Notes on the morphology,ecology and life cycles of Fukaurahydra anthoformis and Hataia parva (Hydrozoa,Athecata)

Fukaurahydra anthoformis and Hataia parva are solitary athecate hydroids occurring in northern Japan. New information on the external morphology, nematocysts, ecology, and life cycles of these species is presented. It is noteworthy that H. parva bears stenoteles, which are generally not found among the families of Filifera. Neither species produces free medusae. The eggs are fertilized in the female gonophores, from which unciliated larvae are released. These larvae do not swim and soon attach to a substrate. After attachment the larvae become covered by a sheath to form cysts. The cysts rest on a substrate without any outer change for several months. As the water temperature drops in autumn to early winter the cysts begin to hatch, forming tiny polyps after the larva creeps out from the chitinous sheath. Cyst formation proves to be common also in other solitary hydroids, most of which are inhabitants of cool or cold waters.     

Effect of pollen,natural prey and factitious prey on the development of <Emphasis Type="Italic">Iphiseius degenerans</Emphasis>

The predatory mite Iphiseius degenerans (Berlese) is commercially available as a biological control agent of thrips and spider mites in greenhouse crops. Developmental duration and immature survival of I. degeneransreared on nine types of food (almond pollen, apple pollen, castor bean pollen, plum pollen, sweet pepper pollen, Tetranychus urticaeKoch, Frankliniella occidentalis(Pergande), Ephestia kuehniella Zeller eggs and Artemia franciscana Kellogg cysts) and on three substrates (Multicel, sweet pepper leaf, and bean leaf) were determined in the laboratory. All experiments were carried out at 25 °C. On Multicel, mean developmental times on pollen ranged from 6.0 to 7.1 days, with the lowest value recorded on almond pollen and the highest on sweet pepper pollen. When reared on castor bean pollen significantly longer developmental times were obtained on a sweet pepper leaf compared to a bean leaf or Multicel. Developmental duration when offered T. urticaeon Multicel ranged between 6.1 and 6.9 days, on a bean leaf development was completed in 8.0 days. The longest developmental times on Multicel were recorded on Ephestia eggs (7.0 days) and on decapsulated Artemia cysts (7.5 days). No development beyond the protonymphal stage occurred in the absence of food or when encapsulated Artemia cysts or thrips larvae were offered on Multicel. On a sweet pepper leaf and a bean leaf, respectively 6.7 and 10.0% of the eggs reached adulthood when thrips larvae were provided as food; developmental times recorded here, were 9.0 and 8.3 days. Overall, immature mortality occurred mainly in the protonymphal stage and ranged from 0.0 to 36.7%. In conclusion, I. degenerans is able to feed on a variety of natural and unnatural foods, but thrips larvae and sweet pepper pollen are unfavourable food for immature development. This could compromise the establishment of this biological control agent when used against thrips in sweet pepper crops.     

Confirmation and identification of parasitic stages of obligate endobionts (Harpellales) in blackflies (Simuliidae) by means of rRNA sequence data

Over the last 35 y, the life cycle of endosymbiotic gut fungi (Harpellales) has been expanded to include cyst stages associated with the developing ovaries. Ovarian cysts (chlamydospore stages) have been identified after germination and production of asexual trichospores in vitro, but germination is not always successful, and spores exhibit morphological variation. Sequence data (for partial 18S and 28S rRNA genes) were generated for these putative stages of Harpellales using ungerminated cysts from adult blackflies and germinated cysts associated with field-collected blackfly egg masses. Cladistic analyses of the 18S and 28S rRNA sequences confirmed that ovarian cysts are stages in the life cycle of Harpellales. Ungerminated cysts, from a blackfly collected from New York state were identified as Pennella simulii and two samples from Newfoundland Prosimulium mixtum adults were identified as Harpella melusinae. Cysts with bipolar germ tubes, associated with field-collected Simulium egg masses from Newfoundland, were also identified as H. melusinae. Two other samples of cysts could not be matched with available sequences of gut fungi from larval hosts. The potential use of this approach to identify pathogenic stages associated with adult ovaries or field-collected egg masses among other host groups is highlighted and promoted as a tool to test the hypothesis that ovarian cysts are a dispersal stage common to all genera of Harpellales.     

Susceptibility of Meligethes spp. and Dasyneura brassicae to entomopathogenic nematodes during pupation in soil

The susceptibility of pupating larvae of pollen beetles, Meligethes spp. Stephens (Coleoptera: Nitidulidae) and brassica pod midges, Dasyneura brassicae Winnertz (Diptera: Cecidomyidae) to entomopathogenic nematodes (Nematoda: Rhabditida) was studied in the laboratory. The results showed that brassica pod midge larvae were almost unaffected by the tested nematodes (Steinernema bicornutum, S. feltiae and Heterorhabditis bacteriophora) whereas successful pupation of pollen beetle larvae was reduced with increasing number of nematodes (S. bicornutum, S. carpocapsae, S. feltiae and H. bacteriophora). The exposed larvae had been collected in the field and some of the pollen beetle larvae were parasitised by parasitoid wasps. It appeared that parasitised larvae were less affected by nematodes than non-parasitised larvae.     

The foraging behaviors of larvae of the ladybird beetle, <Emphasis Type="Italic">Coccinella septempunctata</Emphasis> L., (Coleoptera: Coccinellidae) towards ant-tended and non-ant-tended aphids

The foraging behaviors of larvae of the ladybird, Coccinella septempunctata L., towards both the ant-tended aphid, Aphis craccivora Koch, and the non-ant-tended aphid, Acyrthosiphon pisum Harris, were investigated in the field and in laboratory experiments. Although there were no differences in the development and growth of the ladybird larvae that preyed on either Ac. pisum or Ap. craccivora, the foraging efficiency of the ladybird larvae that preyed on Ap. craccivora was higher than that of the ladybird larvae that preyed on Ac. pisum in the absence of ants. This result was explained by the fact that the number of Ac. pisum that escaped by dropping off the plant was conspicuously larger than the number of Ap. craccivora that escaped in this fashion and derived from the non-ant-attendance associated with Ac. pisum. In the laboratory experiments, fewer ladybird larvae climbed onto a plant with Ap. craccivora in the presence of ants than onto a plant with Ac. pisum in the absence of ants. The ladybird larvae did not switch from foraging for Ap. craccivora to foraging for Ac. pisum, even after suffering attacks by ants on a plant with Ap. craccivora, and it would appear that ladybird larvae are unable to remember where they have previously been attacked by ants. These results could explain why the ladybird larvae in the field more frequently visited Vicia angustifolia plants with Ap. craccivora than those with Ac. pisum and made more visits when ants were absent than when they were present.     

Parasitoid Preference for Host-Infested Plants Is Affected by the Risk of Intraguild Predation

Flowering Rorippa indicaplants are attended by ants that collect nectar and, at the same time, prey on herbivorous insects, including larvae of the diamondback moth, Plutella xylostella.Here, we showed that P. xylostellalarvae suffered higher predation on R. indicawhose flowers were accessible by ants than on plants those whose flowers were inaccessible. Ants showed equal predation preference between unparasitized and larvae parasitized by Cotesia plutellae,a dominant specialist parasitic wasp of P. xylostellalarvae. C. plutellaepreferred non-flowering, host-infested R. indicato flowering, host infested R. indica.Based on these results, we infer that the preference of C. plutellaefor non-flowering, host-infested plants is in part explained by the avoidance of intraguild predation by attending ants.     

Identification of a flavobacterium strain virulent against Giardia lamblia cysts

We have isolated from a Kentucky stream a bacterial strain capable of killing the cyst form of Giardia lamblia. This bacterium, designated Sun4, is a Gram-negative, aerobic rod which produces a yellow pigment, but not of the flexirubin-type. Although true gliding motility has not been observed in Sun4, this strain does exhibit a spreading colony morphology when grown on R2A agar. Strain Sun4 has been identified by 16S rRNA sequencing and phylogenetic analysis as belonging to the genus Flavobacterium, and is most closely related to Cytophaga sp. strain Type 0092 and associated Flavobacterium columnare strains. Lipid analysis also identified fatty acids characteristic of the Cytophaga–Flavobacterium group of bacteria. In culture, Sun4 is able to degrade casein and cellulose, but not chitin, gelatin, starch, or agar. Degradation of Giardia cysts by Sun4 appears to require direct cellular contact as neither cell-free extracts nor cells separated from the cysts by dialysis membranes showed any activity against cysts. Activity against Giardia cysts is rapid, with Sun4 killing over 90% of cysts within 48 h. Strain Sun4 requires elevated levels of Ca²⁺ for optimal growth and degradative activity against Giardia cysts. We propose that bacterial strains such as Sun4 could be used as biological control agents against Giardia cysts in drinking water treatment systems.     

The brine shrimp Artemia sp. as alternative prey for rearing the predatory bug Macrolophus caliginosus

The predatory bug Macrolophus caliginosus, which is widely used in greenhouse crops, is limited in its application by its high price. An important factor in the cost is the high price of Ephestia kuehniella eggs, the prey used in their mass rearing. In order to reduce their price, alternatives to moth eggs are currently being investigated. The brine shrimp Artemia sp. is produced in large quantities in saline lakes and is fed as live food source to the larvae of a variety of marine and freshwater organisms. In this study, we tested Artemia sp. as prey for rearing M. caliginosus from two strains. We evaluated developmental and reproduction parameters of the predator when fed nauplii, enriched nauplii with a fatty acid, dry cysts and hydrated cysts, and were compared with those obtained when the predator was fed with E. kuehniella eggs. Nauplii had a significant reduction in survivorship, a delay in development of nymphs and a low reproduction of adults. Nauplii enriched with docosahexaenoic acid (DHA, 22:6n − 3), a common practice for larviculture of some marine fish species, resulted toxic to M. caliginosus nymphs and survival was quite low. On the contrary, either dry or hydrated cysts from the two strains tested of the brine shrimp produced the same nymphal survivorship, nymphal development time and weight and fecundity of adults as those obtained with E. kuehniella eggs. Demographic parameters of the eighth generation of the predator reared with cysts of the two strains, either dry or hydrated, were as good as those of moth eggs. We concluded that Artemia sp. cysts were a good substitution prey for the mass rearing of M. caliginosus.     

Response of the wasp (<Emphasis Type="Italic">Cotesia glomerata</Emphasis>) to larvae of the large white butterfly (<Emphasis Type="Italic">Pieris brassicae</Emphasis>)

The large white butterfly (Pieris brassicae L) first invaded northernmost Japan from Siberia around 1994, and after a few years, began to expand its range. The wasp, Cotesia glomerata (L) parasitizes larvae of the small white butterfly (Pieris rapae crucivora Boisduval), a usual host in the same geographic area. Some Pieris brassicae larvae in Hokkaido have been parasitized by Cotesia glomerata, but the parasitism rate of Pieris brassicae larvae tends to be lower than that of Pieris rapae. To examine the process of parasitizing Pieris brassicae larvae, we observed how the parasitoid wasp responded to the host larvae on damaged leaves. Cotesia glomerata females tended to avoid Pieris brassicae larvae, and even when female wasps inserted their ovipositors into Pieris brassicae larvae, none laid eggs. The parasitoids obtained from Pieris rapae larvae failed to parasitize Pieris brassicae during the host-acceptance step.     

Effects of diet on development and reproduction of the predatory pentatomids Picromerus bidens and Podisus maculiventris

The effects of five diets (larvae of Galleria mellonella L., larvae of Spodoptera littoralis (Boisduval), eggs of Ephestia kuehniella Zeller, cysts of Artemia franciscana Kellogg and an artificial diet based on bovine meat) on development, survival and reproduction of two predatory stinkbugs, Picromerus bidens L. and Podisus maculiventris (Say), were studied in the laboratory. Both species successfully completed immature development on the foods offered, except for P. bidens on cysts of A. franciscana. Significant effects of diet on developmental duration were found in both species. Total developmental time from second instar to adult ranged from 25.0 to 41.5 days and from 18.7 to 46.0 days in P. bidens and P. maculiventris, respectively. Nymphal survival of P. maculiventris was superior to that of P. bidens on all diets tested. Nymphal survival of P. maculiventris was greater than 92% on all diets except on A. franciscana cysts, yielding only 50% survival to adulthood. Survival of P. bidens fed on eggs of E. kuehniella was higher than that of conspecifics fed on caterpillar prey or artificial diet (89% on flour moth eggs vs. 68 and 80% on G. mellonella and S. littoralis and only 50% on meat diet). Fresh weight of newly emerged females of both pentatomids was affected by the diet offered to the predators during their nymphal stage. Females of P. maculiventris produced viable eggs on all diets except on cysts of A. franciscana and had mean fecundities of 691, 436, 608 and 344 eggs per female on S. littoralis, G. mellonella, E. kuehniella eggs and the meat diet, respectively. Females of P. bidens laid eggs only on live prey with mean fecundities of 94 and 38 eggs per female on S. littoralis and G. mellonella, respectively. The results indicate a lower nutritional plasticity of P. bidens as compared with P. maculiventris. An erratum to this article can be found at     

Biology, rearing and field release on Guam of Euplectrus maternus, a parasitoid of the fruit-piercing moth, Eudocima fullonia

The eulophid parasitoid, Euplectrus maternus, is a gregarious ectoparasitoid of the larvae of thefruit-piercing moth, Eudocima (Othreis) fullonia. This parasitoid is indigenous to India and was released into Guam in 1998–99 to aid in the biological control of E. fullonia, an important pest of ripeguava, banana, mango, pomegranate, litchi,papaya, tomato, orange and other fruit crops.In the laboratory, biology and rearing of E. maternus on the larvae of E. fulloniawere studied. In no-choice tests, ovipositionby E. maternus was significantly greateron first and second instar larvae than on thirdinstar larvae. However, in free-choice tests,oviposition by E. maternus wassignificantly greater on second instar larvae,followed by first and third instar larvae. Thisparasitoid did not lay eggs on later instars,either in no-choice or free-choice tests.Females laid a significantly greater number ofeggs (>80%) on the dorsal surface of thelarvae than on either lateral or ventralsurfaces (1 to 8%). In general, more eggs werelaid on abdominal segments 1–3 on larvalinstars one to three. The survival of theimmature stages (from egg to adult) of E. maternus on second instar E. fullonialarvae was 61%, which was significantlygreater than the survival rates on first andthird instar larvae (32% and 26%), indicatingthat second instar host larvae are ideal formass rearing of the parasitoid. Mated femaleE. maternus continuously laid eggs on thesecond instar larvae of E. fullonia forup to 30 days, but the greatest number of eggswere laid during the first week after exposure.When E. maternus laid more than two eggson host larva, more female progeny wereproduced, indicating female-biasedreproduction. Adult parasitoids lived longerwhen fed with a honey: water (50% w/v)solution than with pure honey. Similarly, thefecundity of females increased significantlywhen fed with the honey-water solution whencompared to feeding with pure honey. To date,E. maternus has not established in Guam.     

Specificity of an ant-lycaenid interaction

Summary Many lycaenid butterflies are believed to be mutualists of ants — the butterfly larvae secrete sugars and amino acids as rewards for the ants, and the ants protect the larvae from predation or parasitism. We examined the specificity of the relationship between the lycaenid Plebejus argus and ants in the genus Lasius. Eggs were not attractive to Lasius ants until the emerging larvae had broken through the chorion. First instar larvae were palpated and picked up by Lasius workers and taken to the nest. First instars were mostly ignored by Myrmica sabuleti ants and they were rarely detected by Formica fusca. Older larvae were more attractive to Lasius than to the other ant genera. Pupae were very attractive to Lasius, moderately so to Myrmica, and were ignored by Formica fusca. Teneral adults were palpated by Lasius, but were attacked by Myrmica and Formica workers. We conclude that P. argus is a specialist associate of Lasius ants. Two populations of Plebejus argus were compared: one is naturally associated with Lasius niger, and the other with Lasius alienus. In reciprocal trials, larvae were slightly more attractive to their natural host ant species. Since test larvae were reared on a single host plant species in captivity, this differentiation probably has a genetic basis.     

Susceptibility of Delia radicum to steinernematid nematodes

Isolates of different Steinernema species (S. affine, S. bicornutum, S. feltiae and Steinernema C1) were used in mortality assays with third instar larvae of Delia radicum (L.) (Diptera: Anthomyiidae). The nematode isolates had been obtained by baiting soil regularly grown with cabbage. One isolate (S. feltiae) was the result of a natural infection of a D. radicum puparium. The highest mortality (77%) was obtained with an isolate of S. feltiae (DK1). The isolate DK1 was also used in tests with all larval stages of D. radicum. Mortality around 60% was observed for second and third instar larvae, while first instar larvae showed very low or no susceptibility. Maximum mortality of second and third instar larvae was reached applying only 25 nematodes per larva. Observations of larvae that pupated revealed that some of these puparia contained nematodes. Experiments with hatching puparia showed that a high proportion was infected by nematodes if the flies were prevented from leaving nematode-containing soil. In addition to mortality, the ability of the nematodes to successfully reproduce in the insects was studied. It was found that the species S. feltiae and S. bicornutum reproduced in D. radicum larvae and adults with S. feltiae being the most successful.     

Dinoflagellate cyst production in one-liter containers

Methods for the production of dinoflagellate cysts in two types of 1 L containers have been developed. Using these methods, dinoflagellate cysts can be produced in amounts large enough for shellfish grazing experiments or whenever large amounts of cysts are needed. The species used were Scrippsiella lachrymosa (B-10) and toxic Alexandrium fundyense (CB501 and GTM25). Cultures of S. lachrymosa yielded 628 ± 74 cysts mL^–1 and A. fundyense cultures yielded 350 ± 98 cysts mL^–1. Findings suggest that aspects of the boundary layer between the media and the wall of the container are important for gamete mating; especially, the slope of the container wall appears to be relevant, which offers some explanation of previous observations that the shape of the container is important in the formation of dinoflagellate resting cysts. These observations may support the theory that physical interfaces in nature facilitate dinoflagellate encystment.     

Behavioural response of Heterorhabditis megidis towards plant roots and insect larvae

The behavioural response of infective juveniles (IJs) of Heterorhabditis megidis (strain NLH-E87.3) to cues from roots of strawberry (Fragaria x ananassa Duch.), thuja (Thuja occidentalis L.) and to larvae of the black vine weevil, Otiorhynchus sulcatus, was studied. Choice assays were conducted in an Y-tube olfactometer filled with moist sand. Infective juveniles were activated by the presence of intact roots of both strawberry and thuja plants. Some nematodes aggregated in the compartments with roots but most moved away from the roots to the opposite side. Given a choice, IJs showed a preference for strawberry roots above O. sulcatus larvae. No difference in preference was observed between thuja roots and O. sulcatus larvae. The combination of strawberry roots with vine weevil larvae was preferred above roots alone. In the assays with thuja roots and larvae versus thuja roots alone, however, IJs were stimulated to move but showed preference for the opposite compartment away from the arms with roots and larvae. Nematodes responded differently to mechanically damaged roots as opposed to roots damaged by vine weevil larvae. In assays with damaged thuja roots, IJs were most attracted by the roots damaged by larvae, whereas in the strawberry assays IJs showed a clear preference for the mechanically damaged roots. When challenged with a choice between strawberry and thuja roots, IJs moved preferentially to strawberry than to thuja roots. A preference for the combination of strawberry roots plus larvae over the thuja roots plus larvae was also observed.     

Impact of flowering buckwheat on Lepidopteran cabbage pests and their parasitoids at two spatial scales

We assessed the potential of annual buckwheat, Fagopyrum esculentum Moench, to lead to improved parasitism of lepidopteran cabbage pests over four years. Pest, parasitism, and hyperparasitism rates were monitored in replicated cabbage plots (12 × 20 m) with or without 3 m wide buckwheat borders from 2000 to 2003. Floral borders did not significantly increase egg, larval, or pupal densities of cabbage looper, Trichoplusia ni (Hübner), imported cabbageworm, Pieris rapae (L.), or diamondback moth, Plutella xylostella (L.). Buckwheat increased parasitism rates by Voria ruralis (Fallen) on T. ni larvae and Cotesia rubecula (Marshall) on P. rapaelarvae over four years. Parasitism by Diadegma insulare (Cresson) on P. xylostella larvae was higher in buckwheat than control plots in the first year, and parasitism by Euplectrus plathypenae (Howard) on T. ni larvae was lower in buckwheat than control plots in the second year. The hyperparasitoid Conura side (Walker) attacked D. insulare all four years, but buckwheat did not affect hyperparasitism rates. The effect of spatial scale on pest densities and parasitism in 2001 was evaluated by comparing plots separated at least 67 m (nearby) versus 800 m apart (isolated). T. ni pupae and P. rapae eggs and pupae were more abundant in plots in closer proximity, whereas P. xylostella densities did not vary by the spatial separation of plots. Tachinids and Pteromalus puparum (L.) attacked more P. rapae in nearby plots. E. plathypenae responded to the treatment × scale interaction, parasitizing more in control than buckwheat when plots were isolated but not when plots were nearby.     

Comparison of gregariousness in larvae and adults of four species of zetoborine cockroaches

Insects observed in groups in nature may be gathered either by attractants from the environment or by conspecifics. Field distribution data alone are thus insufficient to assess congregation by conspecifics and complementary laboratory tests of spacing patterns are required. Such tests were performed in four species of Zetoborinae (Insecta, Blattaria), for which field studies showed differences in spatial distribution (Schultesia lampyridiformis Roth, Phortioeca nimbata Burmeister, Lanxoblatta emarginata Burmeister and Thanatophyllum akinetum Grandcolas). Gregariousness, mobility, and sticking to shelters were compared between these four species. Tests were performed on pairs of adults of the same sex and opposite sexes, and also on first instar larvae, either in isolation or in groups of two and four individuals. In S. lampyridiformis, adults were strongly gregarious, whereas larvae dispersed early after birth and were very mobile during the first instar. In P. nimbata, larvae and adults were gregarious, while in L. emarginata larvae were weakly gregarious, and gregariousness decreased when density increased. Adults of L. emarginata seemed to be indifferent to each other. Larvae and adults of T. akinetum were solitary and they actively dispersed. The varying levels of gregariousness among species are discussed according to the known ecological habits in Zetoborinae.     

Application of real-time PCR assay for detection and quantification of Alexandrium tamarense and Alexandrium catenella cysts from marine sediments

The dinoflagellates Alexandrium tamarense (Lebor) Balech and Alexandrium catenella (Whedon and Kofoid) Balech (Dinophyceae) are believed to be the main species responsible for paralytic shellfish poisoning (PSP) all over the world. It is necessary to identify A. tamarense and A. catenella cysts and to monitor their distribution in sediment in order to minimize the damages caused by PSP to the economy and food quality because cysts are the seed population for blooms caused by motile vegetative cells. In this study, we developed an efficient DNA extraction method from the natural cysts present in marine sediments after they were size fractionated with a plankton net (mesh size of 20–150 μm). The 10–3000 cysts were added to the sediments collected from the Ariake Sea, and for which the primuline-staining method did not reveal any cysts. DNA was then extracted from each sample, and linear standard curves for A. tamarense and A. catenella cysts were obtained from the correlation between the Ct values by real-time PCR and the log of the initial densities of cysts. We monitored the A. tamarense and A. catenella cyst densities in the environmental samples. This assay was demonstrated to be a powerful tool for the identification, detection, and quantification of the cysts of the toxic dinoflagellates.     

Removal of wastewater pathogen indicators in a constructed wetland in Leon, Spain

This paper studies the seasonal presence and removal of the pathogenous micro-organisms Escherichia coli, total coliforms (TC), Clostridium perfringens (Cp), faecal streptococci (FS), Giardia cysts, Cryptosporidium oocysts and helminth eggs, in a constructed wetland treatment system. The removal efficiency of this system with respect to the indicator micro-organisms achieved maximum values in spring and autumn at 99.9% for E. coli and TC, respectively, in winter at 97.0% for FS, in summer at 100% for Clostridium and throughout the year, also at 100%, in the case of Giardia cysts, Cryptosporidium oocysts and helminth eggs. In general, very low protozoan and helminth egg counts were found, and the system demonstrated efficient reduction of the wastewater indicator pathogens.