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.     

Effect of larval maturation on mortality induced by nuclear polyhedrosis and granulosis virus infections of Heliothis armigera

All the instars of Heliothis armigera larvae were found to be susceptible to both nuclear polyhedrosis virus (NPV) and granulosis virus (GV). An inverse relationship between mortality and larval age was found in the case of the NPV, while the GV displayed a rather erratic mortality pattern. A degree of maturation immunity against the NPV was found to exist, but the same is not true for the GV. The important role that pupation plays on the effect of a lethal infection is also discussed.     

Overwintering,soil distribution and phenology of Childers canegrub,Antitrogus parvulus (Coleoptera: Scarabaeidae) in Queensland sugarcane

In this study, the question of whether Childers canegrub, Antitrogus parvulus (Britton) overwinters in the subsoil was addressed. Irrigated fields of sugarcane were sampled during a 2-year period near Bundaberg in southern Queensland. Antitrogus parvulus overwintered as second and third instars at each of three sites. During autumn and winter third instars of different allochronic (separated in age by 12 months) populations occurred together and could not be readily separated. Field-collected third instars were reared on ryegrass and separated into two age groups based on the date of pupation. Third instars in the first year of their life cycle (young third instars) remained at shallow depth (100-200 mm) and did not overwinter in the subsoil as once thought. Minimum temperatures during winter were 13-16 degrees C and did not prevent young third instars from feeding and gaining weight. Third instars in their second and final year moved downwards from late summer and pupated in the subsoil at 293-425 mm in spring. General phenology was as previously reported with first instar larvae occurring from January until April, second instars from January until November and third instar larvae throughout the year. Prepupae and pupae were found between October and December and adults occurred in soil during November and January. Batches of eggs occurred at a mean depth of 350 mm. First and second instars occurred predominantly at relatively shallow (100-200 mm) depths in the soil profile. All stages tended to be most common under rows of sugarcane rather than in the interrow.     

Stressors and rearing diseases of Trichoplusia ni: evidence of vertical transmission of NPV and CPV.

Inability to consistently rear healthy Trichoplusia ni led to a study of its rearing diseases. Four diseases were designated after preliminary research which included electron microscopy: cytoplasmic polyhedrosis (due to cytoplasmic polyhedrosis virus, or CPV), nuclear polyhedrosis (due to nucleopolyhedrovirus, or NPV), "neonate death" syndrome (mortality in first or second instars), and "late-instar" syndrome (death in late instars accompanied by bacterial decomposition). Infectious agents were not detected by electron microscopy in insects with the latter two diseases. Prevalence of CPV and NPV, but not the neonate-death or late-instar diseases, in progeny was significantly associated with pairs of mating adults. In conjunction with egg-surface decontamination, this indicated that both viruses may be transmitted transovarially. Pupae killed by CPV had virions in the cytoplasm of infected cells, but polyhedra were empty, not occluding virions. None of the diseases had a consistent pattern of prevalence associated with the date on which eggs were laid after oviposition began. Prevalence rates of cytoplasmic polyhedrosis, nuclear polyhedrosis, and the late-instar disease were significantly greater at 95-100% relative humidity (RH) than at RH levels of 75% or below. These same three diseases killed significantly more insects in crowded rearing conditions (four or five larvae per cup with 10.2-cm(2) diet surface) than in uncrowded conditions (one to three larvae per cup). As a result of these experiments, healthy T. ni have been reared for 10 generations by use of a modified Pasteur method and rearing cups containing no more than two larvae.     

Field evaluation of a DNA hybridization assay for nuclear polyhedrosis virus in gypsy moth (Lepidoptera: Lymantriidae) larvae.

DNA hybridization assays were used to detect the presence of viral DNA in gypsy moth (Lymantria dispar L.) larvae collected weekly from high density populations or reared from field-collected egg masses. DNA was extracted from larvae, bound to nitrocellulose filters, and hybridized with digoxigenin-labeled L. dispar NPV (LdNPV) DNA probes. The virus incidence determined from DNA hybridization assays was compared with that determined with conventional microscopic examination of larvae for polyhedral inclusion bodies. Among neonates reared from field-collected egg masses, average mortality from LdNPV (15.4%) within 10 d after hatch was not significantly different from the percentage of extracts containing LdNPV DNA (14.8%) found among larvae frozen 5 d after hatch before any mortality occurred. Field-collected larvae were split into two groups: half were frozen immediately and probed for LdNPV DNA and the other half were reared on artificial diet. The proportion containing LdNPV DNA closely approximated the proportion that died within 6 d of collection, but the proportion that died within 13 d of collection was underestimated.     

穴蚁蛉的自然种群动态

本文主要报道了穴蚁蛉 Myrmeleon(Morter) sagax(Walker)幼虫的野外自然种群数量动态。穴蚁蛉 1年发生 1代 ,在自然界其幼虫 3～ 5月份数量平稳略有下降 ,5、6月间数量迅速下降 ,6月初数量最少 ;7～ 9月份数量迅速上升 ,而后下降 ;新一代穴蚁蛉幼虫 (蚁狮 )在 6月份出现。其种群 ,秋季以 1龄幼虫数量为多 ,冬季主要以 2龄幼虫为主越冬 ,春季以 3龄蚁狮占优势。春季野外采集来的蚁狮 ,在每日光照 14小时、每周两饲 (每次饲 1头米蛾 Corcyra cephalonica成虫 )的条件下饲养 ,结果发现 ,采集回时处于 3龄期的蚁狮比自然界大约提前 1个月结茧化蛹和羽化 ;而采集回时处于 2龄期的蚁狮则和自然界中的情况基本一致。     

Effect of MON810 Bt field corn on Helicoverpa zea (Lepidoptera: Noctuidae) cannibalism and its implications to resistance development

Pairs of Helicoverpa zea (Boddie) larvae reared on diet-incorporated MON810 transgenic leaf tissue of field corn (Zea mays L.) were observed in the laboratory to characterize effects of sublethal levels of Bacillus thuringiensis variety kurstaki (Bt) Cry1Ab endotoxins on cannibalistic behavior and mortality. Feeding on sublethal levels of Bt corn reduced the frequency of cannibalistic behaviors exhibited by H. zea when uneven instars were paired together. Exposure to the Bt endotoxin had no significant effect on when cannibalistic mortality occurred or the level of mortality as a result of cannibalism. Assuming that H. zea larvae reared on nonBt corn tissue behaved in a similar way that resistant larvae would if feeding on Bt tissue, sublethal effects of Cry1Ab intoxication may reduce the chances of successful cannibalism by susceptible larvae and thus play a disproportionate role in the survival of multiple ear infestations. Furthermore, cannibalistic encounters could result in partially resistant larvae feeding on nontoxic food, thus temporarily providing an escape from exposure to the Bt endotoxin. These behavior alterations could increase the selective differential between susceptible individuals and those carrying resistance genes.     

Changes observed in hemolymph proteins of the lawn armyworm,Spodoptera mauritia acronyctoides,during growth,development, and exposures to a nuclear polyhedrosis virus

Vertical electrophoresis with acrylamide gel was used to study the effects of a nuclear polyhedrosis virus (NPV) on the hemolymph proteins of Spodoptera mauritia acronyctoides. An electrophoretic pattern consisting of 20 basic bands of proteins was separated in hemolymph of normal larvae which were older than 17 days. These hemolymph proteins increased quantitatively during growth. All 20 proteins could not be detected in hemolymph of younger larvae by the techniques utilized. Additional proteins were separated with metamorphosis. Lethal doses of NPV resulted in a general reduction of hemolymph proteins (hypoproteinemia) in infected larvae. Sublethal doses of NPV elicited an increase in certain hemolymph proteins. Similar increases in proteins were also observed in larvae surviving ostensibly lethal levels of NPV, in larvae subjected to physical stress, and in larvae reared axenically without formaldehyde in their diets. These same proteins, however, were present in approximately the same quantities in mature larvae. Physiopathology of NPV in S. mauritia appears to involve stress factors, host reactions, and the host endocrine system.     

Effect of Aedes aegypti (Diptera:Culicidae) Age on Sex Ratios in Romanomermis culicivorax (Nematoda:Mermithidae)

First, second, third, early and late fourth-instar larvae, and pupae of Aedes aegypti were infected with Romanomerrais culicivorax and reared at 20, 25, and 30 C. An increase in the ratio of male to female nematodes was observed with increase in host age at the time of infection at each temperature. The number of pupal and late fourth-instar infections was low, but R. culicivorax continued to develop in adult A. aegypti. Since male nematodes were recovered from both male and female hosts infected as late fourth instars or pupae, the sex of the host did not influence the sex of the nematode.     

Adaptations to captive breeding of the longhorn beetle Morimus funereus (Coleoptera: Cerambycidae); application on amylase study

Captive breeding has been suggested as a method of conservation for many vertebrates, and is increasingly being proposed as a strategy for invertebrates. In this study, the growth, development and fertility of adults of the vulnerable cerambycid Morimus funereus reared in captivity are examined. Two oviposition cycles; from May to September and from January to March were studied and larvae from wild adults and from the progeny of captive adults (second generation larvae) were examined. Five to 12 instars were observed during larval development. Larval development was completed in 218 days (average) for the progeny of wild adults with an average mortality rate of 10.3% and in 226 days (average) for larvae from captive adults with mortality rate of 34.9%. First generation larval body weights were disparate during development, while second generation larvae had similar weights with no significant differences. In this study we have tested the potential of captive breaded M. funereus larvae as a model for investigation of digestive enzymes. Amylase from the midgut of larvae reared under laboratory conditions showed twofold higher specific activities with a decreased number of isoforms expressed, as compared to the enzyme from field-collected larvae. Captive breeding of M. funereus can be used in the future as a part of an effective conservation strategy for this rare insect species.     

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.     

Life Cycle of the Mulberry Longicorn Beetle,Apriona germari Hope on an Artificial Diet

Mulberry longicorn beetle, Apriona germari, was reared on an artificial diet to investigate its life cycle. Average egg period was 17.95 days. Most larvae pupated at the 8th (33.3%) or 9th (33.3%) instars, but others emerged from the 7th to the 11th instars. The total duration of larval stage from the 1st to the end of the 9th and the 11th instars were 176.2 and 251.5 days, respectively. For terminatiopn of diapause and acceleration of pupation, low temperature treatment at 5°C for 60 days was highly effective. Average pupal period for female and male was 19.3 and 18.4 days, respectively. Average longevity of adults was similar with 40.5 days for female and 44.3 days for male. Mating had occurred from about 10 days after the adult emergence, and then a female adult laid one or two eggs per day. Average number of eggs oviposited by a female was approximately 47.7. Total life span of A. germari, when reared on an artificial diet in the laboratory, ranged broadly from 197.5 days to 331.5 days mainly depending on the larval period.     

Instar-specific defense ofColeomegilla maculata lengi (Col.: Coccinellidae): Influence on attack success of the intraguild predatorChrysoperla rufilabris (Neur.: Chrysopidae)

We conducted a laboratory study of the behavioural interactions between larval stages of the lady beetleColeomegilla maculata lengi Timberlake and an intraguild predator, the lacewingChrysoperla rufilabris Burmeister. Ladybird susceptibility to predation by third instarC. rufilabris is age-specific and influenced by larval defensive strategies. First and second instars ofC. maculata were captured more consistently by chrysopid larvae than third and fourth instars. Young coccinellids generally exhibit escape reactions (dropping, fleeing, retreating) but, do not survive after being caught by lacewings. In addition to escape reactions, older larvae may actively defend themselves by wriggling or biting, even after being pierced. The proportion of physical interactions that resulted in coccinellid death was significantly lower for older instars (<2%) than for first (35%) and second (19%) instars. Capture efficiency byC. rufilabris was related to the coccinellid-chrysopid size ratio. Dropping off the leaves, the most common defensive behaviour displayed by coccinellid larvae, contributed to reduce overall mortality of first instars to a level similar to the one observed for older stages. However, lacewings were able to prevent coccinellid dropping by impaling and holding their prey in the air, or by falling with the ladybirds to the ground where they continued their attack.     

Soybean leaf consumption by Nomuraea rileyi (Fungi: Deuteromycotina)-infected Plathypena scabra (Lepidoptera: Noctuidae) larvae

To determine the impact of Nomuraea rileyi on consumption by the green cloverworm, Plathypena scabra, larvae were reared from eggs obtained from field-collected moths, inoculated with conidia, and placed individually in separate plastic vials with a piece of surface-sterilized soybean leaflet. No significant differences in consumption rates were found between N. rileyi-inoculated and control larvae until after 144 hr post-treatment. After this period, consumption by larvae inoculated as first, second, third, or fourth instars was significantly less than that of control larvae because of mortality. First or second instars inoculated with N. rileyi conidia had significantly longer subsequent stadia than did control larvae. The average LT₅₀ of all inoculated instar groups was 6.5 days, and there was little evidence of significantly different N. rileyi susceptibility among instar groups. In general during the N. rileyi infection period leading to their deaths, inoculated P. scabra larvae had consumption patterns that were very similar to those of healthy larvae.     

Infectivity of an iridescent virus for larvae of Anticarsia gemmatalis (Lepidoptera: Noctuidae)

Laboratory tests were conducted with an iridescent virus (IV) of Anticarsia gemmatalis from Argentina to determine its infectivity for all six larval instars. For first, second, and third instars, the LC₅₀ values were 5.93, 6.14, and 11.30 mg/ml, respectively. The LD₅₀ values for fourth, fifth, and sixth instars were 1.21, 3.12, and 1.31μg/mg, respectively. The time until death was greater for early instars than for late instars; first instars averaged 23.1 days, second instars 19.6 days, third instars 19.5 days, fourth instars 14.3 days, fifth instars 8.2 days and sixth instars 6.5 days until death. High levels of iridescent virus inoculum appeared to activate a latent nuclear polyhedrosis virus in an average of less than 10% of larvae succumbing to a viral infection. Variable mortality rates were caused with low levels of IV inoculum; high dosages produced a high percentage of mortality, but it was virtually impossible to raise levels of inoculum to concentrations sufficient to produce 100% mortality.     

Development and feeding efficiency of Malacosoma neustrium larvae reared with Quercus spp. leaves

Observations on larval development of Malacosoma neustrium were conducted both in a cork oak stand and in the laboratory by using leaves of different host trees (cork oak, holm oak and downy oak) as food source. Instars were determined using head capsule and frass measurements. In the field the larvae progressed up to the fifth instar before pupating, and the increase in head capsule width followed Dyars Rule with a rate of increase (R.I.) value of 1.74. The same number of instars was determined for the larvae reared with cork oak (R.I. = 1.73) and holm oak (R.I. = 1.70) leaves. The caterpillars reared with downy oak foliage completed larval development in five, six and seven instars and the R.I. values obtained were 1.60, 1.52 and 1.44 respectively. A lower mortality was recorded for the larvae reared on holm oak. Growth and feeding indices were determined for the larvae from the third up to the last instar. The highest leaf consumption was detected for the fifth instar larvae reared on holm oak. For the caterpillars which completed five instars before pupating, the relative consumption rate (RCR) decreased from the third up to the fifth instar: from 4.8 to 1.7 (cork oak), from 7.4 to 3.3 (holm oak) and from 14.3 to 2.1 (downy oak). The relative growth rate (RGR) was highest during the fourth stadium (0.24, 0.27 and 0.33 for larvae reared with cork oak, holm oak and downy oak leaves respectively) and decreased in the fifth instar (0.09, 0.14 and 0.14 for larvae reared with cork oak, holm oak and downy oak leaves respectively), probably because of greater expense of energy due to the approach of maturity. Feeding and growth indices could be useful to define a defoliation prediction model.     

Prey selection and baculovirus dissemination by carabid predators of Lepidoptera

Abstract.

• 1 The interaction between coleopteran predators and baculovirus-infected larvae was studied in the laboratory and the field in order to assess the potential role of predators in the dissemination of a nucleopolyhedrovirus (NPV).
• 2 Preference tests using three carabid species, Harpalus rufipes De Geer, Pterostichus melanarius Illiger and Agonum dorsale Pont, showed no evidence of discrimination between healthy and diseased larvae of the cabbage moth Mamestra brassicae L. (Lepidoptera: Noctuidae) as prey items.
• 3 Virus infectivity was maintained after passage through the predator's gut. NPV mortality ranged from 97% to 20% when test larvae were exposed to faeces collected immediately after and 15 days post-infected meal respectively.
• 4 The potential for transfer of inoculum in the environment was estimated in the laboratory by soil bioassay. Carabids continuously passed infective virus to the soil for at least 15 days after feeding on infected larvae.
• 5 Field experiments showed that carabids which had previously fed on diseased larvae transferred sufficient virus to the soil to cause low levels of mortality in larval populations of the cabbage moth at different instars.

     

Observations onHexamermis albicans [Nematoda: Mermithidae] recovered fromLymantria dispar andStilpnotia salicis [Lep.: Lymantriidae] in west Germany and Austria

The mermithid,Hexamermis albicans (Siebold) was recovered from larvae ofLymantria (Porthetria) dispar (L.) collected from various localities in Burgenland, Austria in 1974 and 1975 and from Würzburg, Germany, in 1974. It was recovered also fromStilpnotia salicis (L.) in Austria in 1974. The mermithid was recovered from all field collected larval instars. The majority of the nematodes emerged fromL. dispar larvae collected as second and third larval instars although some nematodes were recorded from larvae collected as first instar larvae still on the egg mass. Peak emergence occurred in the laboratory during the period June 11–17 of both years, but emergence continued at a much lesser degree through the end of larval development. The nematode was found in both high and low host density populations. In 3 localities studied both years, there was a general increase in the percentage parasitism the second year. However, except for one locality in Austria in 1975 where individual samples produced up to 11% parasitism, the overall parasitism increased from 0.4% in 1974 to only 2.5% in 1975.     

The influence of understory nectar resources on parasitism of the spruce budworm Choristoneura fumiferana in the field

1 Nectar provided by flowering plants has been suggested as an important factor enhancing parasitism rates of herbivorous insects. Artificial nectar has been shown to increase parasitoid longevity and fecundity in laboratory studies. 2 We studied the influence of understory nectar on parasitism of the spruce budworm Choristoneura fumiferana in a field experiment in which we either removed understory flowering herbs or sprayed a sucrose solution on the understory vegetation in 0.25 ha quadrats. 3 Cohorts of laboratory-reared second instars were transferred the field to evaluate parasitism in the quadrats. The larvae were harvested as fifth instars and reared for parasitoids. 4 Parasitoids of early instars caused greater mortality in the supplemental sucrose treatment than in the vegetation-removal treatment or the control. Parasitoids of older larvae did not respond to the treatments. Larger-scale treatments may be necessary to elicit a response by the larger parasitoids that attack older larvae.     

The effect of gypsy moth metamorphosis on the development of nuclear polyhedrosis virus infection

The development of nuclear polyhedrosis virus (NPV) infection in gypsy moth (Lymantria dispar) was studied before, during, and after host metamorphosis, and in larvae and pupae in the subsequent generation, to determine whether NPV ingested by late instars can replicate in host tissues through metamorphosis and whether it can be vertically transmitted to progeny. Individuals that survived sublethal dosages of NPV did not differ from undosed insects in pupal weight, fecundity, larval and pupal weight of progeny, or response of progeny to NPV challenge. No evidence of NPV infection or of abnormal histology was found in adult tissues examined by light microscopy and no virus was detected on the surface of eggs produced by NPV-treated moths. No NPV-caused mortality was recorded among undosed progeny of dosed or undosed parents. The progeny of dosed parents were neither more resistant nor more susceptible to LdMNPV than were progeny of undosed parents and lethal times did not differ between groups. Examination of larval, pupal, and adult tissues by DNA hybridization revealed that insects in which NPV DNA was detected died prior to adult eclosion. NPV was not detected in any hosts surviving to the adult stage. These results suggest that survivors of sublethal dosages of NPV avoid infection and are therefore incapable of vertically transmitting infectious virus to progeny.