Genome of invertebrate iridescent virus type 3 (mosquito iridescent virus)

Iridoviruses (IVs) are classified into five genera: Iridovirus and Chloriridovirus, whose members infect invertebrates, and Ranavirus, Lymphocystivirus, and Megalocytivirus, whose members infect vertebrates. Until now, Chloriridovirus was the only IV genus for which a representative and complete genomic sequence was not available. Here, we report the genome sequence and comparative analysis of a field isolate of Invertebrate iridescent virus type 3 (IIV-3), also known as mosquito iridescent virus, currently the sole member of the genus Chloriridovirus. Approximately 20% of the 190-kbp IIV-3 genome was repetitive DNA, with DNA repeats localized in 15 apparently noncoding regions. Of the 126 predicted IIV-3 genes, 27 had homologues in all currently sequenced IVs, suggesting a genetic core for the family Iridoviridae. Fifty-two IIV-3 genes, including those encoding DNA topoisomerase II, NAD-dependent DNA ligase, SF1 helicase, IAP, and BRO protein, are present in IIV-6 (Chilo iridescent virus, prototype species of the genus Iridovirus) but not in vertebrate IVs, likely reflecting distinct evolutionary histories for vertebrate and invertebrate IVs and potentially indicative of genes that function in aspects of virus-invertebrate host interactions. Thirty-three IIV-3 genes lack homologues in other IVs. Most of these encode proteins of unknown function but also encode IIV3-053L, a protein with similarity to DNA-dependent RNA polymerase subunit 7; IIV3-044L, a putative serine/threonine protein kinase; and IIV3-080R, a protein with similarity to poxvirus MutT-like proteins. The absence of genes present in other IVs, including IIV-6; the lack of obvious colinearity with any sequenced IV; the low levels of amino acid identity of predicted proteins to IV homologues; and phylogenetic analyses of conserved proteins indicate that IIV-3 is distantly related to other IV genera.     

Field superparasitism by Diachasmimorpha longicaudata attacking Anastrepha spp. larvae on mango fruits

The incidence of superparasitism in field populations of Diachasmimorpha longicaudata (Ashmead), a solitary larval–pupal endoparasitoid, attacking Anastrepha fruit fly species was determined. From 14,550 fruit fly larvae infesting mango fruits in the field, 68.9% (10,038) survived to the pupal stage. Of these pupae, 3780 (37.7%) were parasitized, of which 2075 (54.9%) were superparasitized. The number of oviposition scars per pupa ranged between 1 and 29, although high numbers were rare. The presence of one or two scars per host was frequent, but only 8.6% of pupae had more than three scars. Fruit size was positively related to the level of infestation by Anastrepha spp. and was significantly correlated with the numbers of parasitized and superparasitized pupae by D. longicaudata. The length and width of host pupae were positively related to superparasitism, and superparasitism was positively correlated with a sex ratio biased toward females. Our data show that superparasitism is present in natural populations of D. longicaudata at remarkable rates and confers some advantageous features such as a female-biased sex ratio. These findings favor the election of this species as a viable biocontrol agent for augmentative releases because the favorable proportion of adult females emerging from superparasitized pupa should contribute to better pest control.     

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.     

Resistance of cocoa to swollen-shoot virus in Ghana. I. Field trials

In Ghana, field resistance to cocoa swollen-shoot virus (CSSV) in young, bearing trees was assessed by monitoring natural spread by mealybug vectors from adjacent graft-inoculated line sources. Apparent rates of virus spread were estimated by regressing proportions of infected trees on time from inoculation. In some hybrids between parents derived from Upper Amazonian material, spread was only 25–40% that in currently recommended varieties. This resistance is the most effective, feasible measure to reduce economic losses and the resistant hybrids could be made available quickly for large scale use. The results from trials in which virus spread most rapidly were the easiest to interpret. Spread of CSSV is erratic, necessitating large experiments, and in trials with equal replication the largest plots gave the greatest precision. Neighbouring plot adjustment proved unsatisfactory as it improved precision but appeared to reduce accuracy.     

Field trials on transmissible gastroenteritis live virus vaccine in newborn piglets.

Field trials were conducted on attenuated live virus vaccine of transmissible gastroenteritis to confer active immunity to newborn piglets. To examine innocuity and efficacy of the vaccine, a total of 714 newborn piglets were subjected to these trials. Of them, 357 piglets were administered orally with 10(7.0) TCID50 within 3 days after birth, and the other 357 piglets served as nonvaccinated controls. No undesirable postvaccinal reaction was observed in any vaccinated piglet. Suckling piglets born from nonimmune sows showed a good antibody response after vaccination. They were different, however, in antibody titer from one experimental place to another. Antibody levels were high in piglets raised in the northern experimental places. On the contrary, the antibody response of suckling piglets born from immune sows was influenced by vaccination. In most of these piglets, antibody titers declined markedly and disappeared finally 3 months after vaccination. About 25% of the non-vaccinated piglets showed an antibody response by pen contact with vaccinated ones.     

Species specific responses of common grassland plants to a generalist root herbivore (Agriotes spp. larvae)

Insect root herbivores can alter plant community structure by affecting the competitive ability of single plants. Our study aims at increasing knowledge on the impact of click beetle larvae (Elateridae, genus Agriotes) on grassland plant communities, by determining biomass responses as well as responses of the arbuscular mycorrhizal fungi (AMF) symbiosis to this widespread generalist root herbivore for eight common grassland plant species (Festuca rubra, Holcus lanatus, Poa pratensis, Achillea millefolium, Plantago lanceolata, Veronica arvensis, Medicago lupulina, Trifolium repens), belonging to three functional groups (grasses, herbs, legumes).The presence of larvae in the rhizosphere of individual plants had an overall negative effect on root biomass, which varied in degree between plant species, with more root biomass being removed from larger root systems. The effect of larvae on shoot biomass, total plant biomass and shoot/root ratio also differed in strength between plant species. Relative changes in root and total plant biomass ranged from a 71% and 55% loss, respectively, in V. arvensis to an 11% and 1% increase in T. repens, but were not related to root- or plant size. Root colonization by AMF and the length of extraradical AMF hyphae were not affected by larvae. The plant's functional group did not determine the response of the plant to feeding by larvae. Growth of larvae was positively correlated with root biomass, but did not depend on plant species or group identity.The results confirm the generalist nature of Agriotes spp. larvae, which depend on sufficient root quantity and are likely to feed most on those plant species whose roots are most abundant in their habitat. Their effect on the plant community may be generated through the ability of the respective plant species to cope with the herbivore damage, with tolerances being plant species rather than plant group specific.     

Parasitoid-mediated transmission of an iridescent virus

We examined the interaction between an invertebrate iridescent virus (IIV) isolated from Spodoptera frugiperda (J.E. Smith) and the solitary ichneumonid endoparasitoid Eiphosoma vitticolle Cresson. In choice tests, parasitoids examined and stung significantly more virus infected than healthy larvae, apparently due to a lack of defense reaction in virus infected hosts. Parasitoid-mediated virus transmission was observed in 100% of the female parasitoids that stung a virus infected host in the laboratory. Each female parasitoid transmitted the virus to an average (+/-SE) of 3.7+/-0.3 larvae immediately after stinging an infected larva. Caged field experiments supported this result; virus transmission to healthy larvae only occurred in cages containing infected hosts (as inoculum) and parasitoids (as vectors). The virus was highly detrimental to parasitoid development because of premature host death and lethal infection of the developing endoparasitoid. Female parasitoids that emerged from virus infected hosts did not transmit the virus to healthy hosts. We suggest that the polyphagous habits of many noctuid parasitoids combined with the catholic host range of most IIVs may represent a mechanism for the transmission of IIVs between different host species in the field.     

The biology of Chilo iridescent virus

Chilo iridescent virus (CIV) is the type species for genus Iridovirus, and belongs to the family Iridoviridae. Since the discovery of CIV in 1966, many attempts were made to elucidate the viral genome structure. The virions contain a single linear ds DNA molecule that is circularly permuted and terminally redundant. The genome of CIV has been entirely sequenced. The CIV virion consists of an unusual three-layer structure containing an outer proteinaceous capsid, an intermediate lipid membrane, and a core DNA-protein complex containing the genome. CIV has a broad host spectrum and has, in general, a limited mortality effect on its hosts. Up to now there have been several studies about CIV describing its structure, ecology, and molecular biology. In this review study we present all these studies together to describe the CIV.     

Persistence of Invertebrate iridescent virus 6 in soil

Soil represents an important reservoir for mostentomopathogenic viruses. Invertebrateiridescent viruses (IIVs) (Iridoviridae) arenon-occluded DNA viruses that infectagriculturally and medically important insectspecies, especially in damp or aquatichabitats. We used virus extraction and insectbioassay techniques to determine the effect ofsoil moisture and soil sterility on thepersistence of Invertebrate iridescentvirus 6 (IIV-6) in a soil over a 90 day periodin the laboratory. Loss of activity of IIV-6in dry soil (6.4% moisture, –1000 kPa matricpotential) was very rapid and was not studiedbeyond 24 h. Soil moisture did not affect therate of inactivation of virus in damp (17%moisture, –114 kPa matric potential) or wetsoil (37% moisture, –9.0 kPa matricpotential). In contrast, soil sterilizationsignificantly improved the persistence of IIV-6activity, both in damp and wet soil. Controlvirus suspensions retained 0.72–0.87% oforiginal activity after 90 days, which wassignificantly more than the activity retainedin soil. These figures represent half lives of4.9 days for IIV-6 in non-sterile soil, 6.3days in sterilized soil (data pooled formoisture treatments), and 12.9 days for thecontrol virus suspension. We conclude thatextra-host persistence in soil habitats may bean important aspect of the ecology of IIVs.     

An iridescent virus fromPopillia japonica (Col.: Scarabaeidae)

A very low incidence (<0.01%) of a blue iridovirus (IV) was found in larvae of the Japanese beetle,Popillia japonica Newman, that were sampled over a two year period on Terceira Island (Azores, Portugal). In the most heavily infected larvae, a deep blue iridescence was observed, particularly in the fat body. Transmission electron microscopy revealed the characteristic crystalline arrays of the hexagonal virus particles in the cytoplasm of fat body cells, tracheal matrix, muscle, hypodermis and blood cells. Crystals of the virus particles were also observed freely circulating in the hemolymph. The average diameter of negatively stained purified virus particles was 157 nm. Similarities and differences with other IVs found in the Scarabaeidae are discussed. Considering the broad host range of some of the iridescent viruses, the relatively recent invasion of Terceira byP. japonica, and the rarity of the virus in the beetle, it is probable that the infection was the result of transmission from another species of soil-inhabiting arthropod. Its value as a potential biological control agent ofP. japonica is negligible.     

Spatial distribution of bibionid larvae in agricultural grassland

The distribution of bibionid larvae in agricultural grassland was investigated. They tend to occur sporadically and at low population density. More detailed sampling within a field showed that numbers were highest adjacent to a hedgerow with most being found 6–11 m from the hedge. Log variance/log mean relationships were established for counts of Bibio johannis and Dilophus febrilis larvae collected from soil cores in grassland. Both species were highly aggregated. An area of grassland (160 cm × 80 cm) was divided into 128 soil blocks and B. johannis larvae extracted. Almost all larvae were associated with aggregations. The two-phase mozaic method of Pielou (1974) was applied to larval counts from different sized concentrically collected soil cores. Larval patches were not randomly distributed. Time-lapse video studies of bibionid larvae were made and showed both aggregation and processional behaviour. This was associated with chemical stimulation which is probably used to maintain larval groupings in the field. There was evidence for two levels of aggregation. The first is due to groups of larvae arising from eggs laid by a single female and the second, larger scale, aggregation is due to factors that influence the distribution of adult flies.     

Field trials of CpGV virus isolates overcoming resistance to CpGV-M

The Cydia pomonella granulovirus (CpGV) has been used for many years as biological agent for codling moth control in apple orchards. Resistance to the Mexican strain of CpGV was detected in orchards in Germany, France and Italy. A laboratory insect colony was started from insects collected in a French resistant orchard. It was named RGV. Various virus isolates were identified as active against this resistant insect colony. Field tests were carried out in 2007 to test if the two virus isolates CpGV-I12 and NPP-R1 were effective in the field. Although these virus isolates were not able to reduce insect caused fruit damages, they significantly reduced the overwintering insect populations. NPP-R1 was subjected to eight passages on RGV larvae (NPP-R1.8) that improved its biological activity on RGV larvae. 2008 field trials were set up to test this improved virus strain, compared to CpGV-I12 and Madex plus active on RGV. These tests confirmed the ability to control both in susceptible and resistant insect populations.     

Multiplication of Chilo iridescent virus in noninsect arthropods

Attempts to infect noninsect arthropods with Chilo iridescent virus (CIV) originally isolated from Lepidoptera were made by using eight species belonging to four classes. Multiplication of CIV was demonstrated in two species of terrestrial Crustacea (the pill bug, Armadillidium vulgare, and the slater, Porcello scaber) and one species of Chilopoda, the house centipede, Thereuonema higendorfi. The lethality experiment of CIV for A. vulgare suggested that chronic infection is a characteristic of the CIV infection in both classes, Crustacea and Insecta. Neither iridescence nor recovery of virus infectivity was demonstrated in the following arthropod species: the sea slater, Ligia exotica (Crustacea: Isopoda), the grapsid crab, Sesarma haematocheir (Crustacea: Decapoda), the millipede, Oxidus gracillis (Diplopoda: Polydesmoidea), Rhysodesmus semicirculatus (Diplopoda: Polydesmoidea), and the giant crab spider, Heteropoda venatoria (Arachnida: Araneae).