Repeatedly granulosis viruses have been observed to be responsible for severe epizootics in various insect populations. An often recurring example is that displayed by the worldwide spread granuloses of Pieris spp.
As Todd (1960) points out, a series of field surveys during the 1955-1958 seasons in the North Island of New Zealand revealed that Pieris rapae "is effectively checked each season by a virus disease." From the symptoms described, the causative agent was a granulosis virus. No doubt, it was the same virus that "exercised an unusual high degree of control" of the same pest in 1947-1948 in New Zealand (Kelsey, 1957).
In 1957 in that country the average natural granulosis of P. rapae rose to about 77 percent in March, and in several cases even complete control of the larvae was recorded (Kelsey, 1958). Also in Hawaii the same granulosis virus was observed to cause "severe epidemics, which often result in the control of the cabbageworm" (Tanada, 1956c). In Japan granulosis apparently contributes to the reduction of populations of Pieris rapae crucivora Boisduval (Ito et aL, 1960). Smith and Rivers (1956) supposed that the granulosis of Pieris spp. was introduced into England by a very heavy invasion of white butterflies from the continent in 1955. Indeed, in the European continental countries Pieris granuloses are very common. In 1951 Thompson reported a granulosis in an insectary stock of P. rapae in California. So far, this disease has not been found in the field there. In recent infection experiments by Martig
noni and Schmid (1961) larvae of two California populations of P. rapae showed no differences in susceptibility to granulosis.
564 ALOIS HUGER
An impressive example for the effective destruction of a population of Eucosma griseana within a period of two years by a granulosis epi
zootic is reported by Martignoni (1957). At the culmination point of the gradation, up to about 80 percent of the larval populations were destroyed by granulosis (Auer, 1956, cited in Baltensweiler, 1958).
According to Martignoni (1957), the L D5 0 of the granulosis virus in
creased during the breakdown of the population from 1954 to 1955, probably as a result of selection (see Section V I I I , C ) . It seems that stress conditions involved in a complexity of factors triggered this epi
zootic wave. T h e outbreak of the epizootic is in causal relation with the high population density (as associated with the "carrying capacity" of a certain biotope) and with the scarcity of food (starvation) (Martignoni,
1957; Baltensweiler, 1958).
There are other factors inducing epizootics or effective natural limitation by granulosis. It is suggested by Tanada (1956b, 1959b, 1961) that in the case of Pseudaletia unipuncta synergism between a granulosis virus and a nuclear polyhedrosis virus may play an important part in the development of virus epizootics in the field (see Section V I I I , C, and D ) . Epizootiological studies from 1958 to 1960 in Hawaii indicated the two viruses to be among the important natural factors regulating the population of P. unipuncta during the spring months (Tanada, 1961).
Kova£evic (1958) underlines that in the latently infected populations of Hyphantria cunea unfavorable conditions of weather and food induce granulosis epizootics and bacterioses, especially in the second generation, thus regulating the population density (see Section V I I I , F ) . For the realization of the latter he concluded that latent infections are more important than those acquired perorally and otherwise.
T h e action of granulosis as a natural mortality factor in insect populations must not necessarily depend on high density. Hall (1953), for example, stated that Autographa californica was effectively controlled under conditions of very low host density by granulosis, polyhedrosis, and certain insect enemies. Thus the level of population density necessary for effective natural limitation differs widely, although high population density, in general, offers better chances for the development of epizo
otics.
Other granuloses which may cause high mortality in the field, even of epizootic proportions, have been observed in populations such as those of Choristoneura murinana (Bucher, 1953; Capek et al., 1958;
Franz, unpublished); Harrisina brillians (Hall, 1955; Smith et al., 1956; Clausen, 1958, 1961); Amelia pallorana, Megalopyge opercularis (Steinhaus, 1957); Eulype hastata (Linnaeus) (Steinhaus, 1959); Hyphan
tria cunea (Kovacevic, 1958; Schmidt, 1959); Pseudaletia unipuncta
(Ta-nada, 1956b, 1959b, 1961; Raun, 1961); Persectania ewingii (Lower, 1954).
Although in populations of Recurvaria milleri mortality due to granulosis ranged between 30 and 50 percent in 1953, in the following years a heavy increase of the pest took place, and the mortality caused by granulosis remained at an enzootic level (Struble and Martignoni, 1959). T h e latter seems often to be the case with granuloses and, in general, applies to that of Argyrotaenia velutinana, first discovered by Sibold in 1950, and the virus described by Wasser and Steinhaus in 1951, and the disease studied in the field by Schoene and Sibold (1951, 1952), Glass (1958), and Oatmen and Jenkins (1962).
T h e spread of granulosis infections in nature is achieved in various ways. T h e transmission of the granulosis virus to individuals of the same generation (horizontal transmission) mainly occurs by ingestion of contaminated food. Contact with infected specimens and cannibalism further may help to spread granulosis (Lower, 1954). Especially from larvae of Pieris spp. it is known that not infrequently they contract the disease by feeding on liquefied cadavers of virus-killed larvae which seem to have an attraction for them (Smith, 1959a, b, 1960; Wilson, 1960).
Rain may splash the capsules liberated from the cadavers all over the plant or transfer them from the soil on to lower leaves of plants, such as cabbage, by splashing around (Kelsey, 1958, 1960; Todd, 1960), and wind may carry them still further afield. Arthropods, especially preda
cious ones, and birds are potential vectors of the capsules (Franz, 1956;
Smith et al, 1956; Kelsey, 1960; Todd, 1960; Clausen, 1961).
Parasitic insects repeatedly have been indicated as being instrumental in the mechanical transmission of the granulosis viruses by their con
taminated ovipositors (Lower, 1954; Smith et al, 1956; Kelsey, 1957;
Todd, 1960; Wilson, 1960). Indeed, Kelsey (1960) succeeded in demon
strating in the laboratory that seven of ten Apanteles glomeratus (Lin
naeus) that had oviposited in infected larvae of P. rapae were able to transmit the granulosis virus to healthy Pieris larvae. However, it is not proved whether this method of virus transmission really occurs naturally in the field. At any rate, no Apanteles have been observed ovipositing in infected, discolored Pieris larvae. On the other hand, Smith et al (1956) observed an apparent correlation between natural outbreaks of the granulosis of Harrisina brillians and the presence of the two major insect parasites Apanteles harrisinae Muesebeck and Sturmia harrisinae Coquillett. T h e i r laboratory experiments led to the same conclusion.
Parasitic insects developing in virus-infected hosts are not directly affected by the virus, nor do virus-infected host larvae always die before the parasites mature and emerge (Lower, 1954; Tanada, 1956b; Wittig,
566 ALOIS HUGER
1959a, b; Kelsey, 1960; Wilson, 1960). Often, however, the host larvae succumb to granulosis before the parasitic larvae are fully fed; then the latter die, presumably of starvation, thus often being greatly reduced in number in the parasite population (Smith et al., 1956; Baltensweiler, 1958; Kelsey, 1960; Wilson, 1960; Clausen, 1961).
Another method of dissemination of the granulosis viruses, as with other insect viruses in an active or latent state, is by trans-ovum trans
mission from parent to offspring. Though it is very difficult or, for the time being, impossible to obtain clear-cut evidence of this so-called vertical transmission by exact demonstration of capsules, virus rods, or constituents of them in or on the eggs, there are many observations which strongly support the conclusion of the trans-ovum or transovarian passage of the granulosis viruses (e.g., Steinhaus, 1947; Biliotti et al., 1956;
Smith et al, 1956; Martignoni, 1957; Bergold, 1958; Rivers, 1959; Sager, 1960; Smith, 1960). Smith et al. (1956) even succeeded in demonstrating capsules in the embryos of H. brillians.
No doubt, this trans-ovum passage of insect viruses in many cases is of primary significance for their dissemination and thus for the outbreak of epizootics.
For the longevity of granulosis viruses see Section V I I . B. Biological Control
By "biological control" here is meant the artificial application of granulosis viruses in the field for the control of insect pests. According to the quite different experimental conditions and circumstances, experi
ments in this direction have led to different results. Martignoni and Auer (1957) conducted a pilot field test in the control of Eucosma griseana by spraying 5 liters per tree of a capsule suspension containing 18 χ 1 07 capsules per milliliter, but without success. This failure probably can be explained by the fact that the experiment was carried out in a declining population suffering from a granulosis epizootic where the L D5 0 had been increased (Martignoni, 1957; see Section V I I I , C ) . This test at least demonstrated the necessity of clarifying important points (such as the dosage of virus, the susceptibility of the population, and the timing of the application in relation to the larval age and phase of gradation) before large scale experiments are initiated. In addition, the findings of Sidor (1959) that larvae of Pieris brassicae from different localities varied in their susceptibility to their own granulosis and cytoplasmic polyhedrosis virus underline this point (see Section V I I I , C ) .
Glass (1958, 1959) conducted field tests to control Argyrotaenia velutinana by spraying a suspension of triturated diseased larvae at rates of 5, 50, and 100 per gallon to apple trees during the hatching of the first
generation. All three concentrations proved highly virulent and ulti
mately lethal. Most of the larvae contracted granulosis, but they were not inactivated before they had damaged many fruits.
Investigations by Glass (1958) on the effect of certain fungicides, at the concentrations normally used in the field, on the granulosis viruses of A. velutinana showed that elemental sulfur, ferbam, captan, dichlone, and phenylmercury acetate ("Tag") did not reduce the incidence of the disease, whereas glyodin did.
T h e most successful experiments in the biological control of pests by granulosis viruses have been carried out so far with populations of Pieris rapae in Hawaii (Tanada, 1953c, 1956c, d ) , and New Zealand
(Kelsey, 1957, 1958, 1960; Wilson, 1960), and with P. brassicae in France (Biliotti et aL, 1956). Small-scale tests with both host species are reported from Great Britain (Smith, 1959b, 1960). T h e moderate success of field tests with the granulosis virus of P. rapae at Geneva, New York, by McEwen and Hervey (1959) apparently is due to the particular experi
mental conditions involved.
In the above-mentioned field tests by Tanada (1956c, d) effective control was obtained by application of sprays prepared by suspending in a gallon of water only two fifth-instar larvae of P. rapae that had succumbed to granulosis. Similarly, Smith (1959b, 1960) found that five mature larvae of Pieris spp. in an advanced stage of, or killed by, granulosis are sufficient to prepare one gallon of highly infectious spray.
As to the production of larger quantities of capsules for wide-scale use in biological control, so far there is only the possibility of multiplying or producing the viruses by infection of the hosts, if enough cadavers of virus-killed specimens cannot be collected directly from the field. Limited attempts to propagate two granulosis viruses in chick embryos were unsuc
cessful (Steinhaus, 1951). Progress in insect tissue culture possibly will open new ways.
According to Smith (1960), it is no problem to build up large sup
plies of the granulosis virus of P. brassicae, since this host can be mass reared all the year round (David, 1957). For a somewhat different method of multiplying the same virus, published by Vago and Atger
(1961), see Section V I I I , C.
T h e effect of storage and temperature on the infectivity of the granulosis viruses is treated in Section V I I .
X . T A X O N O M Y OF GRANULOSIS VIRUSES
When Holmes (1948) made an attempt to classify insect viruses, applying Linnaean binominals, he included in the genus Borrelina members "known only as attacking lepidopterous insects." This genus
568 ALOIS HUGER
comprised polyhedral viruses as well as viruses causing granuloses. In view of the fundamental differences of these two types of insect viruses, Steinhaus (1949a, b) created the generic name Bergoldia for the granu
losis viruses, with the type species Bergoldia calypta Steinhaus. In further contributions on the taxonomy of insect viruses (Bergold, 1953c; Stein
haus, 1953), the leading criteria proposed by the Virus Subcommittee of the International Committee on Bacteriological Nomenclature for the description of genera and species (Andrewes, 1951) have been accepted for insect viruses, too. Zhdanov (1957), disregarding existing names, suggested the generic name "Capsulatus" for granulosis viruses, yet this is to be regarded as invalid because of the priority of the generic name
"Bergoldia." Zhdanov himself later rescinded his proposed names (Ber
gold, 1959b). Another proposal, in 1953, made by the Committee men
tioned above was "Capsula-virus" but without authors (Andrewes, 1954, 1955). However, the Insect Virus Study Group (belonging to this Com
mittee) finally accepted the "-virus" suffix at the end of the original ge
neric name Bergoldia, but in conjunction with the species name as well as the author's name (Bergold et al., 1960). T h e type species, therefore, is called Bergoldiavirus calyptum Steinhaus.2 Accordingly, in T a b l e I the existing names of granulosis viruses have been combined in this sense.
Included herein is the change of the gender endings of specific names to agree with the gender of the generic name according to the rules of the International Code of Nomenclature of Bacteria and Viruses.
Referring to the reported differences in the site of development of capsules in the host cells (see Section V I I I , A ) , Weiser (1958) proposed two genera: (1) cytoplasmic granuloses with the genus name Bergoldia Steinhaus 1949, and (2) nuclear granuloses with the new genus name of Steinhausia. T h e contradictory cytopathological findings obtained so far (see Section V I I I , Β , 1 and 2) show that further careful investigations are necessary before these distinctions can be possibly applied to granu
losis viruses. Nevertheless, such nomenclatorial designations may be valid if true nuclear and true cytoplasmic granuloses are eventually verified.
ACKNOWLEDGMENTS
T h e author wishes to thank Dr. J . M. Franz and Dr. A. Krieg from our laboratory for critical reading of the manuscript. He is further indebted to Dr. G. H. Bergold, Dr. G. H. Hills, Dr. Κ. M. Smith, Dr. N. Xeros, and Dr. Y. Tanada for photographs.
2 Some students of virus nomenclature feel that this (and other similar names of insect viruses) is not really a new combination, since a new combination usually indicates the transfer of a species to another genus. In the present case no new genus or such transfer is involved, there is only a modification in the spelling of a pre
viously published generic name, in other words, an emendation.—Editor.
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