Development of a Method for Rearing Nezara viridula (Heteroptera: Pentatomidae) on a Semi-solid Artificial Diet

A method for rearing the southern green stinkbug, (Nezara viridula L.) (Heteroptera: Pentatomidae), using a modified lygus semi-solid artificial diet was developed. First to second-instar nymph were reared in a density of 631.5 ± 125.05 eggs per Petri-dish (4 cm deep × 15 cm diam). Second instar to adult were reared in a density of 535.0 ± 112.46 s instar nymphs per rearing cage (43 × 28 × 9 cm). Mating and oviposition occurred in popup rearing cages (30 × 30 cm), each holding 60–90 mixed sex adults of similar age. Adults emerged 35.88 ± 2.13 d after oviposition and survived for an average of 43.09 ± 9.53 d. On average, adults laid 223.95 ± 69.88 eggs in their lifetime, for a total production of 8,099 ± 1,277 fertile eggs/oviposition cage. Egg fertility was 77.93% ± 16.28. Egg masses held in petri-dishes had a total hatchability of 79.38% ± 20.03. Mortality of early nymphs in petri-dishes was 0.64% ± 0.12 for the first instar and 1.37% ± 0.45 for second instar. Late nymphal mortality in rearing cages was 1.41% ± 0.10, 3.47% ± 1.27, and 4.72% ± 1.29 for the third, fourth, and fifth instars, respectively. Survivorship from nymphs to adults was 88.48% ± 2.76. Using artificial diet for rearing N. viridula could reduce cost by avoiding time-consuming issues with daily feeding fresh natural hosts and insect manipulation. It could increase reliability and simplicity of bug production, which should facilitate mass rearing of its biological control agents.     

Economic injury levels for southern green stink bugs (Hemiptera: Pentatomidae) in R7 growth stage soybeans

Stink bugs, primarily southern green stink bug, Nezara viridula (Hemiptera: Pentatomidae), are a major pest complex of soybeans (Glycine max) throughout the southern United States. Densities sometimes peak during late R6 and R7 soybean growth stages when soybeans are approaching physiology maturity and the rate of injury from stink bugs is reduced. Field cage trials were conducted from 2005 to 2008 to examine the type and extent of soybean damage caused by southern green stink bugs during the R7 growth stage. The yield response was variable, but overall was not significant. The impact of southern green stink bugs on quality was more consistent. Test weight decreased, and heat damage and total damage increased as stink bug density increased. Based on these data, three economic injury models were developed using different assumptions. The model that assumes no yield loss, does not predict economic injury within the range of stink bug densities tested. However, if the statistically non-significant yield losses are accepted as real, then the models suggest that the southern green stink bug economic injury level and action threshold for soybeans during R7 stage is generally between nine and 15 stink bugs per row m.     

Adaptation of Habrobracon hebetor (Hymenoptera: Braconidae) to Rearing on Ephestia kuehniella (Lepidoptera: Pyralidae) and Helicoverpa armigera (Lepidoptera: Noctuidae)

Food characteristics strongly regulate digestive enzymatic activity of insects through direct influences on their midgut mechanisms. Insect performance is better on diets that contain nutrients in proportions that fit its digestive enzymes. Little is known about the influences of rearing history on parasitism success of Habrobracon hebetor Say. This research focused on the effect of nutrient regulation on survival, development, and parasitism of H. hebetor. Life history and digestive enzyme activity of fourth-stage larvae of H. hebetor were studied when reared on Ephestia kuehniella Zeller. This parasitoid was then introduced to Helicoverpa armigera (Hübner), and above-mentioned parameters were also studied in the first and fourth generations after transfer. In term of parasitism success, H. hebetor preferred E. kuehniella over He. armigera. When the first and fourth generations of He. armigera-reared H. hebetor were compared, the rearing history affected the life history and enzymatic activity of the parasitoid. A better performance of H. hebetor was achieved after it was reared on He. armigera for the four generations. Because, digestive α-amylase and general protease of the parasitoid were matched with the new host, it used reserve energy for a better performance. Thus, a better performance of H. hebetor could be obtained when the parasitoid was reared on its original host for at least four generations.     

Spatiotemporal Distribution of Two Euschistus spp. Stink Bugs (Hemiptera: Pentatomidae) in Southeastern Farmscapes

Stink bugs (Hemiptera: Pentatomidae) are ubiquitous, cryptic, phytophagous pests that are found in many crops. In agroecosystems, individuals disperse from adjacent noncrop hosts and tend to aggregate or cluster within fields. In this study, we characterized the distribution of Euschistus servus (Say) and Euschistus tristigmus (Say) (Hemiptera: Pentatomidae) over 2 yr at three southeastern United States farmscapes. Stink bugs were captured in pheromone-baited traps, and Spatial Analysis by Distance Indices (SADIE) used to identify the location of significant aggregations by habitat type and season. Euschistus servus adults were more likely to be captured in pecan orchards, cotton, other crops, or unmanaged habitats than in woodland habitats. Significant aggregations of E. servus were detected in a variety of habitats including pecan, corn, cotton, peanut, and tobacco, as well as fallow and hay fields, pastures, and hedgerows. Fewer adult E. tristigmus were captured than E. servus adults, and E. tristigmus adults were typically trapped and aggregated in woodland habitats. The resulting data provide an important understanding regarding the seasonal movement and relative abundance levels of stink bug populations, which are critical to the development of integrated pest management strategies.     

Intraspecific Variation of Eysarcoris guttigerus (Hemiptera: Pentatomidae) in Japanese Southwest Population Based on Mitochondrial DNA

The white-spotted globular bug Eysarcoris guttigerus (Thunberg) (Hemiptera: Pentatomidae) is widely distributed in East Asia and the Pacific region. In Japan, the species is found in grassy or composite weeds in the western area of the main islands and Ryukyu Islands of Japan. One notable characteristic of the Eysarcoris genus is the two white spots on the scutellum. This is not the case with the Ishigaki Island population, however, which sports red spots instead of white, suggesting that intraspecific variation exists in the species. Therefore, we investigated intraspecific variation in E. guttigerus using mitochondrial NADH dehydrogenase subunit 2 (ND2), cytochrome oxidase subunit 1 (CO1), cytochrome b (Cytb), tRNA-Serine (tRNA^ser), NADH dehydrogenase subunit 1 (ND1), and 16S ribosomal RNA (16SrRNA) genes from 13 populations of Japan. The obtained maximum likelihood phylogenetic tree was divided into three groups—Group 1: Mainland, Group 2: Central Ryukyu Islands (Okinawa-Amamioshima Islands), and Group 3: South Ryukyu Islands (Ishigaki Island). The Ishigaki population was significantly separated from the other populations with consistent differences in spot color. The estimated period of divergence between the Ishigaki population and the other populations was consistent with the period of formation of the Kerama Gap in the Ryukyu arc. Thus, the process of formation of the Kerama Gap may have influenced the intraspecific variation of E. guttigerus.     

Efficacy of Plectranthus glandulosus (Lamiaceae) and Callistemon rigidus (Myrtaceae) Leaf Extract Fractions to Callosobruchus maculatus (Coleoptera: Bruchidae)

As part of on-going efforts to use eco-friendly alternatives to chemical pesticides, methanol crude extracts of Plectranthus glandulosus and Callistemon rigidus leaves were sequentially fractionated in hexane, chloroform, ethyl acetate, and methanol to establish the most active fraction(s) against Callosobruchus maculatus in cowpea. Cowpea seeds (25 g) were treated with 0.5, 1, 2, and 4 g/kg of extract to evaluate the contact toxicity and F₁ progeny production of the beetles in the laboratory. Mortality was recorded 1, 3, and 7 d postexposure. P. glandulosus hexane fraction was more toxic than the other fractions recording 100% mortality at 4 g/kg, within 7 d with LC₅₀ of 0.39 g/kg. Hexane fraction of C. rigidus showed superior toxicity, causing 100% mortality at 4 g/kg within only 1 d of exposure with LC₅₀ of 1.02 g/kg. All the fractions greatly reduced progeny emergence, with C. rigidus hexane fraction being the best progeny inhibitor. Fractions of P. glandulosus and C. rigidus leaves had sufficient efficacy to be a component of storage pest management package for C. maculatus.     

Abundance and Frequency of the Asiatic Oak Weevil (Coleoptera: Curculionidae) and Defoliation on American,Chinese, and Hybrid Chestnut (Castanea)

The Asiatic oak weevil, Cyrtepistomus castaneus Roelofs (Coleoptera: Curculionidae), is a nonnative defoliator of trees in the Fagaceae family in the United States but has not been studied on Castanea species in the southern Appalachian Mountains. Planted trees of Castanea dentata (Marsh.) Borkh. (Fagales: Fagaceae), Castanea mollissima Blume (Fagales: Fagaceae), and four hybrid breeding generations were evaluated in 2012 for insect defoliation and C. castaneus abundance and frequency. Defoliation was visually assessed throughout the growing season at two sites in the southern Appalachian Mountains (western North Carolina and eastern Tennessee). C. castaneus abundance and frequency were monitored on trees using beat sheets and emergence was recorded from ground traps. Asiatic oak weevils were more abundant and more frequently collected on American chestnut (Ca. dentata) and its most closely related BC₃F₃ hybrid generation than on the Asian species Ca. mollissima. In most months, C. castaneus colonization of hybrid generations was not significantly different than colonization of parental species. Frequency data for C. castaneus suggested that adults were distributed relatively evenly throughout the study sites rather than in dense clusters. Emergence of C. castaneus was significantly higher under a canopy dominated by Quercus species than under non-Quercus species or open sky. C. castaneus emergence began in May and peaked in late June and early July. These results may be useful for resource managers trying to restore blight-resistant chestnut to the Southern Appalachians while minimizing herbivory by insect pests.     

Terpenoids from the octocorals Menella sp. (Plexauridae) and Lobophytum crassum (Alcyonacea)

A new germacrane-type sesquiterpenoid, menelloide E (1), and a new cembrane-type diterpenoid, lobocrassin F (2), were isolated from the octocorals Menella sp. and Lobophytum crassum, respectively. The structures of terpenoids 1 and 2 were determined by spectroscopic and chemical methods and compound 2 was found to display a significant inhibitory effect on the release of elastase by human neutrophils.     

Influence of Constant Temperature on Reproductive Parameters of Holotrichia oblita (Coleoptera: Scarabaeidae)

Holotrichia oblita Faldermann (Coleoptera: Scarabaeidae) is a major pest both in field crops and forests because the larvae could eat the roots of most crops in the field, and the adults damage the leaves of trees and field crops. In this study, we focused on the effects of temperature on H. oblita reproductive parameters. The results indicated H. oblita female adults at 25°C could lay more eggs (84.0 eggs per female) and have the shortest preoviposition period (19.1 d), the greatest oviposition rate (2.8 eggs per female per 3 d), and largest percentage of life span spent in oviposition (59.5%). The longevity and the time to 50% egg laying decreased with increasing temperature, and female longevity was always longer than male longevity. The preoviposition and postoviposition period decreased with increasing temperature from 15 to 25°C and then increased when the temperature increased from 25 to 30°C. These results show that 25°C is the optimal temperature for reproduction of H. oblita.     

Creams Formulated with Ocimum gratissimum L. and Lantana camara L. Crude Extracts and Fractions as Mosquito Repellents Against Aedes aegypti L. (Diptera: Culicidae)

Mosquitoes are the most deadly vectors of parasites that cause diseases such as malaria, yellow fever, and filariasis. In view of the recent increased interest in developing plant origin insecticides as an alternative to chemical insecticides, the objective of this study was to determine the repellent activity of creams formulated with methanol crude extract (MCE), hexane fraction (HF), and ethyl acetate fractions (EAFs) of Ocimum gratissimum and Lantana camara leaves in single and combined actions against female Aedes aegypti. Evaluation was carried out in the net cages (30 by 30 by 30 cm) containing 60 blood-starved female mosquitoes each and were assayed in the laboratory condition following World Health Organization 2009 protocol. All formulations (single and mixture) were applied at 2, 4, 6, and 8 mg/cm² in the exposed area of human hands. Only acetone + white soft paraffin served as negative control and odomos (12% DEET) as positive control. All the formulations presented good protection against mosquito bites without any allergic reaction by the human volunteers. The repellent activity was dependent on the strength of the extracts and fractions. Among the tested formulations, the maximum protection time was observed in MCE (120 min) and EAF (150 min) of O. gratissimum; MCE:MCE (150 min) and HF:HF (120 min) mixtures of both plants. In addition, MCE:MCE and HF:HF mixtures from both plants showed possible synergistic effect. From the results, the combination of O. gratissimum and L. camara to formulate natural mosquito repellent using small amount of extracts can be encouraging to be an alternative to conventional DEET.     

Identification and Expression of Two Novel Cytochrome P450 Genes,CYP6CV1 and CYP9A38, in Cnaphalocrocis medinalis (Lepidoptera: Pyralidae)

Cnaphalocrocis medinalis Güenée can cause severe losses in rice. Cytochrome P450s play crucial roles in the metabolism of allelochemicals in herbivorous insects. Two novel P450 cDNAs, CYP6CV1 and CYP9A38, were cloned from the midgut of C. medinalis. CYP6CV1 encodes a protein of 500 amino acid residues, while CYP9A38-predicted protein has 531 amino acid residues. Both cDNA-predicted proteins contain the conserved functional domains for all P450s. Phylogenetic analyses showed that CYP6CV1 is grouped in the cluster containing CYP6B members, while CYP9A38 is in the cluster including CYP9 members. However, both clusters are contained in the same higher lineage. Homologous analysis revealed that CYP6CV1 is most similar to CYP6B8, CYP6B7, CYP6B6, CYP6B2, and CYP6B4 with the highest amino acid identity of 41%. CYP9A38 is closest to CYP9A17, CYP9A21, CYP9A20, and CYP9A19 with the highest amino acid identity of 66%. Studies of temporal expression profiles revealed that CYP9A38 showed a steady increase in mRNA level during the five instar stages, but a low-expression level in pupae, and then presented at a high-expression level again in adults. Similar expression patterns were obtained with CYP6CV1. In the fifth instar larvae, CYP6CV1 was mainly expressed in midgut and fat bodies, whereas CYP9A38 was mainly expressed in midgut. Expression studies also revealed a 3.20-fold over-expression of CYP6CV1 and 3.54-fold over-expression of CYP9A38 after larval exposure to host rice resistance. Our results suggest that both CYP6CV1 and CYP9A38 may be involved in detoxification of rice phytochemicals.     

Comparative Analyses of Cu-Zn Superoxide Dismutase (SOD1) and Thioredoxin Reductase (TrxR) at the mRNA Level between Apis mellifera L. and Apis cerana F. (Hymenoptera: Apidae) Under Stress Conditions

This study compared stress-induced expression of Cu-Zn superoxide dismutase (SOD1) and thioredoxin reductase (TrxR) genes in the European honeybee Apis mellifera L. and Asian honeybee Apis cerana F. Expression of both SOD1 and TrxR rapidly increased up to 5 h after exposure to cold (4°C) or heat (37°C) treatment and then gradually decreased, with a stronger effect induced by cold stress in A. mellifera compared with A. cerana. Injection of stress-inducing substances (methyl viologen, [MV] and H₂O₂) also increased SOD1 and TrxR expression in both A. mellifera and A. cerana, and this effect was more pronounced with MV than H₂O₂. Additionally, we heterologously expressed the A. mellifera and A. cerana SOD1 and TrxR proteins in an Escherichia coli expression system, and detection by SDS-PAGE, confirmed by Western blotting using anti-His tag antibodies, revealed bands at 16 and 60 kDa, respectively. Our results show that the expression patterns of SOD1 and TrxR differ between A. mellifera and A. cerana under conditions of low or high temperature as well as oxidative stress.     

Intracellular Immunohistochemical Detection of Tetrodotoxin in Pleurobranchaea maculata (Gastropoda) and Stylochoplana sp. (Turbellaria)

Tetrodotoxin (TTX), is a potent neurotoxin targeting sodium channels that has been identified in multiple marine and terrestrial organisms. It was recently detected in the Opisthobranch Pleurobranchaea maculata and a Platyhelminthes Stylochoplana sp. from New Zealand. Knowledge on the distribution of TTX within these organisms is important to assist in elucidating the origin and ecological role of this toxin. Intracellular micro-distribution of TTX was investigated using a monoclonal antibody-based immunoenzymatic technique. Tetrodotoxin was strongly localized in neutral mucin cells and the basement membrane of the mantle, the oocytes and follicles of the gonad tissue, and in the digestive tissue of P. maculata. The ova and pharynx were the only two structures to contain TTX in Stylochoplana sp. Using liquid chromatography-mass spectrometry, TTX was identified in the larvae and eggs, but not the gelatinous egg cases of P. maculata. Tetrodotoxin was present in egg masses of Stylochoplana sp. These data suggest that TTX has a defensive function in adult P. maculata, who then invest this in their progeny for protection. Localization in the digestive tissue of P. maculata potentially indicates a dietary source of TTX. Stylochoplana sp. may use TTX in prey capture and for the protection of offspring.     

Attack Pattern and Reproductive Ecology of Tomicus brevipilosus (Coleoptera: Curculionidae) on Pinus yunnanensis in Southwestern China

Tomicus brevipilosus (Eggers) (Coleoptera: Curculionidae, Scolytinae) was recently discovered as a new pest of Yunnan pine (Pinus yunnanensis Franchet) in Yunnan Province in southwestern China. However, little was known on its reproductive biology and pattern of trunk attack on Yunnan pine. The objectives of this study were to better understand the reproductive biology of T. brevipilosus by investigating the seasonality of trunk attacks by parent adults for the purpose of reproduction (i.e., breeding attacks) and the within-tree pattern of these attacks. Our results showed that T. brevipilosus breeding attacks in P. yunnanensis generally started in early March and ended in early June in Anning County, Yunnan. T. brevipilosus exhibited two general patterns of infestation. From early March to mid-April, T. brevipilosus bred preferentially in the trunks of Yunnan pine trees that were already infested by Tomicus yunnanensis Kirkendall and Faccoli and Tomicus minor (Hartig), colonizing spaces along the trunk (mostly in the mid- and lower trunk) that were not already occupied by the other two Tomicus species. Later, from about mid-April to early June, when there were no Yunnan pine trees newly infested by T. yunnanensis and T. minor, T. brevipilosus attacked Yunnan pine by itself, infesting the lower parts of the trunk first and then infesting progressively upward along the trunk into the crown. Infestation by T. brevipilosus extends the total period that P. yunnanensis trees are under attack by Tomicus beetles in southwestern China, which helps explain why Yunnan pine has suffered high levels of tree mortality in recent decades.     

Larvicidal Potential of the Halogenated Sesquiterpene (+)-Obtusol,Isolated from the Alga Laurencia dendroidea J. Agardh (Ceramiales: Rhodomelaceae), against the Dengue Vector Mosquito Aedes aegypti (Linnaeus) (Diptera: Culicidae)

Dengue is considered a serious public health problem in many tropical regions of the world including Brazil. At the moment, there is no viable alternative to reduce dengue infections other than controlling the insect vector, Aedes aegypti Linnaeus. In the continuing search for new sources of chemicals targeted at vector control, natural products are a promising alternative to synthetic pesticides. In our work, we investigated the toxicity of a bioactive compound extracted from the red alga Laurencia dendroidea J. Agardh. The initial results demonstrated that crude extracts, at a concentration of 5 ppm, caused pronounced mortality of second instar A. aegypti larvae. Two molecules, identified as (−)-elatol and (+)-obtusol were subsequently isolated from crude extract and further evaluated. Assays with (−)-elatol showed moderate larvicidal activity, whereas (+)-obtusol presented higher toxic activity than (−)-elatol, with a LC₅₀ value of 3.5 ppm. Histological analysis of the larvae exposed to (+)-obtusol revealed damage to the intestinal epithelium. Moreover, (+)-obtusol-treated larvae incubated with 2 µM CM-H₂DCFDA showed the presence of reactive oxygen species, leading us to suggest that epithelial damage might be related to redox imbalance. These results demonstrate the potential of (+)-obtusol as a larvicide for use against A. aegypti and the possible mode of action of this compound.     

Relative Fitness of Helicoverpa armigera (Lepidoptera: Noctuidae) on Seven Host Plants: A Perspective for IPM in Brazil

The cotton bollworm Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) is a widespread pest of many cultivated and wild plants in Europe, Africa, Asia, and Australia. In 2013, this species was reported in Brazil, attacking various host crops in the midwestern and northeastern regions of the country and is now found countrywide. Aiming to understand the effects of different host plants on the life cycle of H. armigera, we selected seven species of host plants that mature in different seasons and are commonly grown in these regions: cotton (Gossypium hirsutum, “FM993”), corn (Zea mays, “2B587”), soybean (Glycine max, “99R01”), rattlepods (Crotalaria spectabilis), millet (Pennisetum glaucum, “ADR300”), sorghum (Sorghum bicolor, “AGROMEN70G35”), and cowpea (Vigna unguiculata, “SEMPRE VERDE”). The development time of immatures, body weight, survivorship, and fecundity of H. armigera were evaluated on each host plant under laboratory conditions. The bollworms did not survive on corn, millet, or sorghum and showed very low survival rates on rattlepods. Survival rates were highest on soybean, followed by cotton and cowpea. The values for relative fitness found on soybean, cotton, cowpea, and rattlepods were 1, 0.5, 0.43, and 0.03, respectively. Survivorship, faster development time, and fecundity on soybean, cotton, and cowpea were positively correlated. Larger pupae and greater fecundity were found on soybean and cotton. The results indicated that soybean, cotton, and cowpea are the most suitable plants to support the reproduction of H. armigera in the field.     

Development of Temporal Modeling for Forecasting and Prediction of the Incidence of Lychee,Tessaratoma papillosa (Hemiptera: Tessaratomidae), Using Time-Series (ARIMA) Analysis

The most destructive enemy of the lychee, Litchi chinensis Sonn. (Sapindales: Sapindaceae), in India is a stink bug, Tessaratoma papillosa (Drury) (Hemiptera: Tessaratomidae). The population of T. papillosa on lychee trees varied from 1.43 ± 0.501 to 9.85 ± 3.924 insects per branch in this study. An increase in the temperature and a decrease in the relative humidity during summer months (April to July) favor the population buildup of T. papillosa. A forecasting model to predict T. papillosa incidences in lychee orchards was developed using the autoregressive integrated moving average (ARIMA) model of time-series analysis. The best-fit model for the T. papillosa incidence was ARIMA (1,1), where the P-value was significant at 0.01. The highest T. papillosa incidences were predicted for April in 2010, January in 2011, May in 2012, and February in 2013. A model based on time series offers longer-term forecasting. The forecasting model, ARIMA (1,1), developed in this study will predict T. papillosa incidences in advance, thus providing functional guidelines for effective planning of timely prevention and control measures.     

Flagellar Sensillar Equipment of Two Morphologically Closely Related Aphid Hyperparasitoids (Hymenoptera: Figitidae: Alloxysta)

The antennal sensillar equipment in the parasitic wasp family Figitidae was analyzed to date only in few species, despite some are associated with crop pests and can have an economic importance. It is the case of the genus Alloxysta, which includes hyperparasitoids of aphids which can potentially reduce effectiveness of primary pest parasitoids. Here we analyzed, through scanning electron microscopy, the diversity, morphology, and distribution of the antennal sensilla in males and females of Alloxysta consobrina (Zetterstedt) and Alloxysta victrix (Westwood), two species with overall very similar morphology. In both species, antennae are filiform and cylindrical, and flagellum was longer in A. victrix. Eight sensillar types have been recognized: four types of sensilla trichoidea (ST-A, ST-B, ST-C, ST-D), sensilla coeloconica, sensilla placoidea, sensilla campaniformia, and sensilla basiconica. ST-A, ST-B, ST-C, and sensilla placoidea were the most abundant types on the antennae and often increased in number and decreased in size toward the tip of antenna. The two species seem to have several differences in their sensillar equipment, possibly in accordance with the different degree of host range. On the other hand, sexual dimorphism is probably due to the different stimuli that have to be correctly processed. The comparison with the other species of Figitidae studied by far showed, at subfamily-level, that variability in sensillar equipment and phylogeny do not agree. This suggests a complex series of morphological changes during evolution of this group. The taxonomic sample should be thus substantially enlarged to disclose possible trends in sensillar equipment evolution in the family.     

Prevalence of Endosymbionts in Polish Populations of Leptinotarsa decemlineata (Coleoptera: Chrysomelidae)

Colorado potato beetle (CPB, Leptinotarsa decemlineata Say) (Coleoptera: Chrysomelidae) is one of the most serious insect pest feeding on wild and cultivated Solanaceae plants. This pest poses a significant threat to potato crops. CPB originated from North America but has become widespread and has adapted in new localizations. Currently, it is reported in many countries worldwide. Endosymbiotic bacteria might have an influence on insect adaptation to new conditions. They are known to play a role in invasiveness of insect hosts and to facilitate colonization of new niches; however, information on endosymbionts of the CPB is very limited. In this study, we screened CPB populations collected from 20 evenly distributed locations in Poland for the presence of Arsenophonus, Cardinium, Wolbachia, and Flavobacterium. We found the presence of Flavobacterium in the studied insects. Little is known about CPB–endosymbionts interactions, thus this study may provide a reference for future studies in this subject.     

Chemical Composition,Antifeedant, Repellent,and Toxicity Activities of the Rhizomes of Galangal,Alpinia galanga Against Asian Subterranean Termites,Coptotermes gestroi and Coptotermes curvignathus (Isoptera: Rhinotermitidae)

Dual choice bioassays were used to evaluate the antifeedant property of essential oil and methanolic extract of Alpinia galanga (L.) (locally known as lengkuas) against two species of termites, Coptotermes gestroi (Wasmann) and Coptotermes curvignathus (Holmgren) (Isoptera: Rhinotermitidae). A 4-cm-diameter paper disc treated with A. galanga essential oil and another treated with either methanol or hexane as control were placed in a petri dish with 10 termites. Mean consumption of paper discs (miligram) treated with 2,000 ppm of essential oil by C. gestroi was 3.30 ± 0.24 mg and by C. curvignathus was 3.32 ± 0.24 mg. A. galanga essential oil showed significant difference in antifeedant effect, 2,000 ppm of A. galanga essential oil was considered to be the optimum concentration that gave maximum antifeedant effect. The essential oil composition was determined using gas chromatography-mass spectrometry. The major component of the essential oil was 1,8-cineol (61.9%). Antifeedant bioassay using 500 ppm of 1,8-cineol showed significant reduction in paper consumption by both termite species. Thus, the bioactive agent in A. galangal essential oil causing antifeeding activity was identified as 1,8-cineol. Repellent activity shows that 250 ppm of 1,8-cineol caused 50.00 ± 4.47% repellency for C. gestroi, whereas for C. curvignathus 750 ppm of 1,8-cineol was needed to cause similar repellent activity (56.67 ± 3.33%). C. curvignathus is more susceptible compare to C. gestroi in Contact Toxicity study, the lethal dose (LD₅₀) of C. curvignathus was 945 mg/kg, whereas LD₅₀ value for C. gestroi was 1,102 mg/kg. Hence 1,8-cineol may be developed as an alternative control against termite in sustainable agriculture practices.