Persistence and expression of Cotesia congregata polydnavirus in host larvae of the tobacco hornworm, Manduca sexta

The gregarious braconid wasp Cotesia congregata parasitizes host larvae of Manduca sexta, and several other sphingid species. Parasitism induces host immunosuppression due to the disruptive action of the wasp's polydnavirus (PDV) on host blood cells. During the initial stages of parasitism, these cells undergo apoptosis followed by cell clumping, which clears the hemolymph of a large number of cells. In this study, the persistence and expression of Cotesia congregata PDV (CcPDV) were examined using Southern and Nor-them blots, respectively. Digoxygenin-labelled total polydnaviral DNA was used to probe genomic DNA isolated from fat body and brains of hosts with emerged wasps taken 6 days following egress of the parasitoids, and significant cross-hybridization between the host fat body genomic DNA with viral DNA was seen. Thus, the virus persists in the host for the duration of parasitism. even during the post-emergence period, and may even be integrated in the host caterpillar DNA. Viral gene expression was examined using Northern blots and probes to the Cotesia rubecula CrV1 homolog, and the CrV1-like mRNAs were expressed as early as 4 h post-parasitization for at least 72 h and faint hybrization is even seen at the time the wasps eclose. In contrast, in Pieris rapae larvae the CrV1 transcript is expressed only for a brief time, during which time hemocyte function is disrupted. The effect is transitory, and hemocytes regain their normal functions after the parasites emerge as first instars. The genome of CcPDV contains one copy of the CrV1-like homolog as shown on Southern blots of viral genomic DNA. In conjunction with our earlier studies of the PDV-encoded early protein 1, the current work suggests multiple viral transcripts are produced following parasitization of the host. and likely target host hemocytes to induce their apoptosis, thereby preventing encapsulation of the parasitoid's eggs. Whether viral DNAs are integrated in the host's genomic DNA remains to be proven, but our results provide preliminary evidence that viral DNAs are detected in the host's fat body cells examined at the time of wasp ernergence and several days later. (C) 2003 Elsevier Science Ltd. All rights reserved.

Variation in the dispersal potential of non-feeding invertebrate larvae: the desperate larva hypothesis and larval size

For many species of marine invertebrates, variability in larval settlement behaviour appears to be the rule rather than the exception. This variability has the potential to affect larval dispersal, because settlement behaviour will influence the length of time larvae are in the plankton. Despite the ubiquity and importance of this variability, relatively few sources of variation in larval settlement behaviour have been identified. One important factor that can affect larval settlement behaviour is the nutritional state of larvae. Non-feeding larvae often become less discriminating in their 'choice' of settlement substrate, i.e. more desperate to settle, when energetic reserves run low. We tested whether variation in larval size (and presumably in nutritional reserves) also affects the settlement behaviour of 3 species of colonial marine invertebrate larvae, the bryozoans Bugula neritina and Watersipora subtorquata and the ascidian Diplosoma listerianum. For all 3 species, larger larvae delayed settlement for longer in the absence of settlement cues, and settlement of Bugula neritina larvae was accelerated by the presence of settlement cues, independently of larval size. In the field, larger W subtorquata larvae also took longer to settle than smaller larvae and were more discriminating towards settlement surfaces. These differences in settlement time are likely to result in differences in the distance that larvae disperse in the field. We suggest that species that produce non-feeding larvae can affect the dispersal potential of their offspring by manipulating larval size and thus larval desperation.

Description of the larvae of Synoestropsis furcata Flint, 1974 (Trichoptera: Hydropsychidae) from central-western region, Brazil

The genus Synoestropsis contains 10 described species, six of which occur in Brazil. The species S. furcata was described by Flint (1974), based on adults only. In this paper, the larvae are described and illustrated. This description constitutes the first definitive species-level association in the genus, and also enlarges the known distribution for S. furcata.

Description of the immature stages of Notalina morsei Holzenthal, 1986 (Trichoptera : Leptoceridae) and an updated key to larvae of Neotropical Leptoceridae genera

The endemic Neotropical long-horned caddisfly subgenus Notalina (Neonotalina) Holzenthal contains nine described species, but its immature stages are unknown. In this paper the larvae and pupae of Notalina morsei Holzenthal 1986 from southeastern Brazil are described and illustrated. Larvae of the subgenus are easily recognized from other Neotropical leptocerids by the following characters: ventral apotome which is broad anteriorly and narrow posteriorly; the metanotum with three sclerites; the metasternum bearing 10-12 setae; the gill arrangement, usually including ventral and dorsal filaments from abdominal segments II to VI; and abdominal tergite IX with 6 long and 4 short setae. An updated key to known larvae of Neotropical Leptoceridae genera is provided.

Excretion of ammonia by Lucilia cuprina larvae suppresses immunity in sheep

Attempts to immunise sheep against natural infestations by Lucilia cuprina larvae have not been effective. Yet it is known that the larvae excrete the immunosuppressant ammonium bicarbonate. The effect of larval ammonium and nonionic ammonia on immunopathobiology was evaluated in 12 infested sheep. The concentration of ammonium in veins draining infested sites was measured in another group of four sheep. Mean jugular unionized ammonia concentration increased 3.5 to 5.6 times above pre-infested control levels. Mean venous ammonium concentrations draining infested sites were 13 times higher than pre-infested jugular or carotid levels. Increases in jugular ammonia concentrations correlated with increased number of larvae, area of infestation, earlier death, neutropenia, eosinopenia, lymphocytopenia, large declines in serum globulins and zinc, and large rises in toxic neutrophils. The high concentrations of toxic unionized ammonia in blood directly permanently damaged neutrophils and lymphocytes and depressed serum globulin production. The results show that the ammonium from the excreta of larvae of L. cuprina may be highly immunosupressive. (C) 1997 Elsevier Science B.V.

Novel gene containing multiple epidermal growth factor-like motifs transiently expressed in the papillae of the ascidian tadpole larvae

We have investigated molecular mechanisms of the embryonic development of an ascidian, a primitive chordate which shares features of both invertebrates and vertebrates, with a view to identifying genes involved in development and metamorphosis, We isolated 12 partial cDNA sequences which were expressed in a stage-specific manner using differential display, We report here the isolation of a full-length cDNA sequence for one of these genes which was specifically expressed during the tailbud and larval stages of ascidian development, This cDNA, 1213 bp in length, is predicted to encode a protein of 337 amino acids containing four epidermal growth factor (EGF)-like repeats and three novel cysteine-rich repeats, Characterization of its spatial expression pattern by in situ hybridisation in late tailbud and larval embryos demonstrated strong expression localised throughout the papillae and anteriormost trunk and weaker expression in the epidermis of the remainder of the embryo, As recent evidence indicates that the signal for metamorphosis originates in the anterior trunk region, these results suggest that this gene may have a role in signalling the initiation of metamorphosis. (C) 1997 Wiley-Liss, Inc.

The insulin signaling pathway in honey bee (Apis mellifera) caste development - differential expression of insulin-like peptides and insulin receptors in queen and worker larvae

The insulin/insulin-like signaling (IIS) pathway is an evolutionarily conserved module in the control of body size and correlated organ growth in metazoans. In the highly eusocial bees, the caste phenotypes differ not only in size and several structural features but also in individual fitness and life history. We investigated the developmental expression profiles of genes encoding the two insulin-like peptides (AmILP-1 and AmILP-2) and the two insulin receptors (AmInR-1 and AmInR-2) predicted in the honey bee genome. Quantitative PCR analysis for queen and worker larvae in critical stages of caste development showed that AmILP-2 is the predominantly transcribed ILP in both castes, with higher expression in workers than in queens. Expression of both InR genes sharply declined in fourth instar queen larvae, but showed little modulation in workers. On first sight, these findings are non-intuitive, considering the higher growth rates of queens, but they can be interpreted as possibly antagonistic crosstalk between the IIS module and juvenile hormone. Analyzing AmInR-1 and AmInR-2 expression in ovaries of queen and worker larvae revealed low transcript levels in queens and a sharp drop in AmInR-2 expression in fifth instar worker larvae, indicating relative independence in tissue-specific versus overall IIS pathway activity. (C) 2008 Elsevier Ltd. All rights reserved.

In vitro study of the antimicrobial activity of Brazilian propolis against Paenibacillus larvae

The honey bee disease American foulbrood (AFB) is a serious problem since its causative agent (Paenibacillus larvae) has become increasingly resistant to conventional antibiotics. The objective of this study was to investigate the in vitro activity of propolis collected from various states of Brazil against P. larvae. Propolis is derived from plant resins collected by honey bees (Apis mellifera) and is globally known for its antimicrobial properties and particularly valued in tropical regions. Tests on the activity of propolis against P. larvae were conducted both in Brazil and Minnesota, USA using two resistance assay methods that measured zones of growth inhibition due to treatment exposure. The propolis extracts from the various states of Brazil showed significant inhibition of P. larvae. Clear dose responses were found for individual propolis extracts, particularly between the concentrations of 1.7 and 0.12 mg propolis/treatment disk, but the source of the propolis, rather than the concentration, may be more influential in determining overall activity. Two of the three tested antibiotics (tylosin and terramycin) exhibited a greater level of inhibition compared to most of the Brazilian samples, which could be due to the low concentrations of active compounds present in the propolis extracts. Additionally, the majority of the Brazilian propolis samples were more effective than the few collected in MN, USA. Due to the evolution of resistance of P. larvae to conventional antibiotic treatments, this research is an important first step in identifying possible new active compounds to treat AFB in honey bee colonies. (C) 2007 Elsevier Inc. All rights reserved.

Ecological distribution of mosquito larvae of the Anopheles punctulatus group on Niolam (Lihir) Island, Papua New Guinea

We surveyed the larval habitats of member,, of the Anopheles punctulatus group of mosquitoes on Niolam (Lihir) Island. Papua New Guinea. Identification of this group was undertaken by polymerase chain reaction-restriction fragment length polymorphism analysis of the amplified internal transcribed spacer unit 2 of rDNA. because morphologic separation of member species is unreliable. The most widespread malaria vector species and their most common larval habitats identified to aid source-reduction programs for malaria control. The most ubiquitous species was An. punctulatus, followed by An. farauti no. 2. then An. farauti s.s. Anopheles punctulatus has increased relative to An.farauti s.l. since the start of development projects on Lihir Island. The most common larval habitats were shallow temporary pools with clay substrate and with plants or floatage. These habitats. mostly encountered alongside poorly drained roads, may be increased by development projects.

Isolation and identification of the toxic peptides from Lophyrotoma zonalis (Pergidae) sawfly larvae

The broad-leaved paper bark tree Melaleuca quinquenervia (Cav) (Myrtaceae) was introduced into Florida (USA) early in this century it has proliferated to such an extent that urgent measures are now required to control it. The sawfly Lophyrotoma zonalis (Pergidae) has been introduced as a possible biological control agent due to its ability to defoliate M. quinquenervia. Because toxic D-amino acid- containing peptides have been isolated from some sawfly species, L. zonalis larvae were processed using the previously reported method for the recovery of these compounds. The toxins lophyrotomin (as the free C-terminal acid) and a mixture of pergidin and Val(4)-pergidin were isolated at 0.36 and 0.43% yield of the dried larvae, respectively. Both compounds when dosed intraperitoneally to C57/B16 male mice were hepatotoxic with lowest lethal doses of 8 and 32 mg/kg, respectively. The pathology of the liver was different for each compound, with the lophyrotomin free acid causing a periportal haemorrhagic necrosis and the pergidin causing a periacinar coagulative necrosis. (C) 2001 Elsevier Science Ltd. All rights reserved.

Detrimental effects of latex and cardiac glycosides on survival and growth of first-instar monarch butterfly larvae Danaus plexippus feeding on the sandhill milkweed Asclepias humistrata

1. A novel experimental method was developed to study negative physical and chemical effects of latex and cardiac glycosides on first-instar monarch butterfly larvae in their natural environment in north central Florida. Forceps were used to nibble through the petioles of leaves of the sandhill milkweed Asclepias humistrata, mimicking the behaviour of mature monarch larvae. This notching cut off the supply-of latex to the leaves without significantly reducing either their cardiac glycoside concentration or water content. 2. The mean cardiac glycoside concentration in larvae that fed on intact leaves was nearly two: and a half times greater than in larvae that fed on notched leaves. This was probably because more latex is present in the gut of the larvae that fed on the intact leaves. Supporting this is the fact that the mean concentration of cardiac glycosides in the latex was 34-47 times that in the leaves. 3. Wet weights, dry weights, and growth rates of first-instar larvae that fed on intact leaves over a 72-h period were less than half those of larvae that fed on notched leaves. 4. Mortality due to miring in the latex was 27% on the intact leaves compared with 2% on the notched leaves. 5. Latex, cardiac glycosides, and other as yet undetermined plant factors all have a negative effect on:first-instar larval survival. 6. Video-analyses indicated that ingestion of latex caused the larvae to become cataleptic and increased their chances of being mired on the leaf by the setting latex glue. Dysfunction resulting from latex ingestion may lead to the larvae falling off the plant and being killed by invertebrate predators. 7. The difficulty of neonate monarch larvae surviving on A. humistrata - one of the principal milkweed species fed on each spring as monarchs remigrate from Mexico into the southern U.S.A. - is evidence that a co-evolutionary arms race is operating in this plant-herbivore system.

Winter intervention against Aedes aegypti (Diptera : Culicidae) larvae in subterranean habitats slows surface recolonization in summer

At semiarid Charters Towers, north Queensland, Australia, the importance of Aedes aegypti (L.) in wells was assessed in relation to the colonization of surface habitats during the wet season. From April to July 1999, 10 wells (five positive for Ae. aegypti) were monitored to assess their status and larvae population numbers therein. All surface containers located within a 100 m radius of each well were removed, treated with s-methoprene or sealed to prevent the utilization of these containers by mosquitoes. These inner cores were surrounded by outer zones for a further 100 m in which surface containers were left untreated but all subterranean habitats were treated. Ovitraps were monitored monthly in the inner cores for 36 wk from August 1999 to April 2000 and differences in the proportions of ovitraps positive for Ae. aegypti and Ochlerotatus notoscriptus (Skuse) were analyzed by logistic regression. Analysis of the proportions of ovitraps positive for Ae. aegypti near positive wells indicated significantly greater colonization from November to March (the wet season), compared with those situated near Ae. aegypti negative wells. As Oc. notoscriptus were not produced from subterranean sites, comparisons of the proportions of ovitraps positive for Oc. notoscriptus in positive and negative inner cores provided an indication of the relative productivity of the uncontrolled surface containers in the outer zones. Differences in the utililization of ovitraps by Oc. notoscriptus among positive and negative cores were observed during only one month (March), when oviposition was greater in ovitraps in the negative cores, compared with the positive cores. Best subsets linear regression analysis of the proportion of ovitraps positive for Ae. aegypti against meteorological variables (rainfall, mean wind speed, mean relative humidity, mean minimum, and maximum temperature) during the week of ovitrapping indicated that minimum temperature and wind speed accounted for 63.4% of the variability. This study confirms that for semiarid towns such as Charters Towers, the practice of treating a relatively small number of key subterranean habitats during winter will significantly affect Ae. aegypti recolonization of surface container habitats during summer, the period of greatest risk for dengue.

Structure of the gut contents of Trichogramma australicum Girault (Hymenoptera : Trichogrammatidae) larvae fixed in situ in eggs of its host Helicoverpa armigera (Hubner) (Lepidoptera : Noctuidae)

Trichogramma australicum larvae develop most rapidly in younger eggs of its host, the pest lepidopteran Helicoverpa armigera . To establish how the developmental stage of the host affects the diet of T. australicum , larvae were fixed in situ in eggs of H. armigera of different ages and the structure of the egg contents and parasitoid gut contents examined histologically. Larvae feeding on newly laid host eggs contain primarily yolk particles in their gut, while larvae feeding on older hosts contain necrotic cells and yolk particles. The gut of T. australicum larvae does not contain organised tissue remnants, indicating that larvae feed primarily by sucking food into their pharynx and feed best on a mixture of particulate semisolids in a liquid matrix. Secretory structures of T. australicum larvae that could be involved in modifying the host environment were examined. The hindgut is modified to form an anal vesicle with a number of attributes suggesting that it may be a specialised secretory structure. The paired salivary glands open to the exterior via a common duct.

Susceptibility of Aedes aegypti larvae to temephos and Bacillus thuringiensis var israelensis in integrated control

The susceptibility of field collected Aedes aegypti larvae was evaluated in terms of median lethal time (LT50) and final mortality, when treated with temephos, Bacillus thuringiensis var israelensis as well as mixtures of these two agents. Third instar larvae were shown to be more susceptible than early and late fourth instar ones to the entomopathogen. Survival of some individuals when exposed to temephos suggest possible resistance. Temporal synergism in early fourth instar larvae was detected when they were exposed to mixtures of Bti-temephos. The possibility of this integrated treatment is commented on.