Paper 1.8

Am. J. Trop. Med. Hyg.. 34(2),1985,pp. 406-412 Copyright @ 1985by The American Societyof Tropical Medicineand Hygiene

THE ISOLATION OF ARBOVIRUSES INCLUDING A NEW FLA VIVIRUS AND A NEW BUNY A VIRUS FROM (CERA TIXODES) URIAE (IXODOIDEA: ) COLLECTED AT MACQUARIE ISLAND, , 1975-1979*

T. D. ST. GEORGE,' R. L. DOHERTY,2 J. G. CARLEY,2C. FILIPPICH,2 A. BRESCIA,3J. CASALS! D. H. KEMP,. ANDN. BROTHERS' 'CSIRO. Division of Tropical Science.Indooroopilly. Brisbane4068, Queensland.Australia. 2QueenslandInstitute ofMedical Research. Bramston Terrace.Brisbane 4006. Queensland.Australia. 3Yale ArbovirusResearch Unit. Yale UniversitySchool ofMedicine. New Haven,Connecticut. USA(present address: Department ofNeurology. The .'vfountSinai School ofMedicine. New York. New York. USA). and .Department ofNational Parks and Wildlife. Hobart, Tasmania,Australia

Abstract. Pools of , Ixodes (Ceratixodes)uriae collected between 1975 and 1979 at Macquarie Island, yielded 33 strains of at least 4 different viruses: Nugget virus (Ke- merovo group), 1 strain; Taggertvirus (Sakhalingroup) 9 strains; a previously undescribed flavivirus, related to Central EuropeanTickbome encephalitis virus, for which the name "Gadgets Gully" is proposed, 9 strains; a virus serologicallyrelated to the Uukuniemi serogroup,family Bunyaviridae,** for which the name "Precarious Point" is proposed, 10 strains. Three isolates were mixtures of Nugget and GadgetsGully viruses; the remaining virus strain remains unidentified.

Macquarie Island, situated in the southernPa- are associated with the penguin colonies. The cific Ocean (latitude 54°30'S,longitude 159°E), viruses Nugget (Kemerovo Group) and Taggert is a national park with its only human inhabit- (Sakhalin Group) were isolated by Doherty et al. I ants a small party of scientists. At sea level the from ticks of this species collected on Macquarie air temperaturevaries between0 and 10°Cfrom Island. This report describes a continuation of winter to summer. Precipitation of 2,000 mm is that study. spread throughout the year with some rain al- most every day. Seabirds are abundant. Large populations of ticks Ixodes (Ceratixodes)uriae MATERIALS AND METHODS Macquarie Island (Fig. I) is 30 km long by 5 km at its widest and has rocky, narrow fore- Accepted 12 September1984. shores,fringing cliffs on the eastcoast, and many * Arbovirus researchat the QueenslandInstitute of pebbly beaches between headlands. The west coasthas a narrow coastal plain covered with a Medical Research (QIMR) was supported by the QueenslandState Government and Researchat the densemass of matted vegetation.There is a cen- Yale Arbovirus ResearchUnit (Y AR U) wassupported tral plateauof 200-420 meters. It is on the beach by NIH Grant AI 10984 Contracts DAMD-17-8l-C- areas and slopes immediately inland that the 1121from theU.S. Army and NOOOI4-78-C-Ol04 from penguincolonies are found. All of the collections the U.S. Navy, and grants from CSIROand the World Health Organization. of the I. (C.) uriae were made from under Address reprint requeststo: Librarian, CSIRO, Di- rocks and debris or in tussockgrass (Poafoliosa) vision of Tropical Animal Science,Long PocketLab- in the vicinity of these colonies (Table 1). No oratories, P.B. No.3, P.O., Indooroopilly, Brisbane, collectionswere made from plateau or cliff areas Queensland4068, Australia. ** PrecariousPoint virus was recentlyfound to be a where other seabirds nest. Ten widely spaced member of the Uukuniemi serogroup,Bunyaviridae at sitesnear the seashorewere sampled. The species YARU. of penguiw inhabiting rookeries on Macquarie

4( )6 F'aper1.8

NEW ARBOVIRUSES AT MACQUARIE ISLAND 407

oculated into baby hamster kidney monolayers (BHK2l).3 The cells were grown in Hanks' bal- ancedsalt solution with 0.5% lactalbumin 0.01% Difco yeast extract, 10% fetal calf serum, and 1,000units eachof sodium penicillin and strep- tomycin sulphate per mI. The medium was re- moved the day after inoculation with tick ma- terial and the cell sheetwas washed to remove visible fragments of ticks. The medium was re- placedwith the growth medium in which the fetal calf serum content was reduced to 4%, and 50 units/ml of Nystatin were added. The cell sheets wereobserved 3 times a week for cytopathology. If cytopathologywas noticed, the cells were pas- sagedto further BHK2l monolayers. All mono- layerswithout cytopathologywere passaged twice at weeklyintervals beforebeing discardedas neg- ative. Any virus isolated in cell cultures was adapted by intracerebral injection of mice for preparation of mouse brain antigens for identi- fication. Antigens and antisera(prepared in mice given 6 weekly intraperitoneal inoculations of infec- tious mouse brain againstCSIRO 122 and MI 19334)were used in a range of tests againstthe Australian viruses or antiseraheld at the Queens- land Institute of Medical Research.The serolog- FIGURE1. Map of Macquarie Island, showing the ical techniquesused at Yale Arbovirus Research distribution of sites from which ticks were collected. Unit (Y ARU) were as described by Casals.4 The shadedarea shows the part of the island 200 meters or more in height. RESULTS A total of 123 adult ticks, 1,450 nymphs and Island were Royal Penguins (Eudyptes chryso- approximately 500 larvae were received for pro- lophus schlegeh), King Penguins (Aptenodytes cessing for virus isolation. Delays in transship- patagonica), and Rockhopper Penguins (Eu- ment resulted in the death of many ticks en route dyptes chrysocome). However, the ticks were or their overgrowth with fungus. Thirty-three found almost entirely in the areas associated with strains of virus, as shown in Table 1, were iso- Royal Penguins. The ticks collected, engorged or lated from 201 pools. The location of the 8 sites unengorged, were held for periods of I to 6 from which the infected ticks originated is illus- months at ambient temperature before being sent trated in Figure 1. alive or in liquid nitrogen for storage in vaporous Where ticks were inoculated into mice and cell liquid nitrogen until processing for virus isola- cultures in parallel, strains were isolated in both tion. systems in all 19 instances. Of the remaining 14 The unengorged ticks were individually iden- strains, 4 were isolated in mice and lOin cell tified and separated into pools of 5 to 10 adults cultures but parallel isolation was not attempted. or up to 50 nymphs or larvae hatched from eggs. Complement fixation tests segregated the 33 The preparation of the ticks for virus isolation, strains into 6 groups, 2 of which were typified the intracerebral inoculation of mice, prepara- by the viruses Nugget and Taggert.' Two groups tion of complement fixation (CF) antigen from were antigenically unrelated types from which positive mice, and serological techniques were CSIRO 122 and MI 19334 were selected as pro- as described by Doherty et al.! In addition, ap- totype strains. Three isolates were mixtures of proximately 75% of the prepared pools were in- strains of MI 19334 and Nugget, and 1 strain Paper 1.8

408 ST. GEORGE ET AL.

TABLE 1 Origin and identity ofstrains ofviruses isolated from Ixodes (C.) uriae collectedfrom Royal Penguin rookeries on Macquarie Island

90 Hurd Pt 1975

122 Nymphal PrecariousPt 1976 + + 127, 128,135 Larval PrecariousPt 1976 + + 129 Adult PrecariousPt 1976 + + 146 Nymphal NuggetsBeach 1977 +. + 273,274 Nymphal PrecariousPt 1979 + MIl 9334 Adult NuggetsBeach 1975 + * "Precarious Pt" 120, 145 Adult Lusitania Bay 1975 + + (MI 19334) 123 Nymphal Lusitania Bay 1975 + + 124,125 Nymphal PrecariousPt 1976 + + 126,142 Nymphal PrecariousPt 1976 + + 136,144 Adult Hurd Pt 1977 + + * 89 Adult Red River 1976 + Taggert 91 Adult Sellick Bay 1976 * + (MI 14850) 121,130 Adult Hurd Pt 1977 + + 137,138 Adult Hurd Pt 1977 + + 267 Adult Waterfall Bay 1979 * + 141,143 Adult Waterfall Bay 1979 * + 140 Adult Nuggets Beach 1977 + * Nugget (MI 14847) 268, 269, 270 Adult SoutheastBay 1979 + Mixture Nugget and "Precarious Pt" 92 Adult Sellick Bay 1976 + Unidentified .Isolation in the system not attempted. remains unidentified. The following character- positive finding being a low titer reaction to West istics are reported for the 2 prototypes. Nile virus. A neutralization test in mice showed that CSIRO 122 was clearly distinct from Sua- GadgetsGully virus (CSIRO 122) marez Reef virus, the only tick-borne flavivirus previously isolated in Australia (Table 2). Initially, tests were carried out with CSIRO CSIRO 122 was confirmed as a flavivirus at 122 and 127 in parallel. However, once the 2 Y ARU by CF and indirect immunofluorescence proved to be identical then only CSIRO 122 was tests, being more closely related antigenically to used in further tests and was sent to Y ARU. the tick-borne than to other flaviviruses. It was, Antigens from both CSIRO 122 and 127 vi- however, easily distinguishable from the avail- ruses had a haemagglutinin at a pH above 6.4 able strains. The results of some of the CF tests and which was optimal at 6.8. In each case the are shown in Table 3, and the neutralization tests hemagglutinin was inhibited by antisera to the comparing CSIRO 122 with Central European Australian tlaviviruses Saumarez Reef (CSIRO tick-borne encephalitis virus (CETBE) are shown 4) and Murray Valley encephalitis (MF.M 66) in Table 4. In addition, a hyperimmune mouse but not to the alphaviruses Sindbis (MRM 39) serum produced against CSIRO 122 gave nega- and Ross River (T48) viruses used in the initial tive reactions at a dilution of 1:8 with the fol- screen. Both antigens were negative in a com- lowing antigens: Banzi, dengue 2, dengue 4, Edge plement fixation test with antisera prepared Hill, Kadam, Kyasanur Forest Disease, Koko- against 2 strains of Saumarez Reef, Murray bera, Kunjin, Murray Valley encephalitis, Sa- Valley encephalitis, West Nile, Kokobera, Kun- boya, Sepik, St. Louis encephalitis, Usutu and jin, Edge Hill, Alfuy, Sepik, St. Louis encephalitis yellow fever viruses. and dengue (types I to 4) viruses with the only It is tentatively concluded that CSIRO 122 is F)aper 1.8

NEW ARBOVIRUSES AT MACQUARIE ISLAND 409

TABLE 2 TABLE 4 Antigen comparisonby HI, CF and neutralizationtests Identification a/strain CS-122-neutralization test by ofCSIRO 122strain with SaumarezReefvirus (SRE) IC route in mice using antisera against CSIRO 122 and Central European tick-borne encephalitis (CETBE) viruses Mouse ascitic fluid Virus SRE CSIRO 122 Virus CSIRO 122 CETBE Haemaggiutinationinhibiton -- Serum ICLD.. Log NI* Log NI* SRE 2:5,120 640 ICLDso CSIRO 122 160 320 Complementfixation* SRE 64/16 <8/<4 CSIRO 122 <8/<4 32/32 Neutralizationt SRE > 3.6 1.8 An homologous CF test was easily obtained CSIRO 122 0 1.6 with MI 19334 and after failing to find a reaction .Reciprocal serum titer/reciprocal antigen titer. with any antisera to Australian arbovirus anti- t Neutralization index in infant mice inoculated intracerebrally. gens and antisera the strain was sent to Y ARU. At YARU, mouse immune serum or ascitic a new tlavivirus more closely related to some of fluid for MI 19334 with homologous titers, the tick-borne complex than other tlaviviruses. 1:128-1 :256, were tested in dilutions beginning The small amount of serology carried out on at 1:4 or 1:8 against a large number of arbovirus Macquarie Island birds showed that 2 of30 Roy- antigens. Tests with the antigen prepared from al Penguin seracollected near Nuggets Beach (Fig. MI 19334 were carried out against a wide range 1) and 1 of 6 skua sera (Catharacta skua lonn- of grouping and specific sera, including mouse berg!) were reactive in a HI test with CSIRO 122 ascitic fluids prepared against 87 Bunyaviridae hemagglutinin. However, these same sera were (Table 6). As the results of both these sets of tests also reactive against a haemagglutinin prepared were consistently negative it is concluded that from Murray Valley encephalitis virus.. No neu- MI-19334 is an agent distinct from and unrelated tralization tests were done. to any of the recognized arboviruses, and is pre- sumably a new agent. MI 19334 was characterized as a bunyavirus- PrecariousPoint virus (MI 19334) like virus by electron microscopy. Thus it is pos- The physicalproperties of strain MI 19334are siblya new bunyavirus. However, a serological shownin Table 5. The haemagglutininwas active relationship has not been demonstrated with the at a wide pH range but at a low titer and inhi- known bunyaviruses. The identifying numbers bition tests were not possible. and origin of the other 9 strains which were found

TABLE 3 A comparison of CSIRO 122 virus with a range of tick-borne ./laviviruses by complement fixation

Serum

32+/32+ 16/32+ 32+/32+ 8/16 8/4 32+/32+ 32+/32+ 8/4 .Reciprocal of serum titer/reciprocal of antigen titer ..Central European tick-borne encephalitis. F)aper1.8

ST. GEORGE ET AL.

TABLE 5 Characteristicsof M/19334 isolatedfrom Ixodes (C.) uriae

Titers of virus pool (suspensionof third passage infected mouse brain) by: Intracerebral inoculation of infant mice* 8.3 Intraperitoneal inoculation of infant mice* 8.1 Intracerebralinoculation of weanedmicet negative Plaque assayon PS-EK cells:!: 8.4 Haemagglutinationof gooseerythrocytes at pH§11 6.0-6.6 Complement-fixation§~ 128/64 Sensitivity to sodium deoxycholate(1/1,000)** 6.1/2.0 Sensitivity to 50% diethyl ether** 2:7.5, <2.0 Titer (log,oLD,oIO.O15 ml) after filtration through membrane filter of averagepore diameter: 450 nm 6.7 300 nm 6.7 220 nm 6.7 100 nm 3.0 No reduction of titer with IUDR. Persistencebut not multiplication in inoculated Aedesaegypti. .Log,oLD,. per ml. t No evidence of paralysis at 10" infant mouse LD,o. * Loglo plaque-fonning units per ml. § Sucrose-acetone extracts of infected mouse brain as antigen. II Antigen diluted 1/10 and tested over pH range 6.0 to 7.6 at 37"C. 11Reciprocal serum titer/reciprocal antigen titer for homologous system. ..Log,oLD,. per 0.015 ml of virus suspension incubated without (as control) or with sodium deoxycholate or ether (4"C for 18 hr). to be identical to MI 19334 are listed in Table and Taggert virus at 5 sites additional to the 1. No serologicalsurvey was carried out with this single site which produced the original strains of virus. Nugget and Taggert demonstrates that these vi- ruses were not present only at the original site sampled on Macquarie Island. viruses The range and yield of viruses from ticks pro- Nugget virus was represented by 1 strain cessed in this series is substantial but still does (CSIRO 140)and 3 other strains of Nuggetvirus not reflect the full potential of the island. When were mixed with MI 19334 in tick collections the method of collection is considered over the which originated from a split single collection of time span covered, the number of ticks available ticks from SoutheastBay. Taggertvirus wasiso- for virus isolation was small. (Indirect methods lated from 9 pools of ticks collected at 4 sites of transport also were responsible for some loss- (Table 1). es.) However, tissue cultures were successfully The remaining strain CSIRO 92 was not iden- used to isolate strains of Gadgets Gully, Precar- tified, as an adequate homologous test system ious Point, Nugget and Taggert viruses. had not beendeveloped at the time the work on The strains of virus were isolated from 10(Co) this project was terminated. uriae collected on beaches on the eastern, western and southern coasts of the island. The other species of tick on Macquarie Island is Ixodes DISCUSSION kerguelenensis (synonym Ixodes pterodromae).5 The viruses isolated in this study originated It has been found only in the burrows of Dove from I. (C.) uriae ticks collected from sites oc- Prions (Pachyptila desolata) in upland areas6and cupied by colonies of Royal Penguins.The sites was not collected. were well spacedaround the shoresof the island One of the new viruses, CSIRO 122, is a fla- (Fig. 1). The isolation of Nugget virus at 1 site vivirus clearly distinct from the other flavivi- F)aper 1.8

NEW ARBOVIRUSES AT MACQUARIE ISLAND 41

TABLE 6 tributed in the subarctic (a.k.a. Ixodes (Ceratix- Virusesin the family Bunyaviridae testedby comple- odes)putus). CETBE virus has been associated mentfixation testswith Ml19334 mouseasciticfluid with Ixodes ricinus and Ixodes persulcatusand not with I. (C.) putus. I. (C.) putuswas associated with the type viruses of the Kemerovo and Anopheles A, Lukuni, Tacaiu- Sakhalin groupS.7-9Nugget and Taggert viruses ma are members of these antigenic groups Anopheles B AnophelesB respectively!so that a common invertebrate host Germiston, Guaroa, Ilesha, t Bunyamwera can be suggestedas a possible link betweenthe Kairi, Sororoca,Tensaw, subarcticand subantarcticin the biology of these Wyeomyia Bwamba Bwamba viruses, but so far no similar close link between Group C Apeu, Itaqui, Caraparu CETBE and CSIRO 122 has beendescribed. California California, Trivittatus, Melao The relationships found between I. (C.) uriae Capiro Capim, Guajara, Acara, Bush- of the far north and south of the world, and the bush Gamboa Gamboa viruses they carry are intriguing. Doherty et al.1 Guama Guama, Bertioga,Mirim, consideredthat the method of exchangeof ge- BeAn8438 I netic material of viruses and ticks betweenthe Koongol Koongol, Wongal subarctic and subantarctic regions could not be Nubututkab Nubututkab Oliflantsvlei Oliflantsvlei, Bobia explained by simple direct migration of partic- Patois Patois ular speciesof birds betweenthe land massesof Simbu Simbu, Akabane, Manzanilla, theseregions. However, in a review of tick-borne Nola, Utinga, Thimiri, Oro- viruses of seabirds,Clifford 10cites 2 speciesof pouche,Sathuperi, Buttonwil. seabirds which could fill such a role. Wilson's low Tete Tete, Bahig,Tsuruse Storm Petrel (Oceanitesoceanicus) breeds in the Turlock Turlock, Umbre, Yaba-l Antarctic and migrates to the Arctic. The recip- Sand fly fever Anhanga, Arumowot, Be- rocal breedingand migration pattern is followed AnlOOO49,Bujaru, Chagres, by the Arctic Tern (Sternaparadisaea). Itaporanga,Karimabad, Pa- cui, Salehabad,Naples, Sicil- Thus there may be a pathway whereby tick, ian, Punta Toro, Candiru, and perhaps viral genetic material may be ex- SudAn 754-61 changedbetween the subarctic and subantarctic Crimean-Congo CCHF, Hazara without it being establishedin the tropical zone hemorrhagic in an unmodified form. The generaldistribution fever Dera Ghazi Dera Ghazi Khan, Abu Mina of I. (C.) uriae is illustrated by Clifford.lo I. (C.) Khan uriaehas not beenrecorded betweenthe tropics. Hughes Hughes The closestthat I. (C.) uriae has been found to Nairobi sheep Dugbe the tropics wasa live female on a drowned Grey- disease headed Albatross (Diomedea chrysostoma), Qalyub Qalyub, Bandia Sakhalin CloMor, Taggert washedashore on Fraser Island, Australia (25°S Uukuniemi Uukuniemi, Grand Arbaud, latitude).!l EgAn 1825-61 Until the ticks of the genus Ixodes that occur Bakau Bakau in the temperate and tropical regions are more Kaisodi Silverwater, Lanjan thoroughly examined for Kemerovo, Sakhalin, Maputta Maputta Yogue Vogue CETBE virusesof the subarcticand Nugget,Tag- Unassigned Belmont, Bhanja, Kowanyama, gert and CSIRO 122viruses of the subantarctic, Lone Star, Tataguine,Witwa- a discontinuity of distribution of such closely tersrand,Keterah relatedviruses cannot be proven. There is scope for larger serologicalsurveys to determine what speciesof seabirdsare infected with theseviruses ruses known to exist in the Australian, or sub- to provide information on possibletransfer path- antarctic islands. However, there is a two-way ways. relationship between CSIRO 122 and CETBE of The bunyavirus MI 19334is more difficult to the northern hemisphere. The tick, 10 (Co) uriae categorizeas no serological relationships with of the southern hemisphere is also widely dis- known viruses were demonstrated,and no se- f)aper 1.8

412 ST. GEORGE ET AL. rological survey was carried out. Viruses of a lin group) from Ixodes uriae collected at Mac- number ofbunyavirus serogroupshave beeniso- quarie Island, Southern Ocean. Am. J. Trop. lated from ticks of the genus Ixodes,for instance Med. Hyg., 24:'521-526. 2. Serventy, D. L., Serventy, V., and Warham, J., of the Tete, CHF-Congo,Dera Ghazi Khan, Nai- 1971. The Handbook ofAustralian Sea-birds. robi SheepDisease, Sakhalin, Uukuniemi, Bhan- A. H. andA. W. Reed,Sydney. ja, Kaisodi and Thogoto serogroups.12The sig- 3. Macpherson,I., and Stoker, M., 1962. Polyoma nificance of MI 19334 as an infection or cause transformation of hamster cell clones-an in- vestigation of genetic factorsaffecting cell com- of diseaseof penguinsor other subantarcticver- petencte. Virology, 16: 147-151. tebrates must await future investigation. 4. Casals,J., 1979. Arenaviruses.Pages 815-841 in .\ This study has demonstrated that 2 viruses ,E. H. Lennette and N. J. Schmidt, eds., Diag- apparently new to science exist on Macquarie nostic Proceduresfor Viral. Rickettsial and Island. The names which are proposed are as- Chlamydial Infections. 5th edition. American Public Health Assn., Washington,DC. sociated with Macquarie Island history and are 5. Wilson, N. 1970. Acarina: Metastigmata: Ixo- attachedto geographicalfeatures near the points didae of South Georgia, Heard and Kerguelen. of collection. For the tick-borne flavivirus, Pac. Insects Monogr., 23: 78-88. CSIRO 122,we proposethe nameGadgets Gully 6. Murray, M. D., and Vestjens,W. J. M., 1967. Studies on the ectoparasitesof sealsand pen- and for the bunyavirus, MI 19334, Precarious guins. Ill. The distribution of the tick Ixodes Point. uriae White and the fiea Parapsyllusmagellan- A total of 4 tick-borne arboviruseshave now icus heardi de Meillon on Macquarie Island. beenfound on Macquarie Island. In particular, Aust. J. Zool., 15: 715-725. the new tick-borne flavivirus related to CETBE 7. Lvov, D. K., Timofeeva,A. A., Gromashevski,V. L., Chervonsky,V. I., Gromov, A. I., Tsyrkin, warrants evaluation as a possible human as well Y. M., Pogrebenko,A. G., and Kostyrko, I. N., as a bird pathogen. The total amount of work 1972. "Sakhalin" viru$-a new arbovirus iso- done on ticks and viruses from Macquarie Island lated from Ixodes (Ceratixodes)putus Pick.- is very small and clearly much more would be Camb. 1878collected on Tuleniy Island, Seaof Okhotsk. Arch. GesamteVirusforsch., 38: 133- justified. Macquarie Island is small in physical 138. size and isolated from large land massesso it 8. Lvov, D. K., Timopheeva,A. A., Gromashevski, may offer a most interesting ecosystemin which V. L., Tsyrkin, Y. M., Veselovskaya,O. V., Gos- to study virus ecologyin ticks and birds. tinshchikova, G. V., Khutoretskaya,N. V., Po- grebenko,A. G., Aristova, V. A., Sazonov,A. A., Chervonski,V. I., Sidorova,G. A., Fomina, ACKNOWLEDGMENTS K. B., andZhezmer,V. Y., 1973. "Okhotskiy" virus, a new arbovirus of the Kemerovo group We thank the National Parksand Wildlife Ser- isolated from Ixodes (Ceratixodes)putus Pick.- vice, Tasmania,and the Department of Health, Camb. 1878 in the Far East. Arch. Gesamte Virusforsch.,41: 160-164. Canberra,for permissionto collectticks on Mac- 9. Main, A. J., Downs, W. G., Shope, R. E., and quarie Island, and the Director, Antarctic Di- Wallis, R. C., 1973. Great Island and Bauline: vision, Department of Science,for assistancein Two new Kemerovo group from various ways; Dr. D. J. Griffiths, Dr. M. M. Bry- Ixodes uriae in easternCanada. J. Med. Ento- den, Dr. G. W. Johnstoneand Dr. I. R. Morgan mol., 10: 229-235. 10. Clifford, C. M., 1979. Tickborne viruses of sea- for collecting and transporting ticks; Mr. N. T. birds. Pages83-100 in E. Kurstak, ed., Arctic Hunt, Mr. S. S. Davis and Mr. G. J. Barrow for and Tropical Arboviruses.Academic Press,New technical expertise. York. II. Marks, E. N., 1968. Parasitesof a grey-headed albatrossfrom Ft'aserIsland. Queensl.Nat., 19: REFERENCES 30. 12. Chappell,W.. A., ed., 1984. -borne Vi- Doherty,R. L., Carley,J. G., Murray, M. D., Main, rus Information Exchange.Centers for Disease A. J., Kay, B. H., and Domrow, R., 1975. Iso- Control, Atlanta, Georgia. , lation of arboviruses(Kemerovo group, Sakha-