Male and female Popillia quadriguttata (Fabricius) and Protaetia brevitarsis (Lewis) (Coleoptera: Scarabaeidae) response to Japanese beetle floral and pheromone lures

Popillia quadriguttata (Fabricius), and Protaetia brevitarsis (Lewis) adults were captured with Japanese beetle, Popillia japonica Newman, sex attractant and floral lures at Changchun, China during July–August 2012. The floral lure (phenethyl propionate:eugenol:geraniol, 3:7:3) was attractive to male and female P. quadriguttata (AV: 1.2 ± 0.9; 1.1 ± 0.3; total: 2.3 ± 0.8), and was similar in attraction to the combination of the sex attractant (SA) [(R, Z)-5-(1-decenyl) dihydro-2(3H)-furanone] plus the floral lure for male (1.60 ± 0.2), female (1.30 ± 1.1) and total captures (2.9 ± 3.0). However, the SA alone captured only males in much higher numbers than when combined with the floral lure (10.0 ± 6.4). In a separate earlier test, the greatest number of P. quadriguttata males (12.5 ± 3.0), female (12.2 ± 1.5) and total captures (24.7 ± 2.5) was in yellow, laboratory-made, bottle traps. The floral lure also attracted female Pro. brevitarsis (10.0 ± 3.4), while the SA attracted only few male beetles (1.0 ± 0.2). The combination SA + floral lure captured similar females (11.0 ± 2.0) and total (14.2 ± 2.2) Pro. brevitarsis as the floral lure alone. Two butterflies, Colias erate poliographus (Motschulsky) and Pieris rapae (Linnaeus), were also attracted to the floral lure. These studies indicate a potential for replacing pesticides by using the Japanese beetle lures for monitoring and control of several insects in China, and that they would be useful in monitoring and eradication of two potential scarab pests, P. quadriguttata and Pro. brevitaris, in the United States and Europe.     

Attraction of the predator,Harmonia axyridis (Coleoptera: Coccinellidae), to its prey,Myzus persicae (Hemiptera: Aphididae), feeding on Chinese cabbage

The predatory multicolored Asian lady beetle, Harmonia axyridis, was attracted to volatiles released from Chinese cabbage infested by the green peach aphid, Myzus persicae, in T-tube olfactometer choice tests. However, lady beetle adults and larvae did not respond to clean air, Chinese cabbage alone or green peach aphid alone. Of different prey densities, H. axyridis adults were most attracted to Chinese cabbage infested by 60 M. persicae adults after 24 h. However, H. axyridis larvae were not attracted to Chinese cabbage infested by M. persicae. Mechanically damaged Chinese cabbage attracted neither lady beetle adults nor larvae. Predatory adults were attracted to 60 M. persicae adults after 24 and 48 h, and to 90 M. persicae adults after 12 h, suggesting that the predatory response depends on the prey density. Lady adult beetles did not prefer the volatiles induced by Diamondback moth, Plutella xylostella, indicating that specific host insect specificity attracts respective natural enemies. It can be explained that the volatile compounds emitted from the host plant as a result of herbivore attack preferred by the specific insect species.     

Attraction of garden chafer, Phyllopertha horticola, to floral Japanese beetle lure

Field experiments using funnel traps were performed on a golf course to investigate the attractiveness of a floral Japanese beetle lure for adults of the garden chafer, Phyllopertha horticola, a common European turf pest. Traps baited with 10 mg of the floral lure consisting of phenylethyl propionate + eugenol + geraniol (3 : 7 : 3) captured more adult garden chafers of both sexes than non‐baited control traps. In contrast, 1,3‐dimethyl‐2,4‐(1H, 3H)‐quinazolinedione, the sex pheromone of the closely related pale‐brown chafer P. diversa, neither influenced alone nor in combination with the floral lure the behaviour of adult garden chafers. The results suggest that the floral Japanese beetle lure might be used for monitoring or control of P. horticola.     

Seasonal Changes in Habitat Preference by Coleomegilla maculata: Implications for Colorado Potato Beetle Management in Potato

Seasonal abundance, dispersal, and overwintering of the 12-spotted ladybird beetle, Coleomegilla maculata (DeGeer), in the mid-Atlantic states were investigated to assess the potential to manipulate the habitat of this predator to increase its impact on Colorado potato beetle, Leptinotarsa decemlineata (Say), management in commercial potato plantings. C. maculata populations were highest in corn fields through August, and overwintering aggregations were found most commonly in habitats adjacent to these fields. Although adult and larval populations of C. maculata were synchronized with first-generation potato beetle eggs and small larvae, they were concentrated in wheat rather than potato fields because most of the potato plants had not yet emerged when C. maculata adults dispersed from their overwintering sites. The subsequent generation of C. maculata tended to aggregate in corn rather than potato in late May and early June. Therefore, given the current mixture of crops as well as climatic and marketing limitations on planting date for potatoes, the potential for habitat manipulation to increase the impact of C. maculata on Colorado potato beetle management in potato in the mid-Atlantic states appears to be limited.     

Diet selection and eating behaviour of lactating goats subjected to time restricted feeding in choice and single feeding system

This study was carried out to investigate diet selection and eating behaviour of lactating German Fawn × Hair Crossbred goats in different feeding methods and levels. Twenty German Fawn × Hair first backcross does (B₁) were allocated into 4 treatment groups (2 feeding methods single (TMR) and choice feeding × 2 feeding levels ad libitum and restricted) with 5 replicates. Restricted feeding was applied only 4 h feed allocation during day. Barley, corn, soybean meal, corn gluten meal, wheat bran and alfalfa hay were feed ingredients for single and choice feeding. Eating patterns, milk yield and composition were determined for 8 weeks. The following results were obtained: (1) the meal criteria for goats restricted single and choice-fed, ad libitum single and choice-fed were determined as 1.00 and 0.63, 12.88 and 10.23 min, respectively. (2) Ad libitum feeding increased meal size, meal length, intermeal interval, total eating duration and decreased eating rate and meal number, compared to restricted feeding (P < 0.01). Choice feeding decreased meal size (P < 0.05), meal length (P < 0.01) and increased eating rate and meal number (P < 0.01), compared to single feeding. Restricted fed goats decreased intermeal interval in single feeding compared to choice feeding (P < 0.01), but increased meal number in choice feeding (P < 0.01). (3) Ad libitum choice-fed does made a diet containing 12.79% corn, 35.41% barley, 13.21% wheat bran, 5.35% soybean meal, 1.28% corn gluten meal and 29.80% alfalfa meal while restricted choice-fed does made a diet having more corn (27.69%), corn gluten meal (5.62%) and wheat bran (16.17%) and less barley (14.37%) and soybean meal (4.51%). (4) Choice feeding decreased RUP intake (P < 0.05) without affecting milk protein, irrespective to feeding levels, while having a tendency to increase in milk yield (14.2%) and 4% FCM (8.8%). (5) Restricted feeding decreased DM, ME, ADF and NDF intakes (P < 0.05) with concomitant decreases in 4% FCM, total milk solid, ash and fat compositions (P < 0.05), irrespective to feeding methods. (6) Choice-fed goats changed their preferences for a possible synchronized nutrient intake during a daytime, as sorted barley, soybean meal and alfalfa hay from early morning to late afternoon.It could be concluded that choice-fed goats have the ability to make their diet to meet nutrient requirements and had a tendency to increase in milk yield. Restriction in feeding time resulted in lower feed intake and milk yield, although the animal changed their feed preference in favour of high quality ingredients and eating pattern with lower meal criterion and intermeal interval.     

Determination of coleopteran insects associated with spore dispersal of Cryptoporus volvatus (Polyporaceae: Basidiomycota) in Korea

The veiled polypore, Cryptoporus volvatus, is distributed widely in North America and East Asia and is believed to have a mutualistic relationship with coleopteran species—the fungus providing food and shelter in basidiocarps and beetle helping disperse spores.Seventy fresh basidiocarps of C. volvatus were collected from the Japanese red pine (Pinus densiflora) in the spring season of 2013 from two sites in Korea. A total of 251 insects (101 adult and 150 larvae) were collected from the inside of basidiocarps and identified using morphology and mitochondrial cytochrome c oxidase subunit I (COI) sequences. Six species belonging to five coleopteran families were identified. The number of spores attached to the bodies of adult insects was counted and average spore counts for each of the six species ranged between 1.0 × 10⁴ and 5.2 × 10⁵ spores/individual. Across localities, three species were shared (Aethina suturalis, Trogossita japonica and Parabolitophagus felix) and carried spores at high densities on their bodies, making them more likely to aid in spore dispersal.     

Life cycle of Anthracophora rusticola (Coleoptera: Cetoniidae) from the commensal nests of Chinese Sparrowhawks (Accipiter soloensis)

Commensal breeding habits of the Anthracophora rusticola (Coleoptera: Cetoniidae) in stick nests of carnivorous, piscivorous, and omnivorous birds were recently reported, but the detailed life history of the scarab beetle has not been thoroughly elucidated. To understand the breeding cycle of the declining A. rusticola related to that of nesting Chinese sparrowhawks (Accipiter soloensis) in Korea, we examined six sparrowhawk nests from 2005 to 2008 and reared larvae and cocoons in a cage in 2008. Each sparrowhawk nest hosted 11.3 ± 3.0 larvae and cocoons, implying that sparrowhawk nests might be the most important nesting resource of the beetles in Korea. The beetles may lay their eggs between late May and early July after nest building by sparrowhawks. Larvae were observed from July to August in the sparrowhawk nests, and adult beetles emerged in September after a mean pupal period of 25.5 days. Our results on the coinciding breeding of two commensal taxa over three months from May to September may provide key information regarding the life history of the commensal scarabid beetle and potential causes of the concurrent decline of both taxa in Korea.     

Distribution of Sunn Pest,Eurygaster integriceps Puton (Hemiptera: Scutelleridae), in overwintering sites

Sunn Pest, Eurygaster integriceps Puton (Hemiptera: Scutelleridae), is the most serious pest of wheat in West and Central Asia. Following harvest, the pest migrates from wheat fields to high elevations to escape high summer temperatures and, the following spring, moves back to the fields. In this work, Sunn Pest populations were monitored in 21 overwintering sites located in seven provinces (with 800–2500 m elevation) in Iran. Litter samples were taken twice in each site, once in July–November 2002 and again in March 2003. At each site 60 Artemesia sp. bushes were selected at random and litter from each bush was collected and the number of live male and female adult Sunn Pest counted. More Sunn Pests were collected in higher elevations in the first sampling. Golestan (893 m), Kermanshah (1583 m), and Qazvin (1675 m), located in valleys and surrounded by mountainous areas, had < 5 adults/bush. However Azarbayejan (1750 m), Markazi (1883 m), Kordestan (2067 m), and Esfahan (2141 m), located in mountainous areas and/or close to mountains, had 10–52 adults/bush. No noticeable changes in Sunn Pest populations were observed in 17 overwintering sites among 21 between the two sampling times. These results suggest that, after Sunn Pest moves to more highly elevated sites at aestivation time, most of them remain in these sites during overwintering before migration to wheat fields in the spring. Sunn Pest can be targeted for management in these areas during the fall and/or early spring as their movement is clearly limited.     

Attractiveness to Anastrepha ludens (Diptera: Tephritidae) of plant essential oils and a synthetic food-odour lure

Abstract:  Experiments were conducted in a citrus orchard to investigate the attractiveness of 26 plant essential oils individually and in combination with a synthetic food odour lure to the Mexican fruit fly, Anastrepha ludens Loew. Anise, rose/grape seed, and tea tree oils were more attractive than unbaited traps but none approached the attractiveness of Advanced Pheromone Technologies' AFF lure, a synthetic food-odour lure that emits several nitrogenous chemicals attractive to this fly. Traps baited with most of the oils were less attractive than unbaited traps. Rose/grape seed oil and pure-rose oil enhanced attractiveness of AFF lures to both males and females by about 68%. Grape seed oil did not enhance the attractiveness of AFF lures demonstrating that rose oil was the active component of the rose/grape seed oil. No other oil enhanced attractiveness of AFF lures and most decreased attraction to AFF lures. The possibility that highly attractive chemicals may be present in rose oil as minor components is discussed. Traps baited with the combination of clove bud oil and the AFF lure captured only 3% as many flies as traps baited only with the AFF lure indicating that clove bud oil is highly repellent to Mexican fruit flies.     

Selective flowers to enhance biological control of cabbage pests by parasitoids

Habitat management is an important element in sustainable agriculture and can be used to maximize a range of ecosystem services that support crop production. An important example of such ecosystem services is biological control of pests which can be enhanced by providing arthropod natural enemies with suitable floral resources. The potential risk of this approach, however, is that flowering plants may enhance the fitness of the targeted pests as well. We conducted experiments to identify selective plant species that would improve the longevity and parasitization rate of the parasitoid wasp Microplitis mediator without benefiting its host pest, the cabbage moth Mamestra brassicae. Effects on longevity were also assessed for Diadegma fenestrale, a generalist parasitoid wasp attacking lepidopteran pests. Additionally, we compared the effects of floral and extrafloral nectar, the latter being formed in some plant species and can significantly prolong the duration of nectar availability for natural enemies. Longevity of M. mediator and D. fenestrale as well as parasitization rates of M. mediator were significantly increased by the presence of Fagopyrum esculentum (floral nectar), Centaurea cyanus (floral and extrafloral nectar) and non-flowering Vicia sativa (extrafloral nectar). M. mediator parasitized 202.3 ± 29.7 M. brassicae larvae during its lifetime when presented F. esculentum, compared to 14.4 ± 3.4 larvae in the absence of floral resources. Extrafloral nectar of C. cyanus and V. sativa was as suitable for M. mediator as floral nectar and significantly increased longevity and parasitization rates. Longevity and fecundity of M. brassicae were not supported by the plant species tested. These results stress the importance of plant screening to achieve plant selectivity and to maximize biological control. F. esculentum, C. cyanus and V. sativa are recommended as selective plant species to enhance parasitoids of M. brassicae.     

A freeze-dried diet to test pathogens of Colorado potato beetle

The Colorado potato beetle is an important pest on potato, eggplant, and tomato. Because Colorado potato beetles develop resistance to insecticides quickly, new methods are needed for control. Bacillus thuringiensis is the only bacterium to successfully control Colorado potato beetle. Until recently, one of the drawbacks to testing bacteria against the Colorado potato beetle has been the lack of an artificial diet for screening. Previous artificial diets will only be consumed by Colorado potato beetle larvae when fresh. To improve storage, we developed a freeze-dried diet, based on a 96-well plate, suitable to feed larvae for the duration of a bioassay. Individual diet components were tested both for their effect on insect growth and on pathogen toxicity. When the preservatives, methylparaben and sorbic acid, were removed from the diet, the average weight of second instar larvae increased from 7.9 mg to greater than 9.8 mg. The preservatives inhibited the growth of two of the bacteria tested, Photorhabdus luminescens HM and Chromobacterium sp. PRAA. The removal of these preservatives also allowed for fungal growth and reduced survival from 94 to 38%. Removing diet preservatives, that inhibited the growth of Chromobacterium sp. PRAA, increased the total mortality of the larvae as well as reducing the time needed to kill 50% of the larvae. Compared to incorporation of bacteria into molten diet, the total mortality of Colorado potato beetle fed either P. luminescens HM or Chromobacterium sp. PRAA on freeze-dried diet doubled. Preparation of freeze-dried diet need not be synchronized with the insect or the pathogen. The freeze-dried diet gave consistent results as measured by low control mortality and pathogen toxicity over time.     

Interactions between Spathius agrili (Hymenoptera: Braconidae) and Tetrastichus planipennisi (Hymenoptera: Eulophidae), larval parasitoids of Agrilus planipennis (Coleoptera: Buprestidae)

Three hymenopteran parasitoids native to China are being released in the United States as biological control agents for the emerald ash borer (EAB), Agrilus planipennis Fairmaire, an Asian buprestid species responsible for mortality of ash trees (Fraxinus spp.) in North America. Two of these hymenopterans, Spathius agrili Yang (Braconidae), a larval ectoparasitoid, and Tetrastichus planipennisi Yang (Eulophidae), a larval endoparasitoid, prefer late-instar EAB larvae. This overlapping host preference raises concerns that interspecific competition following field releases may compromise establishment of one or both species. In a series of laboratory and field experiments, we found S. agrili and T. planipennisi exhibited similar parasitism rates when presented alone with EAB larvae for 12–14 days. However, S. agrili was more efficient at locating and parasitizing hosts within the first 27 h, possibly explaining why S. agrili excluded T. planipennisi in the laboratory trials and nearly excluded T. planipennisi in field trials when the two species were presented together with EAB larvae. We found that S. agrili parasitized larvae previously parasitized by T. planipennisi but not the reverse. However, S. agrili offspring failed to complete development on hosts that were previously parasitized by T. planipennisi. We recommend releasing these species separately in time or space to avoid the antagonistic interactions observed in this study.     

Cold tolerance of the maize orange leafhopper,Cicadulina bipunctata

Cicadulina bipunctata was originally distributed in tropical and subtropical regions of the Old World. This leafhopper recently expanded its distribution area to southern parts of temperate Japan. In this study, factors affecting the overwintering ability of C. bipunctata were examined. A series of laboratory experiments revealed that cold acclimation at 15 °C for 7 days enhanced the cold tolerance of C. bipunctata to the same level as an overwintering population, adult females were more tolerant of cold temperature than adult males, and survival of acclimated adult females was highly dependent on temperature from −5 to 5 °C and exposure duration to the temperature. The temperature of crystallization of adult females was approximately −19 °C but temperatures in southern temperate Japan rarely dropped below −10 °C in the winter, indicating that overwintering C. bipunctata adults in temperate Japan are not killed by freezing injury but by indirect chilling injury caused by long-term exposure to moderately low temperatures. An overwintering generation of C. bipunctata had extremely low overwinter survival (<1%) in temperate Japan; however, based on winter temperature ranges, there are additional areas amenable to expansion of C. bipunctata in temperate Japan.     

An adjustable action threshold using larval parasitism of Helicoverpa armigera (Lepidoptera: Noctuidae) in IPM for processing tomatoes

An adjustable action threshold that uses estimates of larval parasitism of Helicoverpa armigera (Hübner) in individual fields was assessed over three consecutive years in processing tomatoes in Hawke’s Bay, New Zealand. Tomato fields were monitored weekly for levels of infestation by H. armigera larvae. When infestation levels became of concern, either approaching or exceeding the standard action threshold of one larva per plant, collections of 30+ representative larvae were made. These larvae were measured and reared individually at 30 °C, and after 4 days rates of parasitism were estimated visually. From these data, spray recommendations were made using a formula that adjusted the action threshold to allow for mortality from parasitism. At harvest, damage assessments were made to validate these recommendations. Results showed that estimates of parasitism after 4 days were accurate predictors of final assessments of parasitism. Overall parasitism during the three seasons was 71%, confirming that the original threshold, which relies on about 50% parasitism, needed revision. The dominant parasitoid was Cotesia kazak, reared from 91% of all parasitized larvae over the 3 years. Microplitis croceipes and the self introduced polyphagous parasitoid Meteorus pulchricornis were reared from 4% and 5% of the parasitized larvae respectively. In 16 of 17 fields examined, the adjustable threshold kept fruit damage at harvest below the tolerated level of 5%. The single crop with excessive damage had only 0.5% fruit damage above this level. This adjustable threshold, which varied in this study from 1–8.3 larvae per plant, has been incorporated into an updated IPM programme and contributed to a 95% reduction in insecticide use.     

Horseradish peroxidase production from Spodoptera frugiperda larvae: A simple and inexpensive method

Horseradish peroxidase is used in many biotechnological fields including diagnostics, biocatalysts and biosensors. Horseradish peroxidase isozyme C (HRPC) was extracellularly expressed in Spodoptera frugiperda Sf9 cell culture and in intact larvae. At day 6 post-infection, the concentration of active HRPC in suspension cultures was 3.0 ± 0.1 μg per 1 × 10⁶ cells or 3.0 ± 0.1 mg l⁻¹ with a multiplicity of infection of 1 in the presence of 7.2 μM hemin. Similar yields were obtained in monolayer cultures. In larvae, the HRPC expression level was 137 ± 17 mg HRPC kg⁻¹ larvae at day 6 post-infection with a single larvae thus producing approximately 41 μg HRPC. The whole larval extract was separated by ion exchange chromatography and HRPC was purified in a single step with a yield of 75% and a purification factor of 117. The molecular weight of recombinant HRPC was 44,016 Da, and its glycosylation pattern agreed with that expected for invertebrates. The K_m and V_max were 12.1 ± 1.7 mM and 2673 ± 113 U mg⁻¹, respectively, similar to those of HRP purified from Armoracia rusticana roots. The method described in this study, based on overexpression of HRPC in S. frugiperda larvae, is a simple and inexpensive way to obtain high levels of active enzyme for research and other biotechnological applications.     

Effect of the interaction between Anticarsia gemmatalis multiple nucleopolyhedrovirus and Epinotia aporema granulovirus,on A. gemmatalis (Lepidoptera: Noctuidae) larvae

The bean shoot borer Epinotia aporema Wals. (Lepidoptera: Tortricidae) and the velvet bean caterpillar Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae) are key pests of soybean and other legume crops in South America. They are often found simultaneously in certain regions. A. gemmatalis nucleopolyhedrovirus (AgMNPV) is widely used to control A. gemmatalis. More recently, E. aporema granulovirus (EpapGV) has been characterized and evaluated as a bioinsecticide for E. aporema. In order to increase its potential use and to design optimized strategies for the management of lepidopteran pests, we evaluated the interaction between EpapGV and AgMNPV on third instar A. gemmatalis larvae. Larvae fed with 50 AgMNPV OBs/larva showed an increase in the mortality rates (from 42% to 81%) and a decrease in the median survival time (from 7.7 days to 5.7 days) when these OBs were mixed with 6000 EpapGV OBs/larva. When 300 AgMNPV OBs/larva were used alone or in combination with EpapGV OBs no changes in biological parameters were observed. No mortality was detected in A. gemmatalis larvae treated with EpapGV alone. In larvae fed with the viral mixtures, only AgMNPV DNA was detected by PCR. A. gemmatalis peritrophic membranes (PMs) examined by SDS–PAGE and scanning electron microscopy showed signs of damage. Notably, we found the presence of spheroidal bodies associated with damaged areas in the PMs of larvae fed with EpapGV but not in those that were given AgMNPV alone. These results show that EpapGV increases the viral potency of AgMNPV, and thus the insecticidal efficiency, suggesting that the use of formulations including both viruses might be a valuable tool for pest management.     

Lethal effects of ichthyotoxic raphidophytes,Chattonella marina,C. antiqua,and Heterosigma akashiwo,on post-embryonic stages of the Japanese pearl oyster,Pinctada fucata martensii

The inimical effects of the ichthyotoxic harmful algal bloom (HAB)-forming raphidophytes Heterosigma akashiwo, Chattonella marina, and Chattonella antiqua on the early-life stages of the Japanese pearl oyster Pinctada fucata martensii were studied. Fertilized eggs and developing embryos were not affected following exposure to the harmful raphidophytes; however, all three algal species severely affected trochophores and D-larvae, early-stage D-larvae, and late-stage pre-settling larvae. Exposure to C. marina (5 × 10² cells ml⁻¹), C. antiqua (10³ cells ml⁻¹), and H. akashiwo (5 × 10³ cells ml⁻¹) resulted in decreased success of metamorphosis to the trochophore stage. A complete inhibition of trochophore metamorphosis was observed following exposure to C. antiqua at 5 × 10³ cells ml⁻¹ and C. marina at 8 × 10³ cells ml⁻¹. In all experiments, more than 80% of newly formed trochophores were anomalous, and in the case of exposure to H. akashiwo at 10⁵ cells ml⁻¹ more than 70% of D-larvae were anomalous. The activity rates of D-larvae (1-day-old) were significantly reduced following exposure to C. antiqua (8 × 10³ cells ml⁻¹, 24 h), C. marina (8 × 10³ cells ml⁻¹, 24 h), and H. akashiwo (10⁴ cells ml⁻¹, 24 h). The activity rates of pre-settling larvae (21-day-old) were also significantly reduced following exposure to C. antiqua (10³ cells ml⁻¹, 24 h), C. marina (8 × 10³ cells ml⁻¹, 24 h), and H. akashiwo (5 × 10⁴ cells ml⁻¹, 24 h). Significant mortalities of both larval stages were induced by all three raphidophytes, with higher mortality rates registered for pre-settling larvae than D-larvae, especially following exposure to C. marina (5 × 10²–8 × 10³ cells ml⁻¹, 48–86 h) and C. antiqua (10³–8 × 10³ cells ml⁻¹, 72–86 h). Contact between raphidophyte cells and newly metamorphosed trochophores and D-larvae, 1-day-old D-larvae, and 21-day-old larvae resulted in microscopic changes in the raphidophytes, and then, in the motile early-life stages of pearl oysters. Upon contact and physical disturbance of their cells by larval cilia, H. akashiwo, C. marina and C. antiqua became immotile and shed their glycocalyx. The trochophores and larvae were observed trapped in a conglomerate of glycocalyx and mucus, most probably a mixture of larval mucous and raphidophyte tricosyts and mucocytes. All motile stages of pearl oyster larvae showed a typical escape behavior translating into increased swimming in an effort to release themselves from the sticky mucous traps. The larvae subsequently became exhausted, entrapped in more heavy mucous, lost their larval cilia, sank, become immotile, and died. Although other toxic mediators could have been involved, the results of the present study indicate that all three raphidophytes were harmful only for motile stages of pearl oysters, and that the physical disturbance of their cells upon contact with the ciliary structures of pearl oyster larvae initiated the harmful mechanism. The present study is the first report of lethal effects of harmful Chattonella spp. towards larvae of a bivalve mollusc. Blooms of H. akashiwo, C. antiqua and C. marina occur in all major cultivation areas of P. fucata martensii during the developmental period of their larvae. Therefore, exposure of the motile early-life stages of Japanese pearl oysters could adversely affect their population recruitment. In addition, the present study shows that further research with early-life development of pearl oysters and other bivalves could contribute to improving the understanding of the controversial harmful mechanisms of raphidophytes in marine organisms.     

Evaluation of distant carbon sources in biosurfactant production by a gamma ray-induced Pseudomonas putida mutant

Pseudomonas putida 33 wild strain, subjected to gamma ray mutagenesis and designated as P. putida 300-B mutant was used as microbial rhamnolipid-producer by using distant carbon sources (viz. hydrocarbons, waste frying oils ‘WFOs’, vegetable oil refinery wastes and molasses) in the minimal media under shake flask conditions. The behavior of glucose as co-substrate and growth initiator was examined. The 300-B mutant strain showed its ability to grow on all the substrates tested and produced rhamnolipid surfactants to different extents however; soybean and corn WFOs were observed to be preferred carbon sources followed by kerosene and paraffin oils, respectively. The best cell biomass (3.5 g l⁻¹) and rhamnolipids yield (4.1 g l⁻¹) were obtained with soybean WFO as carbon source and glucose as growth initiator under fed-batch cultivation showing an optimum specific growth rate (μ) of 0.272 h⁻¹, specific product yield (q_p) of 0.318 g g⁻¹ h and volumetric productivity (P_V) of 0.024 g l⁻¹ h. The critical micelle concentration of its culture supernatant was observed to be 91 mg rhamnolipids l⁻¹ and surface tension as 31.2 mN m⁻¹.     

Recent advances in biological control of important native and invasive forest pests in China

In recent decades, China has suffered severe attacks by both native and invasive forest pests. We have carried out a series of research projects on biological control of these pests. The fall webworm, Hyphantria cunea (Drury) (Lepidoptera: Arctiidae) has been sustainably controlled by an effective gregarious pupal endoparasitoid, Chouioia cunea Yang (Chalcidoidea: Eulophidae), which is native to China, and spraying HcNPV virus against the pest’s larval stage. Pine wilt disease, caused by the pine wood nematode (Bursaphelenchus xylophilus (Steiner et Buhrer) Nickle) (Aphelenchida: Aphelenchidae), is currently the number one pest in China. The strategy for controlling the disease is to manipulate the nematode’s vector, Monochamus alternatus Hope (Coleoptera: Cerambycidae). We discovered that Dastarcus helophoroides (Fairmaire) (Coleoptera: Bothrideridae) is the most important natural enemy in China pine forests. Mass rearing and release techniques were studied and developed. By releasing the parasitoid, 92.6% of the M. alternatus were parasitized in the first year. Meanwhile, three elaterid beetle species were found to prey on the larva of M. alternatus. The red turpentine beetle, Dendroctonus valens (LeConte) (Coleoptera:Scolytidae) was suppressed by a predator, Rhizophagus grandis Gyllenhal (Coleoptera: Rhizophagidae) introduced from Belgium and a total 3334 ha. of pine forests were protected. The oak longhorned beetle, Massicus raddei (Blessig) (Coleoptera: Cerambycidae) is the number one pest in the northeast forests of China, where it damages trunk of oaks, mainly Quercus liaotungensis and Q. mongolicus. An integrated management technique was developed for controlling the longhorned beetle: a special black light was invented for trapping the adults; the parasitoid Sclerodermus pupariae Yang et Yao (Hymenoptera: Bethylidae) was released against young larvae; and the parasitoid Dastarcus helophoroides eggs and/or adults were released when the hosts were mature larvae and/or pupae. By applying the technique for five years in northeastern China oak forests, the oak longhorned beetle has been controlled to a large extent. The emerald ash borer, Agrilus planipennis Fairmaire (Coleoptera: Buprestidae), is native to eastern Asia, including China, and feeding by larvae damages ash trees. Natural enemies of the emerald ash borer were investigated and seven species were found in China, of which Spathius agrili Yang (Hymenoptera: Braconidae), Tetrastichus planipennisi Yang (Hymenoptera: Eulophiae), Sclerodermus pupariae Yang et Yao and Oobius agrili Zhang et Huang (Hymenoptera: Encyrtidae) are predominant and have high potential for biocontrol of the pest. The biology, behavior, ecology and mass rearing techniques of the parasitoids were studied.     

Natural parasitism of Romanomermis iyengari (Welch) (Nematoda: Mermithidae) on various species of mosquitoes breeding in rice fields in Pondicherry,India

Natural habitat of Romanomermis iyengari, an entomophilic mermithid nematode, is rice field. Parasitism of this nematode on mosquito larvae breeding in rice fields in Pondicherry, India, has been studied in 1979. The present study was to find out its persistence in the same habitat, after many years of agricultural practices. Evaluation of parasitism on mosquito larvae was carried out during two rice cultivation seasons at a schedule of 1 week prior, and 1 and 2 weeks after transplantation of paddy. Larval stages of mosquito species such as Culex tritaeniorhynchus, Culex vishnui, Anopheles vagus, and Anopheles subpictus showed parasitism of R. iyengari. Parasitism was the highest on C. tritaeniorhynchus with a maximum of 15.38% during season I and 17.85% during season II. Followed by this, it was on A. vagus (10.82% and 9.43%), C. vishnui (7.37% and 8.69%), and A. subpictus (3.70% and 6.36%). The overall density of mosquito larvae and level of parasitism of R. iyengari on them during season II was significantly higher than that of season I. The parasitism was maximum at 1 week before transplantation and showed a decreasing trend towards 2 weeks after transplantation, during both the seasons. The level of natural parasitism of R. iyengari during the present investigation was higher than that observed in the same ecosystem almost 35 years ago. This is clearly an indication towards the capability of the nematode to withstand several years of agricultural practices, including pesticide and agrichemical application. Hence, if applied at sufficient level and frequency, this nematode could act as a successful biocontrol agent against mosquitoes breeding in rice fields.