Evidence for the copepods Acanthocyclops robustus and Mesocyclops edax as competent intermediate hosts for Coelomomyces punctatus during an epizootic in a larval population of the mosquito Anopheles quadrimaculatus.

Field and laboratory experiments were conducted during an epizootic of Coelomomyces punctatus (Chytridiomycetes: Blastocladiales) in a population of the mosquito Anopheles quadrimaculatus in a North Carolina farm pond to examine the interactions of several potential copepod hosts with the mosquito and fungus. The diel vertical migratory behavior of the copepod species Acanthocyclops robustus, Eucyclops serrulatus, Macrocyclops albidus, and Mesocyclops edax were monitored in relation to infection rates in sentinel mosquito larvae. Mosquito infection occurred primarily around dusk, the same period during which A. robustus and E. serrulatus were most abundant near the surface of the pond. However, exposure of A. robustus, E. serrulatus, M. albidus, M. edax, Microcyclops varicans, and Paracyclops poppei to fungal meiospores in the laboratory showed that only A. robustus and M. edax were competent intermediate hosts for C. punctatus. Laboratory studies of the diel periodicity of gametangial dehiscence in A. robustus and M. edax infected with C. punctatus revealed that gamete release and zygote formation also occurred around dusk. The combined results of the laboratory and field studies on copepod abundance, susceptibility to infection, and periodicity of gametangial dehiscence suggest that A. robustus was the principal intermediate host for C. punctatus during the epizootic, though it is probable that M. edax also contributed importantly to the overall rate of larval infection.     

Sustainable control of mosquito larvae in the field by the combined actions of the biological insecticide Bti and natural competitors

Integrated management of mosquitoes is becoming increasingly important, particularly in relation to avoiding recolonization of ponds after larvicide treatment. We conducted for the first time field experiments that involved exposing natural populations of the mosquito species Culex pipiens to: a) application of the biological insecticide Bacillus thuringiensis israelensis (Bti), b) the introduction of natural competitors (a crustacean community composed mainly of Daphnia spp.), or c) a combined treatment that involved both introduction of a crustacean community and the application of Bti. The treatment that involved only the introduction of crustaceans had no significant effect on mosquito larval populations, while treatment with Bti alone caused only a significant reduction in the abundance of mosquito larvae in the short‐term (within 3–10 days after treatment). In contrast, the combined treatment rapidly reduced the abundance of mosquito larvae, which remained low throughout the entire observation period of 28 days. Growth of the introduced crustacean communities was favored by the immediate reduction in the abundance of mosquito larvae following Bti administration, thus preventing recolonization of ponds by mosquito larvae at the late period (days 14–28 after treatment). Both competition and the temporal order of establishment of different species are hence important mechanisms for efficient and sustainable mosquito control.     

Effects of dairy wastewater on mosquitoes in southern California.

The effect of dairy pond water quality on mosquito biology was studied under laboratory conditions during 2004. Due to high turbidity and larval toxicity of raw water samples from dairy ponds with no mosquito breeding, water samples from both mosquito breeding and non-breeding ponds were filtered and diluted 5X with distilled water prior to being tested for sublethal effects on mosquito development. The diluted pond waters showed no significant deleterious effects on the overall development of Culex quiquefasciatus Say at 23.9 degrees and 26.6 degrees C. However, the 3rd and 4th instar larvae were significantly shorter and lighter in weight when reared in diluted water samples from mosquito non-breeding ponds. Similar effects were found on the developmental stages of Cx. tarsalis Coquillette. Data on water quality parameters of filtered pond waters showed higher levels of hexavalent chromium (4X), nitrites (10X), nitrates (2X), sulfates (5X), and salinity (2X) in water samples from non-breeding ponds. Based on these data, the presence of high levels of hexavalent chromium and sulfate appear to be responsible for the absence of mosquito breeding in some dairy ponds.     

Vertical transmission of Nosema fumiferanae (Microsporidia: Nosematidae) and consequences for distribution, post-diapause emergence and dispersal of second-instar larvae of the spruce budworm, Choristoneura fumiferana (Clem.) (Lepidoptera: Tortricidae)

We examined vertical transmission of Nosema fumiferanae in the eastern spruce budworm, Choristoneura fumiferana (Clem.) (Lepidoptera: Tortricidae), and how it affects overwintering distribution and survival and spring emergence and dispersal of second-instar larvae in outbreak populations. Females containing 5.0 x 10(5) spores or more consistently produced 100% infected progeny. Transmission efficiency was still 50% at burdens as low as 0.2 x 10(5) spores per moth. Infection intensity in offspring increased with maternal spore load but became highly variable above 25 x 10(5) spores per female. Nosema multiplied in second instars for at least 1 month after they entered dormancy, regardless of temperature (2 degrees C versus 21 degrees C). Infection did not affect the distribution of overwintering larvae in a white spruce canopy. Dormancy survival between late-summer and the following spring was lower in families from infected females and was negatively correlated with larval infection intensity. Infection delayed larval emergence from hibernacula in the spring and resulted in delayed dispersal of emerged larvae, at least when parasite prevalence and infection intensities were high. Infected larvae were less successful in establishing feeding sites after dispersal. Our results underscore the potential of Nosema infection to negatively affect processes early in the budworm life cycle.     

Differential crop damage by healthy and nucleopolyhedrovirus-infected Mamestra brassicae L. (Lepidoptera: Noctuidae) larvae: a field examination

Baculovirus infection in Lepidoptera can alter both larval mobility and feeding rates, which can in turn affect pathogen transmission and dispersal in the field. We compared the damage to cabbage plants in the field caused by healthy and nucleopolyhedrovirus-infected Mamestra brassicae L. (Lepidoptera: Noctuidae) larvae released as second and fourth instars. There was no significant difference in plant consumption by healthy and infected larvae for the first 4 days after release. From day 5 onwards, infected larvae caused significantly less defoliation. This pattern was similar for larvae at both larval instars. Defoliation was greater for fourth instars throughout the experiment.     

Ecological aspects between Contracaecum sp. (Nematoda, Anisakidae) and the host Serrasalmus spilopleura Kner, 1860 (Pisces, Characidae) in natural populations of northeastern Argentina

From February 1987 to February 1989, the populational biology of Contraceacum sp. (larvae) in its paratenic host, the fish Serrasalmus spilopleura Kner, 1860, was studied in two ponds in a subtropical permanent habitat northeastern of Argentina. Fishes from Ramada Paso pond presented 80% of prevalence and 1 to 132 larvae per fish while fishes from Aeroclub pond presented 63% of prevalence and 1 to 184 larvae per fish. Fishes collected from Aeroclub pond have shown a high prevalence of infection during the first period of study (1987), diminishing the following year. In fishes from Ramada Paso pond the prevalence varied not significatively during the two years. Prevalence and mean intensity of infection increase with body length and weight of the hosts. Sex of hosts is not an influential factor in parasitic level. The lenitic "closed" environmental (Ramada Paso pond) evidenced the greatest larvae mean intensity and prevalence. Although, the lenitic "open" environmental (Aeroclub pond) showed the greatest parasitic number of individuals in an infrapopulation. The spatial dispersion in both ponds were aggregated and fit well a negative binomial model. Nevertheless, the Aeroclub pond presented the greatest overdispersion.     

Larval development and growth ratios of Odonata of the Azores

To fully understand odonate life cycles, it is vital to analyse the patterns of larval growth, which are tightly associated with variations in environmental factors. However, the identification of larval instars is often difficult, especially for early development stages. We hypothesise that pond hydroperiod influences odonate larval growth, and test this idea with novel information about the environmental characteristics of 12 study ponds in Terceira Island (Azores). This study, which represents the first analysis of larval development of Odonata in the Azores, involved determining the instars of development and growth ratios. We measured the morphological characteristics of 898 Odonata larvae, and found that they significantly differed between temporary and permanent ponds. To estimate the larval instars, we used two methods: frequency distribution methods and correlation diagrams between body characteristics. The first method was the most effective, allowing the estimation of 17 instars for Anax imperator, 13 for Sympetrum fonscolombii and 10 for Ischnura hastata, as well as growth ratios between instars for head width, total length, abdomen length, wings and antenna length. Our results also suggest that a combination of the two methods is the most appropriate strategy for estimating the number of instars and growth ratios during larval development.     

Experimental laboratory infection of mosquito larvae with fungi of the genus Coelomomyces. II. Experiments with Coelomomyces punctatus in Anopheles quadrimaculatus

Experiments on the infection of Anopheles quadrimaculatus with Coelomomyces punctatus indicate that planonts released from sporangia are not infective for mosquito larvae but most likely infect the copepod Cyclops vernalis. Exposure of early-instar larvae to up to 5 × 10³ planonts per larva for as long as 48 hr resulted in no larval infections. Motile planonts were no longer detectable after 48 hr. However, incubation of early-instar larvae in media to which planonts, algae, and copepods had been added several days previously resulted in larval infection. Infection did not occur during the first 6 days after planont introduction. On day 7 and for several days thereafter, copepods were detected in the media which had an extensive mycelium developing in the hemocoel. This mycelium cleaved into thousands of posteriorly uniflagellate planonts. The presence of planonts at the time of mosquito infection, in conjunction with the above results, suggests that Cyclops vernalis is an alternate host for Coelomomyces punctatus and that the latter has a life cycle similar to that proposed for C. psorophorae involving a mosquito and a copepod as obligate alternate hosts. In established infection containers, dilution of the media with water significantly increased levels of infection 6 days later. All larval instars were susceptible to C. punctatus.     

Factors influencing the abundance of Japanese encephalitis vectors in ricefields in India--II. Biotic

The relationship of insect predators and phytoplankton with the abundance of Culex tritaeniorhynchus Giles, Cx. vishnui Theobald and Cx. pseudovishnui Colless mosquito larvae and pupae (Diptera: Culicidae) in ricefields was investigated during three rice growing seasons. Notonectids were the most abundant insect predators, whereas diatoms dominated among phytoplankton. Multiple regression analysis showed that the occurrence of notonectids (both nymphs and adults) was negatively associated with larval abundance. Phytoplankton, especially diatoms and blue green algae (BGA), were found to favour abundance of culicine immatures during Navarai and Kuruvai crops, respectively. Larval gut analysis showed that the intake of algae by late instars was high, with 93%, 58% and 24% of diatoms, BGA and green algae, respectively. Filamentous algae may not necessarily be of nutritive value, but they are observed to form mats, which provide protection to the mosquito immatures from the predators.     

Parasitoid-induced behavioral alterations of Aedes aegypti mosquito larvae infected with mermithid nematodes (Nematoda: Mermithidae).

A wide range of parasites are known to cause behavioral changes in their hosts and parasitized insects are especially amenable to the study of such changes. The majority of studies addressing parasite-induced behavioral alterations have focused on parasites with complex life cycles and the adaptive nature of such changes. Behavioral changes caused by parasitoids, single-host parasites that kill their host upon emergence, have been studied less and the adaptive nature of these changes is likely to be different than those in complex life cycles. I investigated behavioral alterations in Aedes aegypti mosquito larvae infected with parasitoid nematodes (family Mermithidae). I conducted several experiments in which I tested the following hypotheses: 1) Mermithid nematodes induce behavioral changes in mosquito larvae and the changes are density dependent. 2) Different species of mermithid nematodes induce similar changes in mosquito larvae behavior. 3) Behavioral alterations vary with mermithid developmental stage. 4) Mosquito larvae infected with mermithid nematodes behave similarly to uninfected food-deprived mosquito larvae. I found that 4th instar Ae. aegypti infected with Romanomermis culicivorax or Strelkovimermis spiculatus exhibited resting behaviors significantly more often than uninfected controls but that intensity of infection did not affect activity levels. In earlier instars, infected mosquito larvae were more active than uninfected control larvae in some behaviors associated with feeding. There was no significant difference between infected and uninfected food-deprived mosquitoes in nine of the ten behaviors observed. The decrease in activity of late instar Ae. aegypti larvae infected with mermithids may be a parasitoid adaptation that reduces the risk of predation and thus increases host and parasitoid survival. The increase in feeding activity in earlier instars as well as the similarity between uninfected food-deprived and infected Ae. aegypti behavior may indicate that these behaviors are adaptive for the parasitoid, increasing nutritional acquisition for successful parasitoid development.     

Crustacean biodiversity as an important factor for mosquito larval control

Newly established ponds, which are highly dynamic systems with changing levels of biological interactions among species, are common larval mosquito habitats. We investigated the impact of crustacean abundance and taxa diversity on mosquito oviposition and larval development. The effects of the biological larvicide Bacillus thuringiensis israelensis (Bti) on mosquito larvae were monitored according to fluctuations in crustacean communities. Populations of the mosquito Culex pipiens colonized artificial ponds that contained crustacean communities at different time points of colonization by crustaceans: 1) ‘no colonization’ (no crustaceans), 2) ‘simultaneous colonization’ by crustaceans and mosquitoes, and 3) ‘head‐start colonization’ by crustaceans (preceding colonization by mosquitoes). All types of ponds were treated with three concentrations of Bti (10, 100, or 1,000 µg/liter). Colonization of all ponds by Cx. pipiens (in terms of oviposition, larval abundance, and larval development) decreased significantly with increasing diversity of crustacean taxa. The total abundance of crustaceans had a minor effect on colonization by Cx. pipiens. The presence of crustaceans increased the sensitivity of Cx. pipiens larvae to Bti treatment by a factor of 10 and delayed the time of recolonization. This effect of Bti was relevant in the short term. In the long term, the presence of Cx. pipiens was determined by crustacean biodiversity.     

Effect of entomopoxvirus infection of the smaller tea tortrix, Adoxophyes sp. on the development of the endoparasitoid, Ascogaster reticulatus

Infection of Adoxophyes sp. (Lepidoptera: Tortricidae) larvae by an entomopoxvirus (AsEPV) adversely affected the development of the endoparasitoid, Ascogaster reticulatus Watanabe (Hymenoptera: Braconidae). Parasitoid larvae developing in AsEPV-infected hosts grew more slowly and spent more time in their hosts than did parasitoid larvae developing in noninfected hosts. Percentages of emergence of larval parasitoids that developed in AsEPV-infected hosts were significantly lower than those of parasitoids that developed in noninfected hosts. Parasitoid larvae in AsEPV-infected host perished when their hosts died of AsEPV infection. Significant numbers of parasitized and infected larvae exhibited apolysis to the final instar, whereas noninfected-parasitized larvae died in the penultimate instars due to emergence of parasitoids.     

Biological Control of Culex pipiens pattens (Diptera, Culicidae) by the Release of Fish Muddy Loach, Misgurnus mizolepis in Natural Ponds, Korea

ABSTRACT An assessment on the biological control potential with the fish muddy loaches (Misgurnus mizolepis) was conducted against naturally bred Culex pipiens pollens larvae in four ponds (A, B, C and D) in Busan from July through September, 2001. Predation of the fish at 3 different release rates of 4,5, and 6 fish/m² resulted in mostly over 90% mosquito control from the first week after treatment through the end of the survey period for 11 weeks. There were no significant difference among the release rates of fish at the 5% level of probability. However, substantial controls of 43.0% and 25.9% were obtained from pond C during the 3rd and 7th weeks after the fish introduction, respectively. The results of those two weeks showed a lower biological control by the introduced larvivorous fishes. This might be due to the presence of heavy organic matters including aquatic weeds and/or severely polluted water from sewage in pond C. The aquatic weeds covered the pond's water surface which may have affected the deterioration of mosquito preying in favor of aquatic weeds. Also, the fishes were observed to avoid severely contaminated sewage water in some parts of ponds A and C where more mosquito larvae were found.     

马尾松毛虫过氧化氢酶及过氧化物酶与耐药性的关系

马尾松毛虫Dendrolimus punctatus幼虫体内存在着过氧化氢酶(CAT)和过氧化物酶(POD)。4龄幼虫的CAT和POD活力较大,其次是6龄幼虫,5龄幼虫的CAT和POD活力较4龄和6龄幼虫低。醚菊酯(etofenprox)处理后,在兴奋期(30 min),CAT和POD活力水平上升。4龄和6龄幼虫在抑制期(50 min以后),CAT和POD活力呈波动式上升,接近死亡时下降。5龄幼虫的CAT和POD活力呈波动式上升,接近死亡时下降。5龄幼虫的CAT在抑制期保持比正常虫体高的活力。结果表明,马尾松毛虫4龄、5龄和6龄幼虫与耐药性存在一定的相关性,研制酶的抑制剂具有实用意义。根据毒力测定结果,马尾松毛虫幼虫对醚菊酯的耐药力,5龄是4龄的1.43倍,6龄是4龄的1.72倍。因此,药物防治的合理时期应掌握在4龄以前较适宜。     

Physiological and morphological aspects of Aedes aegypti developing larvae: effects of the chitin synthesis inhibitor novaluron

Population control of the dengue vector mosquito, Aedes aegypti, is difficult due to many reasons, one being the development of resistance to neurotoxic insecticides employed. The biosynthesis of chitin, a major constituent of insect cuticle, is a novel target for population control. Novaluron is a benzoylphenylurea (BPU) that acts as a chitin synthesis inhibitor, already used against mosquitoes. However, information regarding BPU effects on immature mosquito stages and physiological parameters related with mosquito larval development are scarce. A set of physiological parameters were recorded in control developing larvae and novaluron was administered continuously to Ae. aegypti larvae, since early third instar. Larval instar period duration was recorded from third instar until pupation. Chitin content was measured during third and fourth instars. Fourth instars were processed histochemically at the mesothorax region, stained with hematoxylin and eosin (HE) for assessment of internal tissues, and labeled with WGA-FITC to reveal chitinized structures. In control larvae: i) there is a chitin content increase during both third and fourth instars where late third instars contain more chitin than early fourth instars; ii) thoracic organs and a continuous cuticle, closely associated with the underlying epidermis were observed; iii) chitin was continuously present throughout integument cuticle. Novaluron treatment inhibited adult emergence, induced immature mortality, altered adult sex ratio and caused delay in larval development. Moreover, novaluron: i) significantly affected chitin content during larval development; ii) induced a discontinuous and altered cuticle in some regions while epidermis was often thinner or missing; iii) rendered chitin cuticle presence discontinuous and less evident. In both control and novaluron larvae, chitin was present in the peritrophic matrix. This study showed quantitatively and qualitatively evidences of novaluron effects on Ae. aegypti larval development. To our knowledge, this is the first report describing histological alterations produced by a BPU in immature vector mosquitoes.     

Instar Susceptibility of Simulium vittatum (Diptera: Simuliidae) to the Entomogenous Nematode Neoaplectana carpocapsae

Laboratory bioassays showed that the susceptibility of Simulium vittatum to Neoaplectana carpocapsae increased with successive larval instars. First, second, and third instar larvae were resistant to infection, while seventh instars were highly susceptible. Significant differences in intra-instar susceptibility were also evident, as mortality ranged from 58% for the smallest seventh instar larvae to 97% for the largest. Dissections revealed that the basis for the resistance of early instars was physical exclusion of the comparatively large nematodes. The principle factor regulating the susceptibility of mid and late instars was injury to nematodes caused by larval mouthparts during ingestion. Differences in intra-instar susceptibility were similarly related to nematode injury.     

Do nymphs of Parameletus chelifer and P. minor (Ephemeroptera) reduce mortality from predation by occupying temporary habitats?

Summary The possible effect of predation on the habitat selection of nymphs of the two mayfly species Parameletus chelifer Bengtsson and P. minor (Bengtsson) was investigated for different habitats of a North Swedish boreal river. In spring, these two species migrate to river margins and subsequently, in different proportions, to adjacent seasonal ponds. Feeding rates on mayfly nymphs and mosquito larvae by some dytiscid beetles and larvae, a corixid waterbug and a caddisfly larva were studied in the laboratory. In the seasonal pond, the consumption rate of invertebrate predators on Parameletus nymphs was below that expected from their relative abundance, and mosquito larvae were preferred. At the river margin, where mosquito larvae were absent, Parameletus nymphs were consumed in agreement with their relative abundance. The absolute abundance of macroinvertebrate predators was similar at the river margin and in the pond. Fish predators visited the seasonal pond only during a short time of flood, while at the river margin their presence was continuous. In the seasonal pond, minnow and roach consumed nymphs of both P. chelifer and P. minor in a lower proportion than expected. At the river margin, nymphs of P. chelifer were consumed by minnow and ten-spined stickleback in a higher proportion than expected. Nymphs of P. minor were consumed according to expectance. Imagines of P. minor emerged early, and nymphs were equally abundant in the river and in the seasonal pond. In contrast, almost all nymphs of P. chelifer migrated to the seasonal pond. The later emergence of P. chelifer would probably result in a very high mortality from predation if it remained at the river margin.     

Studies on the biology of Chaoborus flavicans (Meigen) (Diptera: Chaoboridae) in a fish-free eutrophic pond,Japan

Population dynamics of Chaoborus flavicans larvae of various instars was studied from November 1986 to December 1987 in a eutrophic, fish-free pond, Japan. First and 2nd instar larvae were observed from late April to late October, indicating a reproductive period of about half a year. C. flavicans overwintered in the 4th instar larvae. In water column samples, total density of all instars was 680–23680 m^-2, and pupal density 0–2600 m^-2; larvae of the 1st, 2nd, and 3rd instars showed 5–6 density peaks in 1987, suggesting that 5–6 generations occur during a year (peaks of the 4th instar larvae were not clear, probably due to their longer development than those of younger instars). In sediment samples, no 1st and 2nd instar larvae were found, 3rd instar larvae were found occasionally but density of the 4th instar larvae was 280–18600 m^-2, and pupal density varied between 0–502 m^-2. Fouth instar larvae accumulated in sediment in the cold season and in the water column in the warm season; high temperature and low oxygen concentration were the most important factors limiting the distribution of larvae in the sediment in summer in the NIES pond. The dry weight of total C. flavicans larvae was 0.08–4.2 g m^-2 in sediment samples and 24–599 μg l^-1 (0.10–2.40 g m^-2) in water column samples. Comparisons of maximum densities in the NIES pond in different years and in waters of different trophic status show that density is generally higher in eutrophic than in oligotrophic habitats. This revised version was published online in July 2006 with corrections to the Cover Date.     

Modified Behavior in Baculovirus-Infected Lepidopteran Larvae and Its Impact on the Spatial Distribution of Inoculum

Quantifying the rate of dispersal of target insects when infected with a disease agent will aid the development of biorational pest control programs. The effect of nucleopolyhedrovirus (NPV) infection on the mobility of second and fourth instarMamestra brassicaelarvae was investigated in the laboratory and field. NPV infection altered larval mobility, with the changes in behavior varying with the timecourse of infection. Diseased larvae moved three to five times further than healthy ones during the middle stages of infection. By the 7th day postinfection diseased larvae were less mobile than healthy counterparts. The same pattern of modified behavior was observed in both instars. Fourth instar larvae moved further than second instars under laboratory and field conditions. In the field, infected larvae tended to die on the apex of the cabbage leaves. Bioassay of the leaves showed a linear decrease in inoculum from central to peripheral plants within the plots, which occurred to the same extent for second and fourth instars. Leaves from plots where infected fourth instar larvae had been introduced had higher inoculum density than those from plots with second instars.