Effects of different stimulators of cGMP synthesis on lipid content in bovine oocytes matured in vitro

Bovine oocytes and blastocysts produced in vitro are frequently of lower quality and less cryotolerant than those produced in vivo, and greater accumulation of lipids in the cytoplasm has been pointed out as one of the reasons. In human adipocytes cGMP signaling through the activation of PKG appears to be involved in lipid metabolism, and components of this pathway have been detected in bovine cumulus-oocyte complexes (COCs). The aim of this study was to investigate the influence of this pathway on the lipid content in oocytes and expression of PLIN2 (a lipid metabolism-related gene) in cumulus cells. COCs were matured in vitro for 24 h with different stimulators of cGMP synthesis. The activation of soluble guanylyl cyclase (sGC) by Protoporphyrin IX reduced lipid content (22.7 FI) compared to control oocytes (36.45 FI; P <0.05). Stimulation of membrane guanylyl cyclase (mGC) with natriuretic peptides precursors A and C (NPPA and NPPC) had no effect (36.5 FI; P>0.05). When the PKG inhibitor KT5823 was associated with Protoporphyrin IX, its effect was reversed and lipid contents increased (52.71 FI; P<0.05). None of the stimulators of cGMP synthesis affected the expression of PLIN2 in cumulus cells. In conclusion, stimulation of sGC for cGMP synthesis promotes lipolytic activities in bovine oocytes matured in vitro and such effect is mediated by PKG. However, such effect may vary depending on the stimulus received and/or which synthesis enzyme was activated, as stimulation of mGC had no effects.     

Mechanistic insights on novel small molecule allosteric activators of cGMP-dependent protein kinase PKG1α

cGMP-dependent protein kinase (PKG) represents a compelling drug target for treatment of cardiovascular diseases. PKG1 is the major effector of beneficial cGMP signaling which is involved in smooth muscle relaxation and vascular tone, inhibition of platelet aggregation and signaling that leads to cardioprotection. In this study, a novel piperidine series of activators previously identified from an ultrahigh-throughput screen were validated to directly bind partially activated PKG1α and subsequently enhance its kinase activity in a concentration-dependent manner. Compounds from initial optimization efforts showed an ability to activate PKG1α independent of the endogenous activator, cGMP. We demonstrate these small molecule activators mimic the effect of cGMP on the kinetic parameters of PKG1α by positively modulating the K_M of the peptide substrate and negatively modulating the apparent K_M for ATP with increase in catalytic efficiency, k_cat. In addition, these compounds also allosterically modulate the binding affinity of cGMP for PKG1α by increasing the affinity of cGMP for the high-affinity binding site (CNB-A) and decreasing the affinity of cGMP for the low-affinity binding site (CNB-B). We show the mode of action of these activators involves binding to an allosteric site within the regulatory domain, near the CNB-B binding site. To the best of our knowledge, these are the first reported non-cGMP mimetic small molecules shown to directly activate PKG1α. Insights into the mechanism of action of these compounds will enable future development of cardioprotective compounds that function through novel modes of action for the treatment of cardiovascular diseases.     

Structural Basis of Cyclic Nucleotide Selectivity in cGMP-dependent Protein Kinase II

Membrane-bound cGMP-dependent protein kinase (PKG) II is a key regulator of bone growth, renin secretion, and memory formation. Despite its crucial physiological roles, little is known about its cyclic nucleotide selectivity mechanism due to a lack of structural information. Here, we find that the C-terminal cyclic nucleotide binding (CNB-B) domain of PKG II binds cGMP with higher affinity and selectivity when compared with its N-terminal CNB (CNB-A) domain. To understand the structural basis of cGMP selectivity, we solved co-crystal structures of the CNB domains with cyclic nucleotides. Our structures combined with mutagenesis demonstrate that the guanine-specific contacts at Asp-412 and Arg-415 of the αC-helix of CNB-B are crucial for cGMP selectivity and activation of PKG II. Structural comparison with the cGMP selective CNB domains of human PKG I and Plasmodium falciparum PKG (PfPKG) shows different contacts with the guanine moiety, revealing a unique cGMP selectivity mechanism for PKG II.     

Study of inheritance of feeding potential in natural populations of predatory coccinellid Cryptolaemus montrouzieri Mulsant using isofemale strains

The ability to feed on the prey is of great concern for the predatory insects, especially with regard to predatory coccinellid, Cryptolaemus montrouzieri Mulsant, which is mass reared and released into the field in large numbers to control the target pests. The variability associated with feeding potential is partly influenced by the genetic background of the insects and partly due to the environment, but the genetic basis of this trait is not yet fully understood in C. montrouzieri. The aim of this study was to identify the genetic basis of variation and heritability of this quantitative trait in natural populations of C. montrouzieri through isofemale heritability and parent–offspring regression. The regression analyses indicated that there was a significant linear relationship between progeny and their mothers for feeding potential.     

First study of the biology of Cryptolaemus montrouzieri and its potential to feed on the mealybug Dactylopius opuntiae (Hemiptera: Dactylopiidae) under laboratory conditions in Morocco

Abstract

Dactylopius opuntiae, is known as specific Opuntia cochineal in many countries around the world. This sap-sucking insect was first detected in Morocco in 2014. To address the problem, the feeding potential of different development stages of Cryptolaemus montrouzieri Mulsant, a biological control agent against mealybugs, was investigated on different development stages of D. opuntiae. Fourth instar larvae and adults of C. montrouzieri were the most voracious feeders on different instars of the mealybug. The numbers of mealybug eggs consumed by first, second, third and fourth instar larva and adults of C. montrouzieri were 36.18?±?1.84, 68.08?±?4.17, 280.55?±?5.41, 540.55?±?5.35, 6514.13?±?64.28, respectively. The numbers of mealybug nymphs consumed by the same stages of C. montrouzieri were 35.43?±?1.75, 67.73?±?3.88, 279.85?±?5.58, 539.63?±?5.08 and 6501.7?±?81.94 (first instars) and 34.83?±?1.20, 57.45?±?1.22, 83.80?±?1.92, 213.65?±?3.46 and 6013.23?±?35.46 (second instars), respectively. The corresponding figures for adult female mealybugs were 1.40?±?0.78, 10.65?±?1.83, 18.58?±?1.24, 25.23?±?1.10 and 197.15?±?3.29, respectively. The egg, larval, prepupal, pupal and adult stages occupied 3.36–3.69, 20.21–27.59, 1.31–1.59, 10.62–10.72 and 96.10–102.51?days, respectively when the coccinellid was reared on different stages of D. opuntiae. The results indicate that C. montrouzieri has the potential to be used as a biocontrol agent in Morocco.     

Effects of three mealybug species on the development,survivorship and reproduction of the predatory lady beetle Cryptolaemus montrouzieri Mulsant

The lady beetle Cryptolaemus montrouzieri Mulsant (Coleoptera: Coccinellidae) is an important predator of mealybugs. The development, survivorship, longevity and reproduction of C. montrouzieri feeding on three different mealybug species [Dysmicoccus neobrevipes Beardsley, Ferrisia virgata Cockerell and Planococcus minor (Maskell)] were investigated in the laboratory at 26 ± 1°C, 75-–90% RH and 14:10 (L:D) h photoperiod. Results indicated that, when feeding on different mealybugs, no significant differences were observed between developmental periods and survivorship of C. montrouzieri (from egg to adult), but differences were recorded between the sex ratios, preovipositional periods, adult longevities and reproduction of the differently treated lady beetle populations. The highest sex ratio (0.56), the longest preovipositional period (6.6 days) and adult longevity (84.8 days for females and 93.9 days for males), and the maximum fecundity (659.0 eggs/female) of C. montrouzieri were recorded when feeding on F. virgata. Moreover, C. montrouzieri had a high net reproductive rate (313.66), intrinsic rate of increase (0.0816) and finite rate of increase (1.085) when feeding on F. virgata. Results indicated that the population growth of C. montrouzieri may increase faster when feeding on F. virgata than feeding on either of the other two mealybugs.     

cGMP/Protein Kinase G Signaling Suppresses Inositol 1,4,5-Trisphosphate Receptor Phosphorylation and Promotes Endoplasmic Reticulum Stress in Photoreceptors of Cyclic Nucleotide-gated Channel-deficient Mice

Photoreceptor cyclic nucleotide-gated (CNG) channels play a pivotal role in phototransduction. Mutations in the cone CNG channel subunits CNGA3 and CNGB3 are associated with achromatopsia and cone dystrophies. We have shown endoplasmic reticulum (ER) stress-associated apoptotic cone death and increased phosphorylation of the ER Ca²⁺ channel inositol 1,4,5-trisphosphate receptor 1 (IP₃R1) in CNG channel-deficient mice. We also presented a remarkable elevation of cGMP and an increased activity of the cGMP-dependent protein kinase (protein kinase G, PKG) in CNG channel deficiency. This work investigated whether cGMP/PKG signaling regulates ER stress and IP₃R1 phosphorylation in CNG channel-deficient cones. Treatment with PKG inhibitor and deletion of guanylate cyclase-1 (GC1), the enzyme producing cGMP in cones, were used to suppress cGMP/PKG signaling in cone-dominant Cnga3^−/−/Nrl^−/− mice. We found that treatment with PKG inhibitor or deletion of GC1 effectively reduced apoptotic cone death, increased expression levels of cone proteins, and decreased activation of Müller glial cells. Furthermore, we observed significantly increased phosphorylation of IP₃R1 and reduced ER stress. Our findings demonstrate a role of cGMP/PKG signaling in ER stress and ER Ca²⁺ channel regulation and provide insights into the mechanism of cone degeneration in CNG channel deficiency.     

The locust foraging gene

Our knowledge of how genes act on the nervous system in response to the environment to generate behavioral plasticity is limited. A number of recent advancements in this area concern food‐related behaviors and a specific gene family called foraging (for), which encodes a cGMP‐dependent protein kinase (PKG). The desert locust (Schistocerca gregaria) is notorious for its destructive feeding and long‐term migratory behavior. Locust phase polyphenism is an extreme example of environmentally induced behavioral plasticity. In response to changes in population density, locusts dramatically alter their behavior, from solitary and relatively sedentary behavior to active aggregation and swarming. Very little is known about the molecular and genetic basis of this striking behavioral phenomenon. Here we initiated studies into the locust for gene by identifying, cloning, and studying expression of the gene in the locust brain. We determined the phylogenetic relationships between the locust PKG and other known PKG proteins in insects. FOR expression was found to be confined to neurons of the anterior midline of the brain, the pars intercerebralis. Our results suggest that differences in PKG enzyme activity are correlated to well‐established phase‐related behavioral differences. These results lay the groundwork for functional studies of the locust for gene and its possible relations to locust phase polyphenism. © 2010 Wiley Periodicals, Inc.     

Cofilin phosphorylation is involved in nitric oxide/cGMP-mediated nociception

There is convincing evidence that nitric oxide (NO), cGMP and cGMP-dependent protein kinase I (PKG-I) are involved in the development of hyperalgesia in response to noxious stimuli. However, downstream target proteins contributing to nociception have not been completely identified so far. Several reports indicate a role of the NO/cGMP/PKG cascade in the regulation of neurite outgrowth which is suggested to be involved in specific mechanisms of nociception. Since neurite outgrowth is strongly dependent on modulation of cytoskeleton proteins we were interested in the impact of PKG-I activation on the actin cytoskeleton and its role in inflammatory hyperalgesia. Therefore we investigated the actin-destabilising protein cofilin and its NO-dependent effects in vitro in primary neuronal cultures as well as in vivo in the zymosan-induced paw inflammation model in rats. In primary neurons from rats, treatment with the PKG-I activator 8-Br-cGMP induced a time-dependent phosphorylation of cofilin and significantly increased neurite outgrowth. Further functional analysis revealed that the underlying signal transduction pathways involve activation of the Rho-GTPases RhoA, Rac1 and Cdc42 and their corresponding downstream targets Rho-kinase (ROCK) and p21-activated kinase (PAK). In vivo, treatment of rats with the NO-synthase inhibitor l-NAME and the ROCK-inhibitor Y-27632, respectively, led to a significant decrease of cofilin phosphorylation in the spinal cord and resulted in antinociceptive effects in a model of inflammatory hyperalgesia. Our results suggest that cofilin represents a downstream target of NO/cGMP/PKG signal transduction in neurons thus indicating that it is involved in NO-mediated nociception.     

Type 2 cGMP-dependent protein kinase regulates homeostasis by blocking c-Jun N-terminal kinase in the colon epithelium

Analysis of knockout animals indicates that 3′,5′cyclic guanosine monophosphate (cGMP) has an important role in gut homeostasis but the signaling mechanism is not known. The goals of this study were to test whether increasing cGMP could affect colon homeostasis and determine the mechanism. We increased cGMP in the gut of Prkg2^+/+ and Prkg2^−/− mice by treating with the PDE5 inhibitor Vardenafil (IP). Proliferation, differentiation and apoptosis in the colon mucosa were then quantitated. Vardenafil (Vard) treatment increased cGMP in colon mucosa of all mice, but reduced proliferation and apoptosis, and increased differentiation only in Prkg2^+/+ mice. Vard and cGMP treatment also increased dual specificity protein phosphatase 10 (DUSP10) expression and reduced phospho-c-Jun N-terminal kinase (JNK) levels in the colon mucosa of Prkg2^+/+ but not Prkg2^−/− mice. Treatment of Prkg2^−/− mice with the JNK inhibitor SP600125 reversed the defective homeostasis observed in these animals. Activation of protein kinase G2 (PKG2) in goblet-like LS174T cells increased DUSP10 expression and reduced JNK activity. PKG2 also increased goblet cell-specific MUC2 expression in LS174T cells, and this process was blocked by DUSP10-specific siRNA. The ability of cGMP signaling to inhibit JNK-induced apoptosis in vivo was demonstrated using dextran sodium sulfate (DSS) to stress the colon epithelium. Vard was a potent inhibitor of DSS-induced epithelial apoptosis, and significantly blocked pathological endpoints in this model of experimental colitis. In conclusion, Vard treatment activates cGMP signaling in the colon epithelium. Increased PKG2 activity alters homeostasis by suppressing proliferation and apoptosis while promoting differentiation. The PKG2-dependent mechanism was shown to involve increased DUSP10 and subsequent inhibition of JNK activity.     

Physiological effects of compensatory growth during the larval stage of the ladybird,Cryptolaemus montrouzieri

The growth rate of insects may vary in response to shifty environments. They may achieve compensatory growth after a period of food restriction followed by ad libitum food, which may further affect the reproductive performance and lifespan of the resulting phenotypes. However, little is known about the physiological mechanisms associated with such growth acceleration in insects. The present study examined the metabolic rate, the antioxidant enzyme activity and the gene expression of adult Cryptolaemus montrouzieri (Coleoptera: Coccinellidae) after experiencing compensatory growth during its larval stages. Starved C. montrouzieri individuals achieved a similar developmental time and adult body mass as those supplied with ad libitum food during their entire larval stage, indicating that compensatory growth occurred as a result of the switch in larval food regime. Further, the compensatory growth was found to exert effects on the physiological functions of C. montrouzieri, in terms of its metabolic rates and enzyme activities. The adults undergoing compensatory growth were characterized by a higher metabolic rate, a lower activity of the antioxidant enzymes glutathione reductase, catalase, and superoxide dismutase and a lower gene expression of P450 and trehalase. Taken together, the results indicate that although compensatory growth following food restriction in early larval life prevents developmental delay and body mass loss, the resulting adults may encounter physiological challenges affecting their fitness.     

Mechanism of cAMP Partial Agonism in Protein Kinase G (PKG)

Protein kinase G (PKG) is a major receptor of cGMP and controls signaling pathways often distinct from those regulated by cAMP. Hence, the selective activation of PKG by cGMP versus cAMP is critical. However, the mechanism of cGMP-versus-cAMP selectivity is only limitedly understood. Although the C-terminal cyclic nucleotide-binding domain B of PKG binds cGMP with higher affinity than cAMP, the intracellular concentrations of cAMP are typically higher than those of cGMP, suggesting that the cGMP-versus-cAMP selectivity of PKG is not controlled uniquely through affinities. Here, we show that cAMP is a partial agonist for PKG, and we elucidate the mechanism for cAMP partial agonism through the comparative NMR analysis of the apo, cGMP-, and cAMP-bound forms of the PKG cyclic nucleotide-binding domain B. We show that although cGMP activation is adequately explained by a two-state conformational selection model, the partial agonism of cAMP arises from the sampling of a third, partially autoinhibited state.     

Co-crystal structures of PKG Iβ (92-227) with cGMP and cAMP reveal the molecular details of cyclic-nucleotide binding

Background

Cyclic GMP-dependent protein kinases (PKGs) are central mediators of the NO-cGMP signaling pathway and phosphorylate downstream substrates that are crucial for regulating smooth muscle tone, platelet activation, nociception and memory formation. As one of the main receptors for cGMP, PKGs mediate most of the effects of cGMP elevating drugs, such as nitric oxide-releasing agents and phosphodiesterase inhibitors which are used for the treatment of angina pectoris and erectile dysfunction, respectively.

Methodology/Principal Findings

We have investigated the mechanism of cyclic nucleotide binding to PKG by determining crystal structures of the amino-terminal cyclic nucleotide-binding domain (CNBD-A) of human PKG I bound to either cGMP or cAMP. We also determined the structure of CNBD-A in the absence of bound nucleotide. The crystal structures of CNBD-A with bound cAMP or cGMP reveal that cAMP binds in either syn or anti configurations whereas cGMP binds only in a syn configuration, with a conserved threonine residue anchoring both cyclic phosphate and guanine moieties. The structure of CNBD-A in the absence of bound cyclic nucleotide was similar to that of the cyclic nucleotide bound structures. Surprisingly, isothermal titration calorimetry experiments demonstrated that CNBD-A binds both cGMP and cAMP with a relatively high affinity, showing an approximately two-fold preference for cGMP.

Conclusions/Significance

Our findings suggest that CNBD-A binds cGMP in the syn conformation through its interaction with Thr193 and an unusual cis-peptide forming residues Leu172 and Cys173. Although these studies provide the first structural insights into cyclic nucleotide binding to PKG, our ITC results show only a two-fold preference for cGMP, indicating that other domains are required for the previously reported cyclic nucleotide selectivity.     

Invasion of hepatocytes by Plasmodium sporozoites requires cGMP‐dependent protein kinase and calcium dependent protein kinase 4

Invasion of hepatocytes by sporozoites is essential for Plasmodium to initiate infection of the mammalian host. The parasite's subsequent intracellular differentiation in the liver is the first developmental step of its mammalian cycle. Despite their biological significance, surprisingly little is known of the signalling pathways required for sporozoite invasion. We report that sporozoite invasion of hepatocytes requires signalling through two second‐messengers – cGMP mediated by the parasite's cGMP‐dependent protein kinase (PKG), and Ca²⁺, mediated by the parasite's calcium‐dependent protein kinase 4 (CDPK4). Sporozoites expressing a mutated form of Plasmodium berghei PKG or carrying a deletion of the CDPK4 gene are defective in invasion of hepatocytes. Using specific and potent inhibitors of Plasmodium PKG and CDPK4, we demonstrate that PKG and CDPK4 are required for sporozoite motility, and that PKG regulates the secretion of TRAP, an adhesin that is essential for motility. Chemical inhibition of PKG decreases parasite egress from hepatocytes by inhibiting either the formation or release of merosomes. In contrast, genetic inhibition of CDPK4 does not significantly decrease the number of merosomes. By revealing the requirement for PKG and CDPK4 in Plasmodium sporozoite invasion, our work enables a better understanding of kinase pathways that act in different Plasmodium stages.     

Up‐regulation of Tyrosinase Gene by Nitric Oxide in Human Melanocytes

Ultraviolet light (UV) radiation causes skin‐tanning, which is thought to be mediated by stimulating the release of melanogenic factors from keratinocytes as well as other cells. Nitric oxide (NO) has been reported to be generated after UV radiation and to stimulate melanocytes as one of the melanogens. In a previous experiment by another group on melanogenesis induced by NO, increases in both tyrosinase activity and tyrosinase protein levels were observed after daily stimulation of NO for 4 days. In the present study, we investigated tyrosinase gene expression within the first 24 hr of NO‐induced melanogenesis. Tyrosinase mRNA expression was found to be induced 2 hr after a single treatment with S‐nitroso‐N‐acetyl‐ l ‐arginine. An increase of tyrosinase activity was also detected time‐dependently within the 24‐hr period, accompanied by an increase of tyrosinase protein levels. The induction of mRNA expression was suppressed by a cyclic guanosine 3′,5′‐monophosphate (cGMP)‐dependent protein kinase (cGMP/PKG) inhibitor. These results suggest that the enhancement of tyrosinase gene expression via the cGMP pathway may be a primary mechanism for NO‐induced melanogenesis.     

Essential role of nitric oxide in acute ischemic preconditioning: S-Nitros(yl)ation versus sGC/cGMP/PKG signaling?

Nitric oxide (NO) plays an important role in acute ischemic preconditioning (IPC). In addition to activating soluble guanylyl cyclase (sGC)/cyclic guanosine monophosphate (cGMP)/protein kinase G (PKG) signaling pathways, NO-mediated protein S-nitros(yl)ation (SNO) has been recently shown to play an essential role in cardioprotection against ischemia–reperfusion (I/R) injury. In our previous studies, we have shown that IPC-induced cardioprotection could be blocked by treatment with either N-nitro-L-arginine methyl ester (L-NAME, a constitutive NO synthase inhibitor) or ascorbate (a reducing agent to decompose SNO). To clarify NO-mediated sGC/cGMP/PKG-dependent or -independent (i.e., SNO) signaling involved in IPC-induced cardioprotection, mouse hearts were Langendorff-perfused in the dark to prevent SNO decomposition by light exposure. Treatment with 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ, a highly selective inhibitor of sGC) or KT5823 (a potent and selective inhibitor of PKG) did not abolish IPC-induced acute protection, suggesting that the sGC/cGMP/PKG signaling pathway does not play an important role in NO-mediated cardioprotective signaling during acute IPC. In addition, treatment with ODQ in IPC hearts provided an additional protective effect on functional recovery, in parallel with a higher SNO level in these ODQ+IPC hearts. In conclusion, these results suggest that the protective effect of NO is not related primarily to activation of the sGC/cGMP/PKG signaling pathway, but rather through SNO signaling in IPC-induced acute cardioprotection.     

Cyclic nucleotide selectivity of protein kinase G isozymes

The intrinsic activity of the C‐terminal catalytic (C) domain of cyclic guanosine monophosphate (cGMP)‐dependent protein kinases (PKG) is inhibited by interactions with the N‐terminal regulatory (R) domain. Selective binding of cGMP to cyclic nucleotide binding (CNB) domains within the R‐domain disrupts the inhibitory R–C interaction, leading to the release and activation of the C‐domain. Affinity measurements of mammalian and plasmodium PKG CNB domains reveal different degrees of cyclic nucleotide affinity and selectivity; the CNB domains adjacent to the C‐domain are more cGMP selective and therefore critical for cGMP‐dependent activation. Crystal structures of isolated CNB domains in the presence and absence of cyclic nucleotides reveal isozyme‐specific contacts that explain cyclic nucleotide selectivity and conformational changes that accompany CNB. Crystal structures of tandem CNB domains identify two types of CNB‐mediated dimeric contacts that indicate cGMP‐driven reorganization of domain–domain interfaces that include large conformational changes. Here, we review the available structural and functional information of PKG CNB domains that further advance our understanding of cGMP mediated regulation and activation of PKG isozymes.     

<Emphasis Type="Italic">egl</Emphasis>-<Emphasis Type="Italic">4</Emphasis> modulates electroconvulsive seizure duration in <Emphasis Type="Italic">C. elegans</Emphasis>

Increased neuronal excitability causes seizures with debilitating symptoms. Effective and noninvasive treatments are limited for easing symptoms, partially due to the complexity of the disorder and lack of knowledge of specific molecular faults. An unexplored, novel target for seizure therapeutics is the cGMP/protein kinase G (PKG) pathway, which targets downstream K⁺ channels, a mechanism similar to Retigabine, a recently FDA-approved antiepileptic drug. Our results demonstrate that increased PKG activity decreased seizure duration in C. elegans utilizing a recently developed electroconvulsive seizure assay. While the fly is a well-established seizure model, C. elegans are an ideal yet unexploited model which easily uptakes drugs and can be utilized for high-throughput screens. In this study, we show that treating the worms with either a potassium channel opener, Retigabine or published pharmaceuticals that increase PKG activity, significantly reduces seizure recovery times. Our results suggest that PKG signaling modulates downstream K⁺ channel conductance to control seizure recovery time in C. elegans. Hence, we provide powerful evidence, suggesting that pharmacological manipulation of the PKG signaling cascade may control seizure duration across phyla.     

Dispersal potential of native and exotic predatory ladybirds as measured by a computer-monitored flight mill

The performance of three species of predatory ladybirds was compared in a flight mill and the effect of diet on their flight parameters was tested. The invasive ladybird Harmonia axyridis Pallas (Coleoptera: Coccinellidae) outperformed Cryptolaemus montrouzieri Mulsant (Coleoptera: Coccinellidae) and Adalia bipunctata (L.) (Coleoptera: Coccinellidae) in terms of flight distance, duration and velocity. Harmonia axyridis flew at least two times further, needed three times less breaks and flew two times faster than C. montrouzieri and A. bipunctata fed the same diet. Ladybirds reared on eggs of Ephestia kuehniella (Zeller) (Lepidoptera: Pyralidae) performed better than their counterparts reared on natural prey (aphids for H. axyridis and A. bipunctata, mealybugs for C. montrouzieri). The findings of this study indicate that comparative flight studies can be useful to identify candidate biocontrol agents with pronounced dispersal abilities and thus can yield significant evidence to be used in an environmental risk assessment. However, it also demonstrates that variability related to mass rearing conditions should not be ignored when standardizing a risk assessment procedure for candidate biocontrol agents.     

Establishment and yearly/seasonal occurrence of the exotic coccidophagous ladybird <Emphasis Type="Italic">Cryptolaemus montrouzieri</Emphasis> (Coleoptera: Coccinellidae) in citrus groves in Shizuoka City,central Japan: a 5-year survey on adult numbers

Cryptolaemus montrouzieri Mulsant (Coleoptera: Coccinellidae) is a ladybird native to Australia, preying on mealybugs and soft scales, and has been utilized worldwide as a biological control agent. It has long been recognized that C. montrouzieri that was introduced into the main island of Japan had failed to become established. The present study monitored yearly and seasonal occurrence of C. montrouzieri adults in citrus groves at Shizuoka Prefectural Fruit Tree Research Center in Shizuoka City, central Japan in 2008–2012 by using sticky traps and beating citrus trees. Adults of C. montrouzieri were continuously captured for 5 and 4 years in a pesticide-free citrus grove and a neighboring reduced-pesticide grove, respectively. Larvae of C. montrouzieri were observed consuming a cottony scale, Pulvinaria aurantii Cockerell, on citrus trees. These results provide unequivocal evidence for the ladybird’s establishment in central Japan. The number of trapped ladybird adults exhibited four peaks a year: in mid-April, early to late June, mid-August, and late September to early October. Adult numbers in each grove varied largely across years, showing a great increase followed by a rapid decline during a period of 4 years. Factors affecting the seasonal/yearly occurrence of C. montrouzieri adults in citrus groves are discussed.