. . . . . . . . . . . . . . . . . . . . . . . . . . . Reports. . . . , . . . . . . . . . . . . . . . . . . . . . . .

Occurrence of Pacora virus (PAC: Bunyaviridae: Buunyavirus - like) in Brazilian Amazonia:

New .fmdings

~ I M L J N D O N.P. SOUTO', NICOLAS DEGALLIER~, AMÉLIA P.A. TRAVASSOS DA ROSA3, JORGE F.C. TRAVASSOS DA ROSA3

'Instituto de Pesquisas do Aiiiapú (IEPA), Macapú, AP 68900-310, 20RSTOM Belém, PA 6601 7-970, 'Iizstituto Evaridro Chagas (IEC), Fundação Nacional de Saúde, Ministério da Saúde, Serviço de Arbovírzis, Belém,

PA 66090-000, Brasil

The Pacora virus (PAC: Bunyaviridae: Bunyavirus - like), was known only through some iso- lates obtained from mosquitoes (Culex dunnì) in Panama, and two isolates obtained in the Bra- zilian Amazon region from the sylvatic birds Plzlegopsis nigronzaculata and Automolus oclzrolaemm. In order to add to the knowledge of the arboviruses' cycles, mosquitoes were col- lected in the Zoobotanical Park of Macapá (Amapá State), from August, 18th to September, l l th, in areas of rain forest and savannah. The collections were done at both ground and canopy levels in the forest and at ground level in the savannah, with human bait and light traps. 5642 mosquitoes have been collected, of which 5580 (or 184 pools) have been inoculated intracerebrally in newborn mice. Two strains of PAC virus have been isolated from unidentified Culex mosqui- toes. This result is the first report of PAC virus in Amapá State. The virus has also been isolated for the first time from mosquitoes in Brazil. Thus, the presence of the complete cycle of sylvatic transmission of this agent is confirmed in Brazilian Amazonia. The ecological data available for the hosts show that the virus circulates at the lower level in the rain forest (0-5 m) with mostly nocturnal mosquitoes as vectors.

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O virus PAC era conhecido apenas por al- g i n s isolainelitos obtidos no Patianid, a par- tir de niosquitos (Culex duniti) e de dois iso- lawientos obtidos iza Ainazônia brasileira-a partir de aves (PIdegogsis nigvoittnculntn e Autoniolus ochrolnemus). Coni objetivo de aprimorar os coiihecirneiitos sobre o ciclo silvestre de transmissão dos arbovirus foravi realizadas coletas de niosquitos no Parque Zoobotânico de Macapú (AP), no periodo de

18/08 cÌ 11/09/92, ein uma úrea de floresta de ferra firme e ein uma área de savaiia. As coletas forain realizadas a nivel do solo e da copa da floresta, e somente a nivel do solo na savana, utilizando-se simultaneainente iscas humana e luminosa. De um total de 5640 inosqiiitos coletados, 5580 (184 lotes) foram inoculados intracerebralinente en2 ca- mundongos recém-nascidos. Dois isolamen- tos do vírus PACforam obtidos de dois lotes

coiistituídos de 50 inosquitos indeterminados do gênero Culex. Além do primeiro registro da presença do virus PAC no Estado do Amapá, os presentes achados confirniani a existêiicia do ciclo cornpleto de transmissão do virus na Ainazôriia brasileira. Os dados ecológicos disponíveis iiidicain que este ví- rus circula deritro dos estratos baixos (0-5 m) na floresta de terra firme, transniitido por niosquitos notiirnos.

he Pacora virus (PAC: isolations made from wild birds Bunyaviridae: Bunyavirus - (Plilegopsu nigromaculata, Fosmica- like), distantly related to the riidae and Autoniolus ochrolaemus, antigenic group Simbu (I), is Furnariidae) collected in Brazil (3). Correspondence to. Ralmundo N.P Souto, Ins-

na1 de Saúde, Ministério,da Saúde, Serviço de known by some strains isolated from These species of birds are dwelling pref- mosquitoes of the species CuZex durzryi erentially in terrafirlriie forests, at low Arbovirus, postal Ilz8, Belém, 66090-

collected in Panaina (2), and by two level (0-5 in) (2) and CuZex mosquitoes 000, Brasil.

T Chagas (IEC),

Volume 48(4) * July/August 1996 Ciência e Cultura Journal of the Brazilian Association for the Advancement of Science 261 _ - -~

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are generally nocturnal and omithophilous in habits (4). However, the existence of the transmission cycle of PAC virus in Brazil had not been defi- nitely established. ,k ~ &,

As a part of a wider study entitled “A comparative study of the seasonality of mosquitoes (Diptera, Culicidae) and isolation attempts of arboviruses in the rain forest and savannah of Amapá State, Brazil”, this paper will present new data about the cycle of this arbovirus.

The study has been conducted in the Zoobotanical Garden, located in the district of Fazendinha, and situated on the road Macapá - Santana, 1 O km fi-om Macapá (5). The climate is classified as tropical humid, with a mean annual pre- cipitation of 2500 mm and a mean an- nual temperature of about 27°C (6,7). The area consists of a patch of old sec- ondary rain forest bordered by an open area of savannah (8).

Adult mosquitoes have been col- lected simultaneously in the savannah and the forest, and in the latter at ground level and in the canopy. Anthropo- philous species have been collected as they were landing on human bait and others with light traps (9,lO). The sam- ples have been conserved and processed following standard entomological (1 1) and virological methods (1 2).

From August, 18 to September, 11, 1992 a total of 5642 mosquitoes, ac- counting for at least 36 species and 13 genera, have been collected. Of these, 5580 (1 84 pools) have been inoculated intracerebrally in newborn mice (Table 1). Two strains of PAC virus (13) have been isolated from the 36 pools (1722 species) made of unidentified Culex mosquitoes. Despite the fact that the latter represented 30.8% ofthe total, the minimum infection rate (MIR) for PAC virus was fairly low (0.1 1%; 2/1722), suggesting a low level of transmission.

Since PAC virus is recorded for the first time’ in Amapá State and fi-om mos- quitoes in Brazil, its complete sylvatic cycle is confirmed to occur in Brazilian Amazonia. As suggested by the ecologi- cal preferences of the known hosts, this virus has probably a bird-Culex cycle, mainly at ground level in the forest and

Table 1 - Mosquitoes collected in the Zoobotanical Park, Macapá, Amapá State, Brazil, Au- gust, 18 - September, 11, 1992, and inoculated for arbovirus isolation; the positive species are in boldface.

Species Collected Inoculated Pools

Culicidae Anophelinne An. (Anopheles) sp. An. (Nj~ssorliynch~rs) sp. An. (Nss.) hi-aziliensis An. (í‘$u.) nimeztovari An. (Nia.) oswaldoi An. (Nys.) triannidatirs

Culicinae A edeoniyiini A edeornyin (iiedeoniyia) squaniipennis Aedini Ae. (Ochlerotatirs) scapirlaris Ae. (Och.) serratus Psorophora (Grabhanzia) cingidata Ps. (Janthinosoiua) albipes Ps. (Jan.) .firox Crrlicini

Culer sp. Cx. (Czrlex) sp.

Cx. (C1N.J coronator G. (Cia.) corniger

Cx. (Crrx.) declarator Cx. (Melanoconion) sp. Cx. (Mel.) adaniesi Cx. (filel.) pedroi Cx. (Mel.) spissipes Cx, (Mel.) ocellatirs Cx. (filel.) portesi C.X. (filel.) vomerifer fiíansoniini Cq, (RhjaiChotaeilia) nigricans Cq. (Rhy.) venezuelensis Mansonici (Mansonia) sp. Ma. (Man.) hicnieralis &ïu. (ManJ amazonensis Ma. (Man.) pseirdotitillans Ma. (Man.) titillans Sabethini Linzafiis sp. Li. dzrrhaniii Phoniornyia sp. Sabethes (Sabethes) belisurioi Sa. (Sub.) cyaneirs Sa. TSab.) qirasicyaneirs Sa. (Sob.) tarsopiis Sa. (Sabethoides) glaucodaenion Pichoprosopoli digitatinn @eoniyia sp. Urariotaeniini Uranotaenia sp. UK hystera .

10 210 830 12 3

670

515

26 69 2

108 400

1722 13 1

33 17 22 1 6

18 1 1 1

1 310 95 6 2

20 313

1 4 9 5 1 9 1

12 17 57

76 12

5 208 829

5

670

514

24 67

104 398

1122 13

28 17 17

18

310 95 6

20 313

1 I 6 5 1 9 1

12 17 57

76 11

1 6

21 1

17

19

2 4

4 17

36 1

2 2 1

2

10 4 1

1 14

1 1 1

3 1

Total 5642 5580 184

nocturnal, following the habits of its mosquito vectors which are nocturnal and ornithophilous in habits (4).

At the present time, it is difficult to speculate about the causes of the dis- junct geographical distribution of this virus, which has been found only in

Central America and Amazonia. How- ever, as some authors have already noted, arboviruses with mosquito-bird cycles are in general widely distributed (2,14,15), probably due to the migrat- ing habits of some of their hosts (1 6,17,1 8). Among others, Saint Louis

262 Ciência e Cultura Journal of the Brazilian Association for the Advancement of Science Volume 48(4) July/August 1996

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Encephalitis (SLE), Eastern and West- ern Equine Encephalitis (EEE and WEE, respectively), Ilheus (ILH) and Gainboa (GAM), and Una (UNA) vi- ruses have been found from North America (or at least Central America) to Argentina (19,20) and may also be found in the Guianas and Brazil. Turlock virus (TUR) is found from North America to Brazil (21). Thus it inay well be possible that, although not being so extensively distributed, the PAC virus has been dispersed by its avian hosts.

References and notes

1. Zeller HG, N Karabatsos, CH Calisher, JP Digoutte, CB Cropp, FA Murphy, RE Shope 1989 Electron microscopic and antigenic studies of uncharacterized viruses. II. Evi- dence suggesting the placement of viruses in the family Bunyaviridae. Aidi Vilal 108: 21 1- 227

2.Galindo APMC 1978 Los arbovirus de Panamá. Rev MEd Pai7amá 3: 1-41

3. Degallier N, APA Travassos da Rosa, JMC Silva, SG Rodrigues, PFC Vasconcelos, JFS Travassos da Rosa, GP Silva, RP Silva 1992 As aves como hospedeiros de arbovirus na Amazônia brasileira. Bol Mus Para Eiizilio Goeldi sé/* Zoo1 8: 69- 11 1

4. Mattingly PF 1960 Ecological aspects of the evolution of mosquito-borne virus diseases. Trans R Soc Pop Med Hyg 54: 97-1 12

5. Altitude: 14 m above see level and geographic coordinates: 00”02’S, 51’03’W

6. According to the Koppen’s classification, the Af climate is characterized by (i) the drier month with 60 mm or more rain, and (ii) the annual average of the maximum and mini- mum temperatures being 3 1°C and 23”C, re- spectively; the drier and wetter quarters are September-November and March-May, re-

spectively; annual mean air relative humidity is 85% and the mean annual sun time is 2200 hours

7. Sudam 1984 Atlas cliii7atológico da Ama- z6i7ia brasileira. Projeto de Hidrologia e Climatologia da Amazônia. Publ. 39, Belém

8. Its extension is about one kin wide by two kin depth between the road and a lake in the sa- vannah

9. Service MW 1976 Mosqzrito ecology. Ap- plied Science Publishers Ltd, London

1 0 . h the forest, at ground level and on a plat- form at 15 in, and in the savannah at ground level, the human baited collecting sessions were conducted from 1O:OO li to 13:OO 11 and from 16:OO 11 to 19:OO li; at each location, a “CDC miniature light-trap” was operating from 18:OO 11 to 6:OO h

11. In the field, the samples were conditioned in glass tubes for storage in liquid nitrogen (-196°C); in the laboratory, the sorting of the mosquitoes was done above a chilling table (-20°C) and monospecific pools of 50 specimens each were conserved in a freezer (-70°C) until further processing

12.After being ground, each pool was diluted in bovine albumin (0.75%) with antibiotics, centrifuged at 2000 rpm for 10 min; the supernatant was then inoculated intracere- brally into 6 newborn mice (0.02 ml/animal), which were observed daily during three weeks for any sign of disease; if this was the case, further passages andlor serological tests were done with the brain of the sick animals. The identification ofthe strains was done by the complement fixation test, accord- ing to Fulton and Dumbell (1946 J Gen Microbrol3: 97-l l l ) , and using ascitic flu- ids hyperimune against known Amazonian arboviruses

13. Strains AR 517443 and AR 5 17452 14. Calisher CH, N Karabatsos, JS Lazuick, TP

Monath, KL Wolff 1988 Reevaluation of the Western Equine Encephalitis antigenic complex of alphaviruses (family Toga- viridae) as determined by neutralization tests. A m J Trop Med Hl~g 38: 447-452

15. Calisher CH 1988 Evolutionary signifi- cance of the taxonomic data regarding

. . . . . . . . . . . . . . .

2

~-

. . . . . .

Bunyaviruses of the family Bunyaviridae. Iizìervirology 29: 268-276

16.Calisher CH, ICSC Maness, RD Lord, PH Coleman 1971 Identification of two South American strains of eastern equine encephaloinyelitis virus from migrant birds captured on the Mississippi delta. Ain J Epideriiiol 94: 172- 178

17.Calisher CH, VE Gutiérrez, RD Lord, KSC Maness 1975 Aislamento de virus Mayaro de un ave migratoria capturada en Luisiana en 1967. Bol Of Sail Pniiaiizer 78: 148-154

18.Dickerinan RB, MS Martin, EA Dipaola 1980 Studies of Venezuelan Encephalitis in migrating birds in relation to possible transport of virus from South to Central America. Ain J Pop Med Hyg 29: 269-276

19. Karabatsos N 1985 Znterizatioiial catalogue of arboviruses iiiclztding certain other, vi- ruses of verfehates. Third ed, American Society of Tropical Medicine and Hygiene, San Antonio, USA

20.Calisher CH, JS Lazuik, G Justines, DB Francy, TP Monath, VE Gutierrez, MS Sabattini, GS Bowen, WL Jakob 1981 Vi- ruses isolated from Aedeoinyia sqzramipeiinis mosquitoes collected in Panama, Ecuador, and Argentina: establish- ment of the Gamboa serogroup. Ain J Trop MedHvg 30: 219-223 Degallier N, APA Travassos da Rosa, PFC Vasconcelos, JP Hervé, GC Sá Filho, JFS Travassos da Rosa, ES Travassos da Rosa, SG Rodrigues 1992 Modifications of arbovirus transmission in relation to con-

, struction of dams in Brazilian Amazonia. Ci Cult JBraz Assoc Adv Sci 44: 124-135

22.Acknoi~ledgen2ents: This work has been done with the financial and logistical sup- ports of the following institutions: IEPA (Macapá, Amapá), ORSTOM (Paris, França), IEC-FNS-MS (Belém, Pará), MPEG/UFPA (Belém, Pará) and CNPq (Brasília, DF). The technical staff of the en- tomological section in IEC has helped us during the identification of the mosquitoes.

Received 20 April 1994 Accepted O1 September 1994

Volume 48(4) July/August 1996 Ciência e Cultura Journal of the Brazilian Association for the Advancement of Science 263

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Seasonal shift in food resources affec

I

Setor de Ecologia, Instifirto de Biologia, Un idade do Estado do Ri .o, RJ ,70550-000, Brasil

The diet of the tropidur habitat of the Restinga Brazil. The diet comp season (winter) and r that in both seasons the diet w in terms of proportio dance of arthropods ably lower being on

cantly related to their availa 13; p < 0.001 for d Ipomoea litorallis fl flowers in the envi observed for arthropods, higher during the

that of the wet se that shifts season

selectiveness. T

iro State, southeastern the wet (summer) and dry

s but differed significantly 268; p < 0.001). The abun- ropods/m2) was consider- (x = 5.4 arthropods/m').

son). The proportion of ntly related to the availability of

ropods consumed were signifi- rank correlation, rs= 0.87; N =

emis liitzae has a to consume arthr

3.78) was 68% bro

A composição da dieta do lagarto trop Liolneiiiiis htzne foi estirdada em irni ca zonal na Restinga da Barra de Maricá, pio de Maricá, Rio de Janeiro, sirdeste sil. A dieta do lagarto foi comparada entre as estações de chuva (verão) e seca (inverno), sen- do relacionada coni a disponibilidade de recur- sos alimentares no ambiente. Os dados mostra- ram qire em ambas as estações a dieta estava coniposta por itens de origem animal e vegetal nias d$erindo signijicativamente em termos das proporções consiiniidas (Kolniogolav-Si~~ir~io~~ test, Dmny= 0.268: p < 0.001). A abundância de artrópodos no unibiente na estação seca (x = 2,l artrópodos/ni2) era apenas 39.704 daqirela me-

ra relativa do nicho alimen- ctznc rlurante a estação seca

maior do qire a estimadapara

este lagarto tende a consir-

plantas ingeridas provavelmente possireni um papel importante não apenas conio reciirso ali- nientai; mas tumbe" como fonte de ágm.

W Correspondence to: Carlos Frederico Duarte Rocha, Setor de Ecologia, Instituto de Biologia, UERJ, Rua São Francisco Xavier, 524, Maracanã, Rio de Janeiro, RJ 20550-000, Brasil.

k

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264 Ciência e Cultura Journal of the Brazilian Association for the Advancement of Science Volume 48(4) July/August 1996

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