Superparasitism and Population Regulation of the Mosquito-Parasitic Mermithid Nematodes Romanomermis iyengari and Strelkovimermis spiculatus

Superparasitism is a common phenomenon in mosquito-parasitic mermithid nematodes. Multiple nematodes are needed in a single host to produce males. Host selection behavior and intraspecific competition among Romanomermis iyengari and Strelkovimermis spiculatus were investigated against their host, Culex pipiens pipiens in laboratory experiments. In a choice assay between previously infected and uninfected host larvae, infectious preparasites of both nematode species could distinguish not only between infected and uninfected hosts, but even between different parasite loads in showing a strong preference for uninfected hosts or hosts with a low parasite load. Host heart rate declined briefly immediately after parasitism. Superparasitism resulted in increased parasite mortality. Scramble competition within mosquito larvae for limited host nutrients, coupled with a skewed sex ratio favoring males, is assumed to lead to parasite population decline and subsequently toward host-parasite population equilibrium. The ability of mermithid preparasites to accurately assess parasite load likely plays an important role in host population dynamics and regulation

Behavioral differences of invasive container-dwelling mosquitoes to a native predator

ABSTRACT Aquatic prey show behavioral modiÞcations in the presence of predation-risk cues that alleviate their risk from predation. Aedes albopictus (Skuse), Aedes japonicus (Theobald), and Culex pipiens L. are invasive mosquitoes in North America, and their larvae are prey for the native mosquito predator, Toxorhynchites rutilus (Coquillett). Ae. albopictus and Ae. japonicus are recent invaders, whereas Cx. pipiens has been in the United States for Ͼ100 yr. In the presence of predation-risk cues from Tx. rutilus larvae, Cx. pipiens larvae increased the time spent resting at the surface (least risky behavior) more than the other prey species. Ae. japonicus larvae increased resting at the surface of the containers more than Ae. albopictus larvae in the presence of predation-risk cues. Cx. pipiens larvae spent more time motionless at the surface even in the absence of predation-risk cues when compared with the other species, indicating that Cx. pipiens larvae are the least vulnerable prey. As compared with the other prey species, Ae. albopictus larvae exhibited more high-risk behaviors both in the presence and absence of predation-risk cues, indicating that they are the most vulnerable prey. Ae. albopictus is the superior competitor; however, predation by Tx. rutilus larvae may prevent competitive exclusion by Ae. albopictus and promote coexistence among the three prey species

Transcriptomic analysis of the entomopathogenic nematode Heterorhabditis bacteriophora TTO1

Background: The entomopathogenic nematode Heterorhabditis bacteriophora and its symbiotic bacterium, Photorhabdus luminescens, are important biological control agents of insect pests. This nematode-bacterium-insect association represents an emerging tripartite model for research on mutualistic and parasitic symbioses. Elucidation of mechanisms underlying these biological processes may serve as a foundation for improving the biological control potential of the nematode-bacterium complex. This large-scale expressed sequence tag (EST) analysis effort enables gene discovery and development of microsatellite markers. These ESTs will also aid in the annotation of the upcoming complete genome sequence of H. bacteriophora. Results: A total of 31,485 high quality ESTs were generated from cDNA libraries of the adult H. bacteriophora TTO1 strain. Cluster analysis revealed the presence of 3,051 contigs and 7,835 singletons, representing 10,886 distinct EST sequences. About 72% of the distinct EST sequences had significant matches (E value < 1e-5) to proteins in GenBank's non-redundant (nr) and Wormpep190 databases. We have identified 12 ESTs corresponding to 8 genes potentially involved in RNA interference, 22 ESTs corresponding to 14 genes potentially involved in dauer-related processes, and 51 ESTs corresponding to 27 genes potentially involved in defense and stress responses. Comparison to ESTs and proteins of free-living nematodes led to the identification of 554 parasitic nematode-specific ESTs in H. bacteriophora, among which are those encoding F-box-like/WD-repeat protein theromacin, Bax inhibitor-1-like protein, and PAZ domain containing protein. Gene Ontology terms were assigned to 6,685 of the 10,886 ESTs. A total of 168 microsatellite loci were identified with primers designable for 141 loci. Conclusion: A total of 10,886 distinct EST sequences were identified from adult H. bacteriophora cDNA libraries. BLAST searches revealed ESTs potentially involved in parasitism, RNA interference, defense responses, stress responses, and dauer-related processes. The putative microsatellite markers identified in H. bacteriophora ESTs will enable genetic mapping and population genetic studies. These genomic resources provide the material base necessary for genome annotation, microarray development, and in-depth gene functional analysis

Ovicidal activity of three insect growth regulators against Aedes and Culex mosquitoes

a b s t r a c t Interspecific variations in the susceptibility of freshly and embryonated eggs of Aedes albopictus, Ae. aegypti, Ae. atropalpus and Culex pipiens were tested against three classes of insect growth regulators (IGRs) including ecdysone agonist (azadirachtin), chitin synthesis inhibitor (diflubenzuron) and juvenile hormone analog (pyriproxyfen) at 0.001, 0.01, 0.1 and 1.0 ppm concentrations. Egg hatching inhibition was dose dependent, the highest being at 1.0 ppm concentration for freshly laid eggs of Ae. albopictus (pyriproxyfen: 80.6%, azadirachtin: 42.9% and diflubenzuron: 35.8%). Aedes aegypti showed lower egg hatching inhibition when exposed to pyriproxyfen (47.3%), azadirachtin (15.7%) and diflubenzuron (25.5%). Freshly laid eggs of Cx. pipiens were most susceptible to diflubenzuron. Aedes atropalpus eggs were tolerant to all three classes of IGRs. Embryonated eggs of Ae. albopictus, Ae. aegypti, Ae. atropalpus and Cx. pipiens were resistant to pyriproxyfen, azadirachtin and diflubenzuron than freshly laid eggs. The median desiccation time (DT 50 ) of Ae. atropalpus eggs was maximum (5.1 h) as compared to Ae. aegypti (4.9 h), Ae. albopictus (3.9 h) or Cx. pipiens (1.7 h) eggs. Insignificant relationship between the rates of desiccation and egg hatching inhibition suggests other factors than physical providing eggs the ability to tolerate exposures to various IGRs. Egg hatching inhibition was due to the alteration in embryonic development caused by IGRs. Changes in the egg shell morphology and abnormal egg hatching from the side of the egg wall instead of operculum, was observed at higher concentrations of diflubenzuron. Morphological and physiological variations in eggs may be the key factor to influence the ovicidal efficacy of IGRs. The present data provide a base line for the improvement of the ovicidal efficacy of the insecticide and its formulation

A Lover and a Fighter: The Genome Sequence of an Entomopathogenic Nematode Heterorhabditis bacteriophora

Heterorhabditis bacteriophora are entomopathogenic nematodes that have evolved a mutualism with Photorhabdus luminescens bacteria to function as highly virulent insect pathogens. The nematode provides a safe harbor for intestinal symbionts in soil and delivers the symbiotic bacteria into the insect blood. The symbiont provides virulence and toxins, metabolites essential for nematode reproduction, and antibiotic preservation of the insect cadaver. Approximately half of the 21,250 putative protein coding genes identified in the 77 Mbp high quality draft H. bacteriophora genome sequence were novel proteins of unknown function lacking homologs in Caenorhabditis elegans or any other sequenced organisms. Similarly, 317 of the 603 predicted secreted proteins are novel with unknown function in addition to 19 putative peptidases, 9 peptidase inhibitors and 7 C-type lectins that may function in interactions with insect hosts or bacterial symbionts. The 134 proteins contained mariner transposase domains, of which there are none in C. elegans, suggesting an invasion and expansion of mariner transposons in H. bacteriophora. Fewer Kyoto Encyclopedia of Genes and Genomes Orthologies in almost all metabolic categories were detected in the genome compared with 9 other sequenced nematode genomes, which may reflect dependence on the symbiont or insect host for these functions. The H. bacteriophora genome sequence will greatly facilitate genetics, genomics and evolutionary studies to gain fundamental knowledge of nematode parasitism and mutualism. It also elevates the utility of H. bacteriophora as a bridge species between vertebrate parasitic nematodes and the C. elegans model

Transcriptional profiling of trait deterioration in the insect pathogenic nematode Heterorhabditis bacteriophora

Background: The success of a biological control agent depends on key traits, particularly reproductive potential, environmental tolerance, and ability to be cultured. These traits can deteriorate rapidly when the biological control agent is reared in culture. Trait deterioration under laboratory conditions has been widely documented in the entomopathogenic nematode (EPN) Heterorhabditis bacteriophora (Hb) but the specific mechanisms behind these genetic processes remain unclear. This research investigates the molecular mechanisms of trait deterioration of two experimental lines of Hb, an inbred line (L5M) and its original parental line (OHB). We generated transcriptional profiles of two experimental lines of Hb, identified the differentially expressed genes (DEGs) and validated their differential expression in the deteriorated line. Results: An expression profiling study was performed between experimental lines L5M and OHB of Hb with probes for 15,220 ESTs from the Hb transcriptome. Microarray analysis showed 1,185 DEGs comprising of 469 down- and 716 up-regulated genes in trait deteriorated nematodes. Analysis of the DEGs showed that trait deterioration involves massive changes of the transcripts encoding enzymes involved in metabolism, signal transduction, virulence and longevity. We observed a pattern of reduced expression of enzymes related to primary metabolic processes and induced secondary metabolism. Expression of sixteen DEGs in trait deteriorated nematodes was validated by quantitative reverse transcription-PCR (qRT-PCR) which revealed similar expression kinetics for all the genes tested as shown by microarray. Conclusion: As the most closely related major entomopathogen to C. elegans, Hb provides an attractive near-term application for using a model organism to better understand interspecies interactions and to enhance our understanding of the mechanisms underlying trait deterioration in biological control agents. This information could also be used to improve the beneficial traits of biological control agents and better understand fundamental aspects of nematode parasitism and mutualism

Source Reduction Behavior as an Independent Measurement of the Impact of a Public Health Education Campaign in an Integrated Vector Management Program for the Asian Tiger Mosquito

The goal of this study was to evaluate the effectiveness of a public health educational campaign to reduce backyard mosquito-larval habitats. Three communities each, within two New Jersey counties, were randomly selected to receive: (1) both education and mosquito control, (2) education only, and (3) no education or mosquito control. Four separate educational events included a 5-day elementary school curriculum in the spring, and three door to door distributions of educational brochures. Before and after each educational event, the numbers of mosquito-larval container habitats were counted in 50 randomly selected homes per study area. Container surveys allowed us to measure source reduction behavior. Although we saw reductions in container habitats in sites receiving education, they were not significantly different from the control. Our results suggest that traditional passive means of public education, which were often considered the gold standard for mosquito control programs, are not sufficient to motivate residents to reduce backyard mosquito-larval habitats