Tolypocladium cylindrosporum (Deuteromycotina: Moniliales), a fungal pathogen of the mosquito Aedes australis

A blastoconidial preparation of the New Zealand isolate of T. cylindrosporum was tested in two field trials against larvae of the freshwater mosquito Aedes subalbirostris and the brackish water species Ae. australis. There was some reduction in the population of the Ae. subalbirostris larvae and larvae collected from the test pools exhibited signs of fungal infection when reared in the laboratory. In the brackish pools there was no observable decrease in the larval populations even though Ae. australis is the natural host of T. cylindrosporum [NZ]. Water volume fluctuations in the test pools were a major difficulty experienced when monitoring the results.Tests against field-collected non-target fauna indicated that T. cylindrosporum is non-specific in action. Species of daphnid, dixid larvae and copepods were all susceptible to infection at varying degrees.     

Coelomomyces opifexi Pillai & smith (Coelomomycetaceae: Blastocladiales) V. Development of the fungus in pupae and adults of Aedes australis

Abstract

Late-instar Aedes australis larvae were experimentally infected with Coelomomyces opifexi, and the subsequent development of the fungal parasite was studied. Five separate experiments involving 1190 infected larvae were conducted. Of these larvae, 114 continued development and pupated; 38 of them contained sporangia. Histological studies indicated that parasite development slowed down, if not ceased, following pupation of infected larvae, and the presence of sporangia in adults reflected the degree of advancement of infection in the larvae at pupation.     

Host range and specificity of an Argentinean isolate of the aquatic fungus <Emphasis Type="Italic">Leptolegnia chapmanii</Emphasis> (Oomycetes: Saprolegniales), a pathogen of mosquito larvae (Diptera: Culicidae)

An isolate from Argentina of the fungal mosquito pathogen Leptolegnia chapmanii (ARSEF 5499), was tested against 12 species of mosquito larvae and on species of non-target aquatic invertebrates and vertebrates. The mosquito species tested were Aedes aegypti, Anopheles sp., Culex apicinus, Cx. castroi, Cx. dolosus, Cx. pipiens, Cx. renatoi, Isostomyia paranensis, Ochlerotatus albifasciatus, Oc. crinifer, Psorophora cyanescens, and P. ferox. Mosquito larvae of 10 species were susceptible, with mortality rates from 10–100%. Two mosquito species Cx. renatoi and I. paranensis were not infected by Leptolegnia. None of the non-target fauna treated was infected by L. chapmanii with exception of members of the Family Chironomidae which were susceptible at low infection rates. Researcher CIC (Committee for Scientific Research of Buenos Aires State).     

Pathogenicity of Heterorhabditis nematodes isolated from north-western Himalaya against the larvae of Plutella xylostella (L.) (Lepidoptera: Plutellidae)

The efficacy of three entomopathogenic nematodes (Heterorhabditis spp.), from north western Himalaya, India was studied against the diamondback moth, Plutella xylostella (Linnaeus, 1758) (Lepidoptera: Plutellidae), under laboratory conditions. The larvae were exposed to 10, 20, 30 and 40 infective juveniles (IJs) of each nematode species for different time periods and they were found to be susceptible to all the EPNs tested. However, the susceptibility of larvae to nematode infection varied according to the dosages of IJs and their exposure periods. The efficacy of these indigenous entomopathogenic nematodes was also evaluated against the commercially available entomopathogenic nematode H. indica. An indigenous isolate, H. bacteriophora (HRJ), along with the commercial isolate H. indica recorded 100.0% mortality of insect larvae in 96 h exposure time against third instar larvae of P. xylostella. However, it was noticed that with the advancement of larval stage its mortality rate reduces and vice versa with the exposure period. All the tested nematode species were also found to reproduce within the host and produced infective juveniles. In conclusion, the evidence obtained in this study suggests that all the three indigenous EPN species are virulent enough to produce 100% mortality of larvae of P. xylostella. These EPN species thus have potential for the management of P. xylostella under integrated management practices.     

Mouthparts of larvae of Opifex fuscus and Aedes australis (Diptera: Gulicidae); a scanning electron microscope study

Abstract

Larvae of the New Zealand culicid species Opifex fuscus and Aedes australis have previously been reported to show dimorphism in the structure of their labral brushes, some larvae having pectinate bristles and others only simple hairs. In the scanning electron microscope all larvae showed some degree of pectination of hairs in these brushes. There is also a gradation in the pectination. Some bristles are only sparsely pectinate; because the dimensions of their teeth are close to the limit of resolution by the compound microscope, the pectination had previously gone undetected. The mouthparts of both species are intermediate in character between those typical of filter-feeding larvae and those typical of browsing larvae. The SEM appearance of maxillary sensoria and bristles on the ventral surfaces of the mandibular brushes is described; the latter bristles comb food particles out of the labral brushes and towards the mouth. Features of the mouthparts are illustrated with scanning electron micrographs.     

Sensitivity of polyphagous (Plodia interpunctella) and stenophagous (Ephestia kuehniella) storage moths to residual insecticides: effect of formulation and larval age

Pyralid moths, Ephestia kuehniella and Plodia interpunctella, are prevalent stored product pests. The insecticides are the main tool to control these moths in the stores. The data describing the response of these moths to insecticides are scarce. The lethal effect of the organophosphate, pyrethroid, and halogenated-pyrrole on moths larvae were compared in laboratory test. The hypothesis was that the very polyphagous P. interpunctella would have generally higher insecticide tolerance than that of the stenophagous E. kuehniella. Different insecticide concentrations were applied onto the inner surface of glass tube vials. Ten larvae of 14 or 21 d old of E. kuehniella and 7 or 14 d old of P. interpunctella were used by treatment. The larval mortality was checked after 24 h of exposure. The mortality was significantly influenced by age of larvae and the groups of chemicals. No differences among the efficacies of the tested formulations with identical active compounds were found, except significant different mortality of E. kuehniella on deltamethrin formulations. A comparison of analytical standards showed that P. interpunctella was less susceptible to the active ingredient pirimiphos-methyl than E. kuehniella, while E. kuehniella was less susceptible to deltamethrin than P. interpunctella. No differences between the two species were observed for chlorfenapyr. We therefore rejected the hypothesis that polyphagy/stenophagy can be a general predictor of insecticide tolerance in the two tested storage moths. The most important finding for effective use was that the young larvae of both species were more susceptible to tested insecticides than older larvae.     

Three microsporidian pathogens infecting Lymantria dispar larvae do not differ in their success in horizontal transmission

We quantified horizontal transmission of three microsporidian pathogens, Endoreticulatus schubergi, Nosema lymantriae and Vairimorpha disparis that infect Lymantria dispar larvae in an experiment using caged, potted oak plants. Despite marked differences in the modes of spore release from infectious hosts, no significant differences in the transmission success to uninfected, susceptible test hosts were ascertained between the tested microsporidian species. The density of initially inoculated larvae and the exposure period, on the other hand, did influence the number of infected test larvae. Depending on the density of inoculated larvae (10%, 30% or 50%), between 0% and 26% of the test larvae became infected with one of the three tested microsporidian pathogens after an exposure period of 6 days. When the exposure period was 12 days, between 11% and 76% of the test larvae became infected.     

Blocking Phragmites australis reinvasion of restored marshes using plants selected from wild populations and tissue culture

This study tested a vegetation strategy for controlling Phragmites australis invasion into brackish marshes as an alternative to the current technique of repeated herbicide sprays followed by burning. This strategy involves blocking P. australis by planting desired plants selected from wild populations and/or tissue culture regenerants at key points on the major routes of P. australis invasion. The planting of native species was conducted at three sites in a herbicide-treated P. australis marsh near Salem, NJ. Wild population selections of three upland marsh shrubs, Myrica cerifera, Baccharis halimifolia, and Iva frutescens, as well as two grass species, Spartina alterniflora and Spartina patens, and two rushes, Juncus gerardi and Juncus roemerianus, were planted according to their normal zonation positions. Tissue culture regenerated plants of the two grasses and two rushes, and the sedge species Scirpus robustus, were also planted. Plant growth at each site was monitored each year after planting for up to 3 years. Most plants of B. halimifolia, I. frutescens, J. roemerianus, and S. patens demonstrated a consistent vigorous growth at all three sites, whether or not the plants were collected from wild populations or were tissue culture regenerants. These multi-layered walls of plants demonstrated effectiveness in controlling the P. australis by restricting or inhibiting its spread. Upon screening 48 regenerated plants of S. patens at one of the three sites, we found that some regenerants showed enhanced characteristics for blocking P. australis, such as greater expansion and a high stem density. The availability of the tissue culture-regenerated plants of the native marsh species makes it possible to select lines from local genotypes that have desirable characteristics for wetland restoration projects, such as blocking P. australis reinvasion.     

The infectivity of a nuclear polyhedrosis virus of the velvetbean caterpillar for eight noctuid hosts

Eight species of noctuid larvae were tested for susceptibility to a nuclear polyhedrosis virus of the velvetbean caterpillar, Anticarsia gemmatalis. Velvetbean caterpillar larvae were highly susceptible to crude preparations of polyhedral inclusion bodies (PIBs; LD₅₀ = 4.7 PIBs/larva), but preparations of purified polyhedra were much less effective against these larvae (LD₅₀ = 319.7 PIBs/larva). Of seven other noctuid species tested, only Heliothis virescens was as susceptible to the virus as A. gemmatalis. High dosages were required to kill Heliothis zea, Trichoplusia ni, Pseudoplusia includens, and Spodoptera ornithogalli. Plathypena scabra and Spodoptera frugiperda were not susceptible.     

Occurrence and Structure of Iron Inclusions in Adipocytes of Larval Lampreys

The occurrence and structure of adipocytes in the larvae of two lamprey species, Geotria australis and Petromyzon marinus, were examined by electron microscopy. Adipocytes from both species possessed large electron-dense inclusions which histochemical and energy dispersal X-ray analyses show as containing iron. The greatest concentration of inclusions in adipocytes was found in the nephric fold of G. australis. While some iron is present in the cytoplasmic matrix as ferritin, the majority is seen in large ammocoetes in membrane-bound dense aggregations of haemosiderin. The wide variety of inclusion types seen in smaller larvae may reflect on the method of formation of these inclusions within the cell. Because of the high level of iron loading in the larval lamprey nephric fold, this readily accessible tissue may provide a valuable model for studies of iron metabolism in vertebrates.     

Laboratory evaluation of Turkish entomopathogenic nematodes for suppression of the chestnut pests,Curculio elephas (Coleoptera: Curculionidae) and Cydia splendana (Lepidoptera: Tortricidae)

The lepidopteran, Cydia splendana, and the coleopteran, Curculio elephas, are the most serious pests of chestnut fruit in Turkey. We evaluated the biological control potential of three Turkish entomopathogenic nematode species, Steinernema feltiae, S. weiseri and Heterorhabditis bacteriophora, against the last instar larvae of C. splendana and C. elephas, both of which occur in the soil from fall (October–November) until mid-summer (August). The optimal temperature for infection, time to death of the hosts, and reproductive potential of the nematodes were determined at 10, 15, 20 and 25°C for both pest species. Cydia splendana was more susceptible to nematode infection than C. elephas. Temperature had a significant effect on the infectivity and development of entomopathogenic nematodes. The cold-adapted S. weiseri and S. feltiae were the most virulent species at 10 and 15°C, whereas the warm-adapted H. bacteriophora was the most effective at 20 and 25°C. In soil pot experiments conducted at 15°C, S. weiseri was the most virulent species against C. elephas and C. splendana. However, our data show that C. elephas larvae had a lower and C. splendana larvae had a higher susceptibility to the nematode species tested. Accordingly, we recommend that future efforts of using entomopathogenic nematodes, especially S. weiseri, be directed against C. splendana and that there be a continued effort to find more virulent nematode isolates against larvae of C. elephas.     

Oviposition site selection by mosquitoes is affected by cues from conspecific larvae and anuran tadpoles

Abstract The oviposition site that a female mosquito selects will influence the fitness of her larvae. We conducted a series of artificial pond experiments to compare the oviposition responses of two species of mosquitoes with the presence of tadpoles, conspecifics and chemical cues from these organisms. The two mosquito species differ markedly in larval ecology. The larvae of one species, Culex quinquefasciatus, co‐occur with numerous freshwater organisms, including tadpoles of Linmodynastes peronii (the striped marsh frog). Larvae of the other mosquito, Ochlerotatus australis, inhabit small brackish rock ponds where the main potential competitors are tadpoles of Crinia signifera (the common eastern froglet). In field trials, females of both mosquito species oviposited significantly more often in water that contained (or had previously contained) conspecific larvae. However, these superficially similar responses were mediated via different pathways: fungicide abolished the response by C. quinquefasciatus but not by O. australis. The two mosquito species also responded differently to cues associated with syntopic tadpoles. The presence of tadpoles did not influence oviposition by C. quinquefasciatus, but O. australis oviposited less often if tadpoles were present. These interspecific differences in oviposition behaviour may be adaptive to differences in larval ecology: competition with tadpoles is likely to be more significant for O. australis than for C. quinquefasciatus. Our findings thus support the hypothesis that mosquitoes oviposit selectively to avoid potential anuran larval competitors.     

Coelomomyces opifexi (Pillai & Smith) Coelomomycetaceae: Blastocladiales. III. The Laboratory Infection of Aedes australis (Erichson) larvae

This paper outlines the techniques used for the laboratory transmission of C. opifexi to its host larva Aedes australis, which is a brackish water mosquito.The main requirements for infection are (a) a suitable medium of low salinity, consisting of natural australis pool water or soil infusion prepared in the laboratory (b) inoculum containing a high concentration of active zoospores and (c) temperatures approaching summer conditions.In australis pool water infected larvae appear from day 9 till day 20 with a steep rise in number at the beginning and another at the end of the cycle. In the soil infusion infection is detectable at the later time of 16 to 28 days with a single peak between days 18 to 20. The early infected larvae are probably eliminated in infusion medium due to the presence of toxic substances.Temperature has a direct effect on the time taken for infection to show up in the larvae. The use of fluorescent light, simulating daylight appears to be uncritical.From the Department of Microbiology, Medical School, University of Otago, Dunedin, New Zealand.From the Department of Microbiology, Medical School, University of Otago, Dunedin, New Zealand.     

The ichthyoplankton of a seasonally closed estuary in temperate Australia. Does an extended period of opening influence species composition?

Fish eggs and larvae were collected monthly between September 1987 and April 1989 from sites throughout the main basin and within the saline regions of the two main tributary rivers of Wilson Inlet, a seasonally closed estuary in south-western Australia. Of the eggs, 43.7% belonged to Engraulis australis (Shaw) and the rest to unidentified teleosts. The larval fish assemblage comprised 17 families represented by 25 species. The Gobiidae contained the highest number of species (four) and contributed approximately 57% of all larvae caught. Pseudogobius olorum (Sauvage) and E. australis were the most abundant species, contributing 43.9 and 27.9% to the total larval catch, respectively. The larvae of species which breed within Wilson Inlet dominated the assemblage, both in terms of number of species (64%) and contribution to total catch (99.9%). The numbers of the eight marine species and one freshwater species represented in the ichthyoplankton were very low. Classification and multi-dimensional scaling ordination showed that the composition of the larval fish fauna at the various sites during a period when the estuary remained open to the sea (December 1988-April 1989) was similar to that of the corresponding sites during the same period in the previous year when the estuary had become closed (December 1987-April 1988). This can be attributed to the spatial distribution, time of occurrence and abundance of estuarine-spawned larvae being similar in the two periods and to the rarity of marine-spawned larvae, even in the spring and summer of 1988/1989 when the estuary was open for the whole time when most marine teleosts spawn in south-western Australia. The low occurrence of marine-spawned larvae in Wilson Inlet reflects the fact that tidal water movement within the basin of the system is so small that it is unable to facilitate the transport and dispersion of larvae. The ichthyoplankton of Wilson Inlet resembles that of other poorly-flushed estuaries in that it is low in species richness and dominated by estuarine-spawned larvae.     

Relative susceptibility of several noctuid species to a nuclear polyhedrosis virus from Heliothis armiger

Several alternate hosts were tested for their relative susceptibility to an isolate of Galleria mellonella nuclear polyhedrosis virus. Neonate Trichoplusia ni, Heliothis zea, and Manduca sexta were all susceptible to per oral administration of purified polyhedra. Of the three alternate species tested, T. ni was the most susceptible, and exhibited the most variable mortality response over the dose range tested, while M. sexta was the least susceptible. We believe this represents the first report of a lethal virus infection in a sphingid species, and useful parameters for the successful inoculation of alternate hosts are discussed.     

Host habitat finding and host selection of theDrosophila parasitoidLeptopilina australis (Hymenoptera,Eucoilidae), with a comparison of the niches of EuropeanLeptopilina species

Summary Decaying petioles of giant hogweed,Heracleum mantegazzianum Sommier & Levier, are used as a breeding site by six species ofDrosophila and the drosophilidScaptomyza pallida. The most numerous parasitoid species associated with this community isLeptopilina australis. BecauseL. australis was previously unknown in western Europe, we present the characters to distinguish it form its close relativeL. clavipes. Experiments on host species selection and survival ofL. australis showed that this parasitoid mainly usesD. limbata as host. Olfactometer experiments showed thatL. australis is attracted by the odour of decaying hogweed stalks, especially when these contain larvae ofD. limbata. L. australis is also strongly attracted by the odour of stinkhorns, a habitat in which it has never been found in nature.D. phalerata is the dominant fly species in stinkhorns, and is not a host ofL. australis. We offer a possible functional explanation for this unexpected habitat choice, by showing thatD. transversa andD. kuntzei, both species found to breed in fungi, are also suitable hosts forL. australis. We also discuss habitat choice with regard to a proposed phylogeny of theLeptopilina species in temperate Europe. Finally, we discuss niche overlap ofL. australis with the otherLeptopilina species.     

Phylogenetic relationships between Lymnaeidae in relation to infection with Fasciola sp. in Hokkaido,Japan

ABSTRACT

In this paper, lymnaeid snails collected from 107 localities in Hokkaido, Japan were examined for infection by Fasciola larvae. DNA sequences from the snails were analysed phylogenetically based on their mitochondrial cytochrome c oxidase (COI) gene. Based on shell morphology, the snails were identified as Galba truncatula, Radix auricularia, and Austropeplea ollula and were divided into distinct clades in the COI tree. No Fasciola larvae were detected in any of the collected snails. The 58 haplotypes of Galba truncatula which were found in Hokkaido included five detected previously in snails infected with Fasciola larvae, and constituted a closely related haplogroup (Gt-c1), suggesting that Gt-c1 snails would be susceptible to Fasciola sp. The Gt14 haplotype snails were considered to be Lymnaea schirazensis, an intermediate host of F. hepatica. The COI haplotypes of A. ollula belonged to a single clade, and were considered conspecific, whereas those of R. auricularia constituted three genetically distinct haplogroups, each of which was possibly a distinct species. This study suggests that morphologically-identified lymnaeid snails possibly include additional species. Further study of these snails is required to clarify their susceptibility to Fasciola sp.     

The fauna of the South China Sea include unknown phoronid species: new records of larvae and adults

The Phoronida is a phylum of marine invertebrates that have a worldwide distribution and that form huge benthic aggregations in many aquatic areas. Although there are only 11 recognized species of phoronids, many species clearly remain to be described. The matching of larval and adult stages of the same species will help indicate which larvae belong to described species and which represent undescribed species whose adults have yet to be found. Larvae of four phoronid species were collected in Nha Trang Bay and investigated by light microscopy and molecular methods. Three of the described larvae belong to new phoronid species that have yet to be named. These larvae have unique morphological features and occupy separate positions on the phylogenetic tree of phoronids. One of the described larvae belongs to Phoronis hippocrepia or to a closely related species. Observations made with a new macrophotographic method indicated that Phoronopsis californica also occurs in Nha Trang Bay. Thus, according to records of larvae and adults, Nha Trang Bay contains at least nine phoronid species: Phoronopsis californica, Phoronopsis malakhovi, Phoronis australis, Phoronis hippocrepia (or a closely related species), three unknown phoronid species whose larvae have unique morphological features and that apparently belong to the genus Phoronis, and two species, whose adults have unique set of morphological features. The richness of phoronids in this area suggests that the Indo-Pacific is a centre of phoronid biodiversity.     

Comparative laboratory studies on three fungal pathogens of the elm bark beetle Scolytus scolytus: Pathogenicity of Beauveria bassiana,metarhizium anisopliae, and Paecilomyces farinosus to larvae and adults of S. scolytus

The entomogenous fungi Beauveria bassiana (nine isolates), Metarhizium anisopliae (seven isolates), and Paecilomyces farinosus (four isolates) were tested as pathogens of larvae of the elm bark beetle, Scolytus scolytus. Single isolates of B. bassiana and M. anisopliae were also tested against adult beetles. Of the 21 isolates tested as conidial suspensions against larvae, all proved pathogenic. The three most and least virulent isolates were, respectively, isolates of B. bassiana and M. anisopliae. The other isolates fell between these two extremes, with the four P. farinosus isolates all moderately virulent. Spore retention on larvae following inoculation was estimated by washing conidia off the larvae. From the results it was possible to relate larval mortality to the approximate spore dose causing infection at different spore concentrations. Thus, application of spores of the three pathogens at a concentration of 10³ spores/ml resulted in limited mortality. At this concentration, an average of only a single spore was recovered from the inoculated larva. Adult bark beetles also proved susceptible to infection by isolates of B. bassiana and M. anisopliae. They were exposed to discs of elm bark dipped in a conidial suspension. It was estimated that a dose of less than 100 spores could cause infection of beetles following feeding on the elm bark discs.