Virus pathogens of Culicidae (mosquitos).

II. VIRUS PATHOGENS OF CULICIDAE (MOSQUITOS) a Brian A. 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An iridescent virus with a particle size of 135-140 nm was isolated from larvae of Aedes stimulans collected from woodland pools in Connecticut. The virus appears to be a different strain than those isolated from other insect species and the name Aedes stimulans iridescent virus is proposed. Infected larvae were recognized by their opalescent turquoise colour. The incidence in the field was considerably less than 1%. The presence of the virus in ovarian tissue suggests transovarial passage. Anthony, D. W. & Hall, D. W. (1970). Electron microscope studies of the "R" and "T" strains of mosquito iridescent virus in Aedes taeniorhynchus (Wied.) larvae. Proc. 4th Int. Colloq. Insect Pathol., pp. 386-395. Electron microscope studies of the "R" (regular brown-orange iridescence) and "T" (turquoise iridescence) strains of mosquito iridescent virus in 4th-instar A. taeniorhynchus larvae showed the fat body to be the primary site of replication for both strains. In tissue sections, both strains appear hexagonal or pentagonal in shape, which suggests that each particle is an icosahedron. "T" strain is approximately 30-40 nm smaller than the "R" strain. Anthony, D. W. et al. (1973). A virus disease in Anopheles quadrimaculatus. J. Invertebr. Pathol., 22: 1-5. Free and occluded virus particles were found in the cytoplasm of the midgut epithelial cells of 2nd instar larvae of A. quadrimaculatus infected with Thelohania legeri. Spherical crystals, which contained these particles, showed a macromolecular para- crystalline lattice typical of polyhedral protein. In a few instances, the cuboidal crystals appeared to have coalesced to form larger crystals. The observations suggest that the free particles and their occluded forms may represent stages of a cytoplasmic polyhedrosis virus. Bertram, D. S. (1965). Double infection of mosquitoes with a virus and a malarial parasite. Proc. 12th Int. Cong. Entomol. (London), pp. 766-767. Malarial transmission by Aedes aegypti can be suppressed by concurrent infection of the vector with an arbovirus. The interaction may be fatal to the mosquito. Bird, R. G. et al. (1970). Evidence of insect viruses in colonies of Anopheles stephensi. Trans. Roy. Soc. Trop. Med. Hyg., 64(1): 28-29 (abstract). Three different virus-like particles were found in gut lesions of adult A. stephensi and a cell line derived from them. One was identified as a cytoplasmic polyhedrosis virus from the midgut and was associated with difficulty in malarial transmission. Another was found in the gut of individuals of the same strain but different colonies. The third type was observed in cell cultures and appeared to be TMV type rods. Chapman, H. C. (1972). Personal communication. 38 Chapman, H. C. (1974). Biological control of mosquito larvae. Ann. Rev. Entomol., 19: 33-59. Review article. Chapman, H. C. et al. (1970). Protozoans, nematodes, and viruses of anophelines. Misc. Publ. Entomol. Soc. Amer., 7: 134-139. Viruses such as the mosquito iridescent virus, cytoplasmic polyhedrosis viruses of the hypodermal and gut cells, and a nuclear polyhedrosis virus are reported only from culicines, but 2 probable cytoplasmic polyhedrosis viruses, one in the hypodermal cells and the other in the gut cells are reported from larvae of Anopheles crucians. Chapman, H. C. et al. (1969). A two-year survey of pathogens and parasites of Culicidae, Chaoboridae, and Ceratopogonidae in Louisiana. New Jersey Mosquito Extermin. Assoc. Proc., 56: 203-212. Adult and larval populations of mosquitos were sampled from 1967 to 1969. Forty-four species of mosquitos were captured and 35 were hosts to one or more pathogens. Inclusion and noninclusion viruses were recorded. Chapman, H. C. et al. (1966). Additional hosts of the mosquito iridescent virus. J. Invertebr. Pathol., 8: 545-546. Mosquito iridescent virus has been verified in Aedes taeniorhynchus, A. fulvus pallens, A. vexans, and Psorophora ferox from Louisiana. Infected larvae had an external iridescent hue that was usually first visible in the thorax of late 3rd- or early 4th- instar larvae. Most patently infected larvae die before pupation. Chapman, H. C. et al. (1972). Predators and pathogens for mosquito control. Amer. J. Trop. Med. Hyg., 21(5): 777-781. , The pathogens that can be or show promise of being manipulated and mass-reared in the laboratory are presented. Chapman, H. C. et al. (1967). Pathogens and parasites in Louisiana Culicidae and Chaoboridae. New Jersey Mosquito Exterm. Assoc. Proc., 54: 54-60. Twenty-seven species of mosquitos were found infected with internal parasites or pathogens in southwestern Louisiana. Mosquito iridescent virus was reported in some samples. Clark, T. B. (1972). Personal communication. Clark, T. B. & Chapman, H. C. (1969). A polyhedrosis in Culex salinarius of Louisiana. J. Invertebr. Pathol., 13: 312. A viral disease characterized by the presence of teragonal inclusion bodies in limb anlage and hypodermal cells was found in the larvae of Culex tarsalis in California. A similar virus has been found in C. salinarius in Louisiana. Not a true cytoplasmic polyhedrosis virus in that the inclusions studied in this paper are made up entirely of virus particles. This virus is most commonly referred to as a "tetragonal virus" because of the tetragonal nature of the inclusions. It is possibly a parvovirus. 39 Clark, T. B. et al. (1969). Nuclear polyhedrosis and cytoplasmic polyhedrosis virus infections in Louisiana mosquitoes. J. Invertebr. Pathol., 14: 284-286. A nuclear polyhedrosis virus infecting the gastric caeca and midgut of Aedes sollicitans and a cytoplasmic polyhedrosis virus infecting the gastric caeca and midgut of Culex salinarius were found in larvae collected in Louisiana. The most obvious sign of the disease was the whiteness of the gut wall due to the large number of inclusion bodies. Attempts at laboratory transmission of the nuclear polyhedrosis virus have yielded infection in only about 5% of the exposed larvae, but those infected died before pupation. The cytoplasmic polyhedrosis virus was easily transmitted in the laboratory but even heavily infected individuals pupated and emerged as apparently healthy adults. Clark, T. B. & Fukuda, T. (1971). Field and laboratory observations of two viral diseases in Aedes sollicitans (Walker) in southwestern Louisiana. Mosquito News, 31: 193-199. A viral epizootic involving both a cytoplasmic polyhedrosis and a nuclear polyhedrosis occurred in populations of Aedes sollicitans in Louisiana. Experimental results suggest that a series of overlapping broods of Aedes sollicitans may lead to a buildup of infective viral material in the habitat, but a period of drying between broods appears to reduce it very significantly. Transovum transmission and lateral transmission could explain the levels of infection found after the dry period. The introduction of infective material into an area previously almost free of disease resulted in a sig- nificant rise in the rate of infection. Clark, T. B. et al. (1965). A mosquito iridescent virus (MIV) from Aedes taeniorhynchus (Wiedemann). J. Invertebr. Pathol., 7: 519-521. Diseased 4th-instar larvae of A. taeniorhynchus were collected in Florida. Examination revealed a noninclusion virus with a dense central core surrounded by a 6-sided capsule which is composed of at least 2 layers. The principal site of infection is the cyto- plasm of fat cells. Infected larvae became iridescent orange during the 4th-instar and sometimes appeared milky white late in the disease. Per os transmission of MIV was achieved in only a small percentage of trials. Cunningham, J. C. & Tinsley, T. W. (1968). A serological comparison of some iridescent nonoccluded insect viruses. J. Gen. Virol., 3: 1-8. Tipula iridescent virus and Sericesthis iridescent virus were found to be serologically related by complement-fixation, tube-precipitation and agar-gel diffusion tests. They were unrelated to mosquito iridescent virus when compared by complement fixation. Dasgupta, B. (1968). A possible virus disease of the malaria parasite. Trans. Roy. Soc. Med. Hyg., 62: 730. Inclusion bodies have been observed in oocysts of Plasmodium. The abnormal oocysts showed partial to total loss of nuclei. Dasgupta, B. & Ray, H. N. (1957). The intranuclear inclusions in the mid-gut of the larva of Anopheles subpictus. Parasitology 47: 194-195. Feulgen-positive intranuclear inclusions were observed in the secretory cells of the mid- gut of larvae collected near Calcutta. A mature inclusion body appeared as a lump of DNA formed of globular bodies. Staining reactions suggested that the amount of DNA in the nucleus of other cells in the neighbourhood of the affected nucleus was reduced considerably. 40 Davies, E. E. et al. (1971). Microbial infections associated with plasmodial development in Anopheles stephensi. Ann. Trop. Med. Parasitol., 63: 403-408. Two forms of virus-like particles and a rickettsia-like organism have been observed in the midgut epithelial cells of a laboratory colony of A. stephensi. One of the viral particles does not appear to affect either the mosquito or the plasmodial infection. The other occurs in oocysts which show evidence of degenerative changes. Day, M. F. & Mercer, E. H. (1964). Properties of an iridescent virus from the beetle, Sericesthis purinosa. Aust. J. Biol. Sci., 17: 892-902. Sericesthis iridescent virus (SIV) was added to the medium in which second instar larvae of Aedes aegypti were developing. After two weeks the fat body in two living larvae became iridescent blue indicating infection by SIV. Faust, R. M. et al. (1968). Nucleic acids in the blue-green and orange mosquito iridescent viruses (MIV) isolated from larvae of Aedes taeniorhynchus. J. Invertebr. Pathol., 10: 160. Analysis of the nucleic acids revealed that DNA was present in both blue-green and orange MIV and that RNA was absent. DNA constituted 10.5% of the blue-green form and 11.7% of the orange. Federici, B. A. (1970). Unpublished observations. Federici, B. A. (1973). Preliminary studies on a cytoplasmic polyhedrosis virus of Aedes taeniorhynchus. Abstracts of papers, 5th Int. Collog. Insect Pathol. Microbial Cont. (Oxford), p. 34. The 4th-instar larva of A. taeniorhynchus from Louisiana was found infected with a cyto- plasmic polyhedrosis virus (CPV) in the posterior portion of the stomach. Infection trials gave an average rate of patent infection of 4.3% and mortality rate less than 1%. Occlusion bodies were present in the posterior stomach and occasionally in the gastric caeca. The coalescing process during occlusion formation is similar to that observed in a baculovirus in larvae of A. triseriatus. This similarity suggests that the mosquito host plays a role in the formation of occlusions. Federici, B. A. (1974). Virus pathogens of mosquitoes and their potential use in mosquito control. In: Aubin, A. et al., ed. Le controle des moustiques/Mosquito control. Quebec, Univ. Quebec Press, pp. 93-135. Review article with 56 references. Federici, B. A. & Anthony, D. W. (1972). Formation of virion-occluding proteinic spindles in a baculovirus of Aedes triseriatus. J. Invertebr. Pathol., 20: 129-138. An unusual process of inclusion formation was studied in A. triseriatus larvae infected with a Baculovirus (BV) similar to the nuclear polyhedrosis virus (NPV) type. In this disease virion-occluding proteinic inclusions initially developed individually. However, as the disease progressed the proteinic inclusions gradually coalesced eventually forming large rugose ellipsoids and finally, large smooth-surfaced spindles. Nuclei in late stages of infection usually contained two to five rugose ellipsoidal inclusions, frequently measuring 5 pm to 7 pm in diameter by 10 pm to 15 pm in length. The ellipsoidal forms exhibited different chemical behaviour from the spindles. 41 Federici, B. A. & Lowe, R. E. (1972). Studies on the pathology of a baculovirus in Aedes triseriatus. J. Invertebr. Pathol., 20: 14-21. The pathology of a Baculovirus (BV) in A. triseriatus was studied. The virus infected the cardia, gastric caeca, and the entire stomach of larval midgut epithelium. The progress of the disease was similar to that of other Baculoviruses of the nuclear poly- hedrosis virus (NPV) type. The disease differed from other BVs of the NPV type in that small proteinic inclusions gradually coalesced as they grew, forming large fusiform inclusions. Fukuda, T. (1971). Per os transmission of Chilo iridescent virus to mosquitoes. J. Invertebr. Pathol., 18: 152-153. Chilo iridescent virus (CIV) was successfully transmitted to 13 species of mosquitos, however the percentage contracting the disease was low. The CIV in mosquito larvae was first detected at the time of 3rd-instar when the blue-violet iridescence began to show. Death usually occurred in the 4th-instar. The virus had a wide host range in mosquitos and can be more easily mass-produced than MIV. Hall, D. W. & Anthony, D. W. (1971). Pathology of a mosquito iridescent virus (MIV) infecting Aedes taeniorhynchus. J. Invertebr. Pathol., 18: 61-69. RMIV was capable of infecting a variety of tissues within its host. Cells of the fat body, tracheal epithelium, imaginal discs, and epidermis were the primary sites of viral replication. Extensive destruction of the fat body by this virus resulted in the death of most infected mosquitos before they reached the adult stage. The transovarial trans- mission of RMIV was confirmed, and when transovarial transmission occurred, either all or none of the progeny of a given female were infected. Hall, D. W. & Fish, D. D. (1974). A Baculovirus from the mosquito Wyeomyia smithii. J. Invertebr. Pathol., 23: 383-388. A Baculovirus was found in larvae of W. smithii collected from a sphagnum bog in Massachusetts. This virus is similar in size and appearance to Baculoviruses from other mosquitos. A unique feature of the virus is the formation of polymorphic inclusions. The prepatent period for this virus is 3-5 days. Hall, D. W. & Lowe, R. E. (1971). A new distribution record for the mosquito iridescent virus (MIV). Mosquito News, 31: 448-449. Aedes taeniorhynchus larvae collected off the west coast of Florida were found to be infected with RMIV. About 0.12% of the specimens examined were infected. The location of these collections, in addition to those previously reported, suggests that RMIV is present throughout the range of A. taeniorhynchus. TMIV has not been found in Florida. Hall, D. W. & Lowe, R. E. (1972). Physical and serological comparisons of "R" and "T" strains of mosquito iridescent virus from Aedes taeniorhynchus. J. Invertebr. Pathol., 19: 317-324. Gel diffusion studies with alkaline degraded virus preparations exhibited four antigens common to both viruses but no unique antigens were detected for either virus. Electron micrographs of infected tissue sections showed tubular structures associated with both RMIV and TMIV. RMIV and TMIV sedimented at different rates in sucrose density gradients and RMIV was found to be slightly more dense than TMIV by equilibrium ultracentrifugation in cesium chloride. Mixtures of the two viruses can be separated by sucrose density gradient centrifugation. 42 Hasan, S. et al. (1970). Infection a virus irisant dans une population naturelle d'Aedes detritus Haliday en France. Ann. Zool. Ecol. anim., 2: 295-299 (English summary). An iridescent virus has been isolated from a naturally infected population of A. detritus in southern France. Morphologically and serologically the virus is identical to the iridescent virus described from this mosquito in Tunisia. (See Vago, Rioux, Duthoit & Dedet, 1969.) Hasan, S. et al. (1971). Infection of Aedes detritus Hal. with mosquito iridescent virus. Bull. World Health Organ., 45(2): 268-269. Attempts were made to infect larvae of A. detritus collected in France with MIV. Some larvae were kept for 24 hrs in water in which infected larvae had been macerated; others were infected with a virus suspension. In both cases, only 4th-instars showed disease symptoms, which appeared after 10 days for those injected and 20 days for those in the suspension. Infected larvae died within 2 weeks of appearance of symptoms. Hazard, E. I. (1972). Personal communication. Kellen, W. R. et al. (1963). A possible polyhedrosis in Culex tarsalis Coquillet (Diptera: Culicidae). J. Insect Pathol., 5: 98-103. Tetragonal intranuclear inclusion bodies have been observed in larvae collected from stagnant ponds in California. Infected larvae succumb in the 4th-instar, and success- ful transmissions of the disease agent have been obtained in the laboratory. It was concluded that an infectious agent is definitely involved, and that it might be a virus. Kellen, W. R. et al. (1966). A cytoplasmic-polyhedrosis virus of Culex tarsalis (Diptera: Culicidae). J. Invertebr. Pathol., 8: 390-394. The tetragonal crystals described by Kellen, Clark & Lindegren (1963) have been confirmed as inclusion bodies of a cytoplasmic-polyhedrosis virus* upon further examination. Polyhedra were present in hypodermal cells, in the developing leg, wing, and antennal buds. This is the first cytoplasmic polyhedrosis reported which is not restricted to gut tissue. Kelly, D. C. & Robertson, J. S. (1973). Icosahedral cytoplasmic deoxyriboviruses. J. Gen. Virol., 20(Suppl.): 17-41. A comprehensive catalogue of icosahedral cytoplasmic deoxyriboviruses (ICDV) isolated from animals and plants is given. The list includes 7 from mosquitos. Lebedeva, 0. P. et al. (1973). Investigation of a virus disease of the densonucleosis type in a laboratory culture of Aedes aegypti. Acta Virol., 17: 253-256. Histological changes in infected larvae were observed by light and electron microscopy. The most obvious pathological changes were in the cells of the fat body. The virus particles were about 200 A in size, having a paraspherical -shape and polygonal outlines. Observations revealed a similarity between the viral infection found in Aedes aegypti larvae and densonucleosis of Galleria mellonella. See footnote to Chapman et al., 1970. 43 Lebedeva, 0. P. & Zelenko, A. P. (1972). Virus-like formations in larvae of Aedes and Culex mosquitoes. Med. Parazitol. Parazit. Bolenzi., 41: 490-492 (Russian, with English summary). Inclusion bodies in the cytoplasm of fat body cells were found in 4th-instar larvae and pupae of Aedes aegypti and Culex pipiens molestus from laboratory colonies and in some Culex sp. from natural water bodies. The inclusions resembled in size, shape, and staining properties those of Entomopoxvirus described from other insects. Also, the diseased mosquitos exhibited the hypertrophied nuclei structurally similar to those described in densonucleosis of Galleria mellonella. (See Vago, 1963, and Vago et al., 1964.) Linley, J. R. & Nielsen, H. T. (1968a). Transmission of a mosquito iridescent virus in Aedes taeniorhynchus. I. Laboratory experiments. J. Invertebr. Pathol., 12: 7-16. The virus can be effectively transmitted to the larvae peros; transovarial transmission occurs from an infected female to her eggs, and evidence was obtained that all, or a very high proportion, of the eggs are infected. Larvae infected per os in their early instars may develop signs and symptoms of disease, and die before pupation. Exposure to infection progressively later in their development results in later development of signs and symptoms, and a reduction in the total number of larvae that show disease before pupation. Rearing larvae under different conditions of temperature, diet, and crowding produced no detectable differences in infection rates among either groups of larvae exposed from the time of hatching to the same dosage of virus, or groups of larvae hatched from a large batch of eggs in which a proportion carrying infection had been homogeneously distributed. Linley, J. R. & Nielsen, H. T. (1968b). Transmission of a mosquito iridescent virus in Aedes taeniorhynchus. II. Experiments related to transmission in nature. J. Invertebr. Pathol., 12: 17-24. A field experiment showed that infection could be acquired per os by larvae exposed under natural conditions, and that when healthy 4th-instar larvae were given access to intact diseased cadavers for a short time before pupation, they became infected and produced adults that laid infected eggs. Transovarial transmission of virus gives rise to diseased larvae, which die in the 4th-instar. The cadavers so formed provide the source of new infection, which is acquired per os by healthy larvae just before pupation. Infected adults from these larvae complete the cycle by depositing infected eggs. Lowe, R. E. et al. (1970). Comparison of the mosquito iridescent virus (MIV) with other iridescent viruses. Proc. 4th Int. Collog. Insect Pathol., pp. 163-170. The R isolate of the mosquito iridescent virus, although closely related to other insect iridescent viruses, possesses distinct physico-chemical characteristics. The diameter of RMIV (195 nm), molecular weight (2.486 x109 daltons), sedimentation coefficient (4.458), and DNA content (15.977%) are greater than the respective values for similar viruses. Although biologically similar, and showing no unique antigens serologically, the two colour isolates of MIV differ in size, sedimentation rate, and density. Matta, J. F. (1970). The characterization of a mosquito iridescent virus. II. Physico- chemical characterization. J. Invertebr. Pathol., 16: 157-164. A procedure for the purification of the R type of MIV is presented. The RMIV was distinctly different from the other iridescent viruses in terms of several physical parameters, and it would be difficult to justify considering it as a strain of these viruses. There was, however, a similarity in amino acid composition, indicating that they have a common phylogeny. 44 Matta, J. F. & Lowe, R. E. (1969). A differential staining technique for a mosquito iridescent virus. J. Invertebr. Pathol., 13: 457-458. A staining technique, that allows routine screening for MIV infections using a light microscope, was devised. Matta, J. F. & Lowe, R. E. (1970). The characterization of a mosquito iridescent virus (MIV). I. Biological characteristics, infectivity, and pathology. J. Invertebr. Pathol., 16: 38-41. Infected Aedes taeniorhynchus were mass-produced by rearing them in dishes containing homogenized infected 4th-instar larvae. The average mortality was dependent on crowding of the larvae during infection. The increase in virus over the inoculum varied between 70 and 310%. The RMIV infects only the fat body and imaginal discs in A. taeniorhynchus and destruction of these tissues is usually complete. Richardson, J. et al. (1974). Evidence of two inapparent nonoccluded viral infections of Culex tarsalis. J. Invertebr. Pathol., 23: 213-224. C. tarsalis from a laboratory colony in California were found to be infected with 2 noninclusion cytoplasmic viruses. Both viruses were seen in thin sectioned material from salivary glands, fat body, and nervous tissue of infected mosquitos. C. tarsalis virus 1 (CTV 1) was retained through 6 serial passages by needle inoculations of infected mosquito suspensions into virus-free adults. Both CTV1 and CTV2 appeared to multiply in the hosts. Infection may cause degeneration of salivary glands in adult females. Service, M. W. (1968). The ecology of the immature stages of Aedes detritus (Diptera: Culicidae). J. Appl. Ecol., 5: 613-630. Large numbers of larvae collected on Brownsea Island were infected by various pathogens including non-inclusion iridescent viruses. Stoltz, D. B. (1971). The structure of icosahedral cytoplasmic deoxyriboviruses. J. Ultrastruct. Res., 37: 219-239. The presence of two unit membranes in mosquito iridescent virus particles is clearly illustrated. The unit membranes in this study are defined on the basis of morphology alone. (See Stoltz, 1973.) Stoltz, D. B. (1973). The structure of icosahedral cytoplasmic deoxyriboviruses. II. An alternative model. J. Ultrastruct. Res., 43: 58-74. The available evidence seems to indicate that ICDV particles contain only a single structural unit membrane, associated with the viral nucleoid. Stoltz, D. B. et al. (1974). Virus-like particles in the mosquito Culex salinarius. J. Microscopie, 19: 109-112. Larvae of C. salinarius collected in Louisiana were found to have identical symptoms as those described in Culex tarsalis by Kellen, Clark & Lindegren (1966). The causative agent is readily transmissible to C. tarsalis. Although electron microscopic examina- tion revealed virus-like particles, the symptoms are not those expected from a typical cytoplasmic polyhedrosis virus. 45 Stoltz, D. B. & Summer, M. D. (1971). Pathway of infection of mosquito iridescent virus. I. Preliminary observations on the fate of ingested virus. J. Virology, 8: 900-909. MIV is ingested in large amounts by lst- and 2nd-instar Aedes taeniorhynchus larvae without causing a high rate of infection. Preliminary observations suggest that most, if not all, ingested particles are degraded shortly after entering the midgut. MIV and other virus particles were apparently unable to penetrate the peritrophic membrane; con- sequently none was observed inside, or in contact with, midgut epithelial cells. Tinsley, T. W. & Kelly, D. C. (1970). An interim nomenclature system for the iridescent group of insect viruses. J. Invertebr. Pathol., 16: 470-472. In the new system each iridescent virus is given a type number so additional host citing would pose no problem. This system is suggested to be employed until a system of more permanent names can be assigned based on comparisons of physical, chemical, and biologica properties of the various isolates. Tinsley, T. W. et al. (1971). An iridescent virus of Aedes cantans in Great Britain. J. Invertebr. Pathol., 18: 427-428. Larvae of A. cantans were collected from a pond in Kent, England, and found to be infected with a virus of the iridescent group. They developed a lime-green colour at an advanced stage of infection. Vago, C. et al. (1969). Infection spontanee a virus irisant dans une population d'Aedes detritus (Hal., 1883) des environs de Tunis. Ann. Parasit. Hum. Comp., 44: 667-676 (English summary). A virus disease was observed in natural populations of A. detritus in Tunisia. Diseased larvae could be identified by their slower movements and milky white coloration. Extensive damage to the fat bodies was observed. The purified virus has a cubical symmetry and measured 180 nm in diameter. It appears to be related to the iridescent viruses by its morphology, its location, and the iridescence of affected tissue. Wagner, G. W. et al. (1973). Biochemical and biophysical properties of two strains of mosquito iridescent virus. Virology, 52: 72-80. Several differing biophysical properties of the two strains (RMIV and TMIV) were observed due to size differences. The two strains appeared to be antigenically identical but the percent protein, DNA, and lipid of the two were dissimilar. Other than size variation, no morphological differences between RMIV and TMIV could be detected. Weiser, J. (1965). A new virus infection of mosquito larvae. Bull. World Health Organ., 33(2): 586-588. Diseased Aedes annulipes and A. cantans were found in South-western Bohemia. Infected individuals are opaque and opalescent, and are usually less motile. In their normal habitat, they appear to die before pupation. Nearly every tissue seems to be affected. Infection is not common, not more than 1% of the larvae are infected. In general appearance, the virus is similar to the Tipula iridescent virus. Wildy, P. (1971). Classification and nomenclature of viruses. Monogr. Virol., 5: 1-81. 46 Woodard, D. B. & Chapman, H. C. (1968). Laboratory studies with mosquito iridescent virus (MIV). J. Invertebr. Pathol., 11: 296-301. RMIV and a blue MIV of Aedes taeniorhynchus were serially passed through 68 and 30 generation, respectively, of larvae of A. taeniorhynchus. The average rate of infection for RMIV was 167% and 21% for the blue MIV. The maximum rate of infection was usually reached when large numbers of early-instar larvae were exposed for 48 hours to sub- stantial amounts of MIV material and the larvae were reared to the 4th stadium at 25°C. Attempts to transmit MIV to larvae of 9 species of mosquitos not known to be hosts were successful only with A. sollicitans. About 20%/ patent infection occurred in the larval progeny of adult A. taeniorhynchus derived from early 4th-instar larvae exposed to MIV. Such transovarial transmission was observed in the MIV of Psorophora ferox.