclic nucleotide and agents which increase its cellular level all mimic the stimulatory effects of CC and VG extracts. The CC and VG factors are apparently different compounds because they differ in the time course of ileal short-circuit current (Isc) initiated, in heat stability, and in extraction properties. Because saline extracts of the CC have similar effects on both ileal and rectal segments of locust hindgut, the ileal stimulant from the CC may be the chloride transport-stimulating hormone, described by Phillips et al. (Canad. J. Zool. 58, 1851-1860, 1980) which acts to stimulate rectal Isc.

The regulation of feeding in two insect groups, the locusts and the blowflies, is reviewed. The patterning of feeding is discussed first, and then the underlying mechanisms are considered in detail. The concept of "central excitability" is the key to understanding the controls of feeding. It represents a central nervous mechanism for tuning the sensitivity of the insect to food-related stimuli. The level of central excitation is influenced positively by food and also non-food stimuli, both from within and outside the animal, and negatively by deterrent stimuli and by feedbacks from peripheral systems such as stretch receptors on the gut wall, hormones and blood composition. Sustained gustatory input is required if feeding is to continue, once initiated. The level of central excitation present as a meal begins influences ingestion rate and the amounts of negative feedback tolerated before feeding stops. The duration of intervals between meals is then a function of a number of excitatory stimuli and of feeding-induced changes which reduce the likelihood of further feeding, e.g., hormone release, changes in nutrient composition or osmotic pressure of the blood, and gut distension. Other areas reviewed include changes in the regulation of feeding during the life history and the control of drinking.

The presence of binding sites for nonecdysteroid steroids was investigated in the cytosol of several tissues of the migratory locust, Locusta migratoria migratorioides. Binding of androgens was not observed. Most tissues, however, showed nonsaturable binding of estrogens and in some tissues saturable progestin binding could be demonstrated. A pregnenolone binder, that was found to be present in the male copulatory organ, was further studied. It showed a dissociation constant of 4.4 (+/- 1.6) X 10(-8) M. This is the first report of a nonecdysteroid steroid-binding factor in an insect tissue.

Nervous lobes of the migratory locust corpora cardiaca contain an antidiuretic (AD) factor which increases the fluid reabsorption of isolated recta in a dose-dependent manner. Several extraction procedures were developed and compared. The very satisfactory thermostability of the AD factor was demonstrated. By using a succession of different incubating media on the same preparation, we improved our bioassay. The developed variant is faster and more sensitive than the existing test and saves time and biological material.

Endogenous cyclic-AMP concentrations in Malpighian tubules of locusts increase as a result of diuretic hormone stimulation, and the level of increase is shown to be dependent on basal intracellular levels of cyclic-AMP. Furthermore, the increase in endogenous cyclic-AMP is in such excess as to cause excretion of high levels of cyclic-AMP in the urine.

Transection of the nervous connections (NCC I) between the brain and the corpus cardiacum (group 1) and injection of anti-brain antibodies inhibited vitellogenin synthesis and ovary development in virgin Locusta migratoria females. Removal of the neurosecretory material by these methods changed the titer and the electrophoretic pattern of hemolymph proteins, including vitellogenin. Transection of NCC I at some distance from the brain resulted in bulbs with 30-200 micron diameter which were filled with stainable neurosecretory material (group 2). Oocyte maturation was delayed in group 2 but normal eggs were laid. The same happened if group 1 animals received daily injections of JH-III. It is concluded that a humoral factor is involved in the activation of JH-III biosynthesis. The factor is transported through the NCC I and is released into the hemolymph as allatotropin.

A comparative ultrastructural study of insects was made whose primitive perisympathetic organs consisted either of a single median neurohemal formation per segment (Periplaneta americana) or of two transverse formations (Carausius morosus), or again, of three--one median and two transverse--as in Locusta migratoria. The results showed that the perisympathetic organs always comprised two types of structure (alpha and beta), even when a single formation exists. In Periplaneta and Carausius, both types were seen to coexist in the same neurohemal formation. In Locusta, however, they were separated, the alpha structure constituting the median organ and the beta structure, the two transverse organs. Each of the two structures have special features: the beta structure displays the usual characteristics of a loose neurohemal organ, i.e., it is penetrated by sinuses and has neurosecretory endings devoid of a glial coat; the alpha structure, on the contrary, forms dense compact organs whose endings are covered with a continuous glial layer. During the evolution, this last structure gradually separated from the other neurohemal formations. It was observed to correspond to a single type of neurosecretory cell, distinct from all other cell types by the particular mode of its release. These results for primitive perisympathetic organs confirm earlier findings for advanced organs. They indicate that both the alpha and beta structures are generally present in insects, thus showing their importance in the physiology of these animals.

The data derived from mating experiments demonstrate that mating has an accelerating effect on oviposition in the female Locusta migratoria. The effect of mating could be mimicked by injection of extracts of the male accessory reproductive gland. The oviposition-stimulating factor was localized in the opalescent gland of the male accessory gland and was transferred to the female via the spermatophore during copulation. Gel filtration of an extract of the opalescent gland revealed a 13,000 Da protein, which, when injected into virgin female locusts, could stimulate the oviposition rate to that seen in mated females. Extracts of the corpus cardiacum also stimulated oviposition when injected into virgin female locusts. This increase was not observably different from that seen after mating. The relevance of these findings will be discussed.

This study discusses the effects of a 1-hr period of flight on the peptidergic pars intercerebralis (PI)-corpus cardiacum storage part (CCS) system in male Locusta migratoria, particularly the effect on material in this system stained by a histochemical method for peptidergic neurosecretory material (NSM) or labeled by in vivo incorporation of radioactive amino acid molecules. By use of an automatic image analysis system a number of parameters of the stained or radioactively labeled substances were measured to quantify the flight-induced effects and to get information on the manner in which the neurosecretory cell bodies in the PI and their axonal endings in the CCS accommodate changing amounts of NSM. The CCS of flown locusts contained distinctly more stained and radioactively labeled substances than the CCS of unflown locusts. A tendency to similar differences was observed in the cluster of neurosecretory cell bodies in the PI. The results indicate that 1 hr flight inhibited the release of NSM by the PI-CCS system. After the onset of reduced release activity by flight, some NSM continued to be synthesized and transported from the PI to the CCS, gradually filling up and expanding the entire PI-CCS system, the NSM at the same time becoming more and more densely packed. It is concluded that the peptidergic PI-CCS system is not actively involved in the control of flight metabolism or flight behavior.