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Rickettsia bellii in ticks Amblyomma varium Koch, 1844, from birds in Peru
Ticks and Tick-borne Diseases 3 (2012) 254–256
Contents lists available at SciVerse ScienceDirect
Ticks and Tick-borne Diseases journal homepage: www.elsevier.de/ttbdis
Short communication
Rickettsia bellii in ticks Amblyomma varium Koch, 1844, from birds in Peru Maria Ogrzewalska a, Ivan Literak b,c,∗, Jorge M. Cardenas-Callirgos d, Miroslav Capek e, Marcelo B. Labruna a a
Departamento de Medicina Veterinaria Preventiva e Saude Animal, Faculdade de Medicina Veterinaria e Zootecnia, Universidade de Sao Paulo, Sao Paulo, Brazil Department of Biology and Wildlife Diseases, Faculty of Veterinary Hygiene and Ecology, University of Veterinary and Pharmaceutical Sciences Brno, Brno, Czech Republic CEITEC VFU, University of Veterinary and Pharmaceutical Sciences Brno, Brno, Czech Republic d Laboratorio de Invertebrados, Museo de Historia Natural, Facultad de Ciencias Biologicas, Universidad Ricardo Palma, Lima, Peru e Institute of Vertebrate Biology, Academy of Sciences of the Czech Republic, v. v. i., Brno, Czech Republic b c
a r t i c l e
i n f o
Article history: Received 30 March 2012 Received in revised form 11 May 2012 Accepted 23 May 2012 Keywords: Rickettsia bellii Ticks Amblyomma calcaratum Birds Peru
a b s t r a c t Amazonian birds were caught and examined for the presence of ectoparasites in the Allpahuayo Mishana National Reserve near Iquitos, Peru, from 13 to 16 August 2011. A total of 40 birds representing 16 species were examined. Two birds (5%) were infested with 2 larvae of Amblyomma varium Koch, 1844, and one nymph of A. calcaratum Neumann, 1899. The 2 larvae of A. varium were infected with Rickettsia bellii. This is the first report of R. bellii in A. varium and also the first record of this rickettsia in Peru. In addition, an immature A. calcaratum is reported from Peru for the first time. © 2012 Elsevier GmbH. All rights reserved.
Introduction
Materials and methods
Information on the tick fauna and tick-borne pathogens in Peru is scarce. The presence of 50 tick species (35 of the family Ixodidae, 15 of Argasidae) has been confirmed in Peru (Need et al., 1991; Caceres et al., 2002; Mendoza-Uribe and Chaves-Chorocco, 2004; Blair et al., 2004; Guglielmone et al., 2005; Labruna et al., 2005, 2010; Nava et al., 2010). Wild birds are commonly parasitized by larvae and nymphs of ticks, and they are undeniably among tick hosts with the highest mobility, and therefore they may be regarded as hosts with the highest potential for dispersal of ticks and tick-borne diseases (Hoogstraal, 1961). Bacteria of the genus Rickettsia are obligate intracellular organisms that infect invertebrate hosts worldwide. Some of them cause disease (i.e., rickettsioses) in warm-blooded animals and humans, to whom they are transmitted by hematophagous vectors, mostly ticks (Raoult and Roux, 1997). The present study notes tick infestations on wild birds in the Amazonian forest of Peru and rickettsial infection proved in these ticks.
Mist nets were used to trap wild birds in Peru in summer 2011. Birds trapped were subjected to examination for the presence of ectoparasites including ticks. The ticks were collected after naked eye examination using a tweezer and preserved in 96% ethyl alcohol. After examination, all the birds were released back into the wild. Four study sites were as follows: (i) Refugio de Vida Silvestre Los Pantanos de Villa, Lima, 12◦ 13 S, 76◦ 59W, at sea level (a.s.l.), 50 birds of 10 species were examined from 19 to 20 July 2011. (ii) Centro URKU, Tarapoto, 06◦ 27 S, 76◦ 21 W, 410 m a.s.l., 39 birds of 21 species were examined from 8 to 10 August 2011. (iii) Reserva Nacional Allpahuayo Mishana, Iquitos, 03◦ 58 S, 73◦ 25 W, 134 m a.s.l., 40 birds of 16 species were examined from 13 to 16 August 2011. (iv) Cascay, Huanuco, 09◦ 50 S, 76◦ 08 W, 1845 m a.s.l., 104 birds of 18 species were examined from 20 to 23 August 2011. A total of 233 birds of 57 species were examined (Table 1). The birds were identified using the field guide by Schulenberg et al. (2010). Nymphal ticks were identified according to Martins et al. (2010). To identify larvae, we used molecular tools as described elsewhere (Ogrzewalska et al., 2011). Larvae collected were individually submitted to DNA extraction and then to PCR using primers targeting the tick mitochondrial 16S rDNA gene (Mangold et al., 1998) and the 12S rDNA gene (Beati and Keirans, 2001). Amplified products were purified and DNA-sequenced as previously described (Labruna et al., 2004a) and compared with NCBI Nucleotide BLAST searches (Altschul et al., 1990).
∗ Corresponding author at: Department of Biology and Wildlife Diseases, Faculty of Veterinary Hygiene and Ecology, University of Veterinary and Pharmaceutical Sciences Brno, Palackeho 1-3, 612 42 Brno, Czech Republic. E-mail address: [email protected] (I. Literak). 1877-959X/$ – see front matter © 2012 Elsevier GmbH. All rights reserved. http://dx.doi.org/10.1016/j.ttbdis.2012.05.003
M. Ogrzewalska et al. / Ticks and Tick-borne Diseases 3 (2012) 254–256 Table 1 A list of birds examined (PV, Refugio de Vida Silvestre Los Pantanos de Villa, Lima; TR, Centro URKU, Tarapoto; IQ, Reserva Nacional Allpahuayo Mishana, Iquitos; HU, Cascay, Huanuco). Family/species Columbidae Columbina cruziana Columbina minuta Leptotila rufaxilla Leptotila verreauxi Trochilidae Amazilia amazilia Amazilia fimbriata Amazilia chionogaster Amazilia lactea Doryfera johannae Glaucis hirsutus Phaethornis atrimentalis Phaethornis superciliosus Thalurania furcata Threnetes leucurus Cuculidae Crotophaga ani Ardeidae Butorides striata Ixobrychus exilis Laridae Larus belcheri Bucconidae Bucco macrodactylus Falconidae Falco sparverius Thamnophilidae Thamnophilus schistaceus Furnariidae Dendrocincla fuliginosa Glyphorhynchus spirurus Phleocryptes melanops Pipridae Lepidothrix coronata Machaeropterus pyrocephalus Manacus manacus Pipra fasciicauda Cotingidae Pachyramphus polychopterus Tyrannidae Elaenia spectabilis Leptopogon amaurocephalus Mionectes oleagineus Pitangus lictor Pitangus sulphuratus Sayornis nigricans Tachuris rubrigastra Todirostrum cinereum Tolmomyias flaviventris Tyrannus melancholicus Vireonidae Vireo olivaceus Hirundinidae Pygochelidon cyanoleuca Troglodytidae Campylorhynchus fasciatus Troglodytes aedon Polioptilidae Ramphocaenus melanurus Turdidae Turdus ignobilis Fringillidae Carduellis magellanica Emberizidae Zonotrichia capensis Cardinalidae Pheucticus chrysogaster Thraupidae Diglossa sittoides Oryzoborus angolensis Ramphocelus carbo Sicalis luteola
No. of birds examined 2 7 3 1 1 1 19 2 1 5 4 10 1 3
Locations HU HU IQ HU PV IQ HU TR TR TR (2), IQ (3) TR TR (5), IQ (5) IQ TR
4
HU
2 2
PV PV
1
PV
1
TR
2
PV (1), HU (1)
2
TR
3 5 9
IQ IQ PV
4 3 1 2
IQ TR (2), IQ (1) IQ TR
4
TR (2), IQ (2)
1 1 7 1 1 3 23 1 1 2
TR TR TR (1), IQ (6) IQ TR HU PV HU TR HU
2
TR
2
HU
6 7
HU PV
1
TR
2
TR (1), HU (1)
255
Table 1 (Continued) Family/species
No. of birds examined
Locations
Sporophila nigricollis Sporophila telasco Tachyphonus luctuosus Thraupis episcopus Tiaris obscurus Total
9 1 1 19 3 233
HU PV TR HU TR
Two larvae were individually tested for the presence of Rickettsia by PCR targeting a fragment of the gltA gene, which occurs in all Rickettsia species (Labruna et al., 2004a), and another PCR targeting a fragment of the ompA gene that occurs only in Rickettsia species of the spotted fever group (SFG). PCR products were DNA-sequenced and submitted to BLAST analysis to determine similarities to other Rickettsia species. Results The ticks were found only on Amazonian birds in the Reserva Nacional Allpahuayo Mishana where 40 birds belonging to 16 species were captured. Two birds (5%) were infested with ticks: one of 3 grey-fronted doves Leptotila rufaxilla (Columbiformes: Columbidae), and one of 2 bicolored antbirds Gymnopithys leucaspis (Passeriformes: Thamnophilidae). Two larvae of Amblyomma sp. were found on L. rufaxilla. Their 16S rDNA sequences were identical to each other, and 88.6% (364/411 bp) similar to A. brasiliense (FJ424399) and 87.6% (361/412 bp) to A. maculatum (L34318). Their 12S rDNA sequences were identical to each other and 97.6% (331/339 bp) similar to A. varium (AY974339) from Brazil and 99.4% (313/315 bp) to A. varium (AY342283) from Peru. As there is no specimen of A. varium available in GenBank, the found larvae were identified as A. varium based on the high similarity of the sequences of the 12S rDNA fragment to that species. Based on the gltA gene PCR, these 2 A. varium larvae were found infected with a Rickettsia 99.1% (333/336) similar to R. bellii strain Pontal, previously isolated from A. nodosum in Brazil (EU567181), and R. bellii strain HJ7 (DQ865204) isolated from Haemaphysalis juxtakochi Cooley, 1946, also in Brazil. This high similarity (>99%) of the gltA partial sequences is indicative of that these 2 Amblyomma larvae from Peru were infected by R. bellii. The samples were negative to PCR targeting the ompA gene, which was expected because R. bellii is not an SFG species. One nymph of A. calcaratum Neumann, 1899, was found on G. leucaspis. The nymph was not tested for the presence of rickettsia, and it was deposited at the tick collection “Colecao Nacional de Carrapatos” (CNC) of the University of Sao Paulo, Brazil (accession number 2155). GenBank nucleotide sequence accession numbers for the partial sequences generated in the present study are JQ771810 for the R. bellii partial gltA sequence, and JQ771811 and JQ771812 for the mitochondrial 12S rDNA and 16S rDNA gene of the A. varium larva, respectively. Discussion
14
HU
5
HU
5
HU
3 3 1 2
HU IQ IQ PV
A. calcaratum has been reported in various countries of Central and South America (Guglielmone et al., 2003). However, this is the second report of A. calcaratum in Peru and the first report of an immature stage of A. calcaratum on a bird in this country. The adult stage parasitizes chiefly anteaters, Tamandua spp. and Myrmecophaga tridactyla, whilst immatures feed mostly on wild passerine birds (Jones et al., 1972; Ogrzewalska et al., 2011). Until now, the known presence of A. calcaratum in Peru has been restricted to 12 adult specimens collected from the southern tamandua Tamandua tetradactyla in the Caserio Menor de Nunhan Jalca, Fundo Santa
256
M. Ogrzewalska et al. / Ticks and Tick-borne Diseases 3 (2012) 254–256
Rosa, Bagua Grande, Utcubamba Amazonas (Caceres et al., 2002). Here, we add one new host species for A. calcaratum, G. leucaspis, which has not previously been reported to be parasitized by this tick species. A. varium is a neotropical tick, known as the sloth’s giant tick, with records from southern Central America to Argentina, including Peru (Guglielmone et al., 2003). Adults are found almost exclusively on mammals of the families Bradypodidae and Magalonychidae (Xenarthra) (Guglielmone et al., 2003; Need et al., 1991), however, the hosts of the immature stages have been still unknown, so we speculate that their hosts could be birds. In this study, both A. varium larvae were infected with R. bellii. R. bellii has been notified as a common Rickettsia in various species of hard ticks from Brazil (Labruna et al., 2004a, b, 2007a; Pinter and Labruna, 2006; Ogrzewalska et al., 2009; Pacheco et al., 2008) and Argentina (Labruna et al., 2007b; Tomassone et al., 2010); however, there is no evidence that it is pathogenic for humans or animals (Labruna, 2009; Labruna et al., 2011). This is the first report of R. bellii in Peru and the first finding of this rickettsia infecting A. varium. Acknowledgments We thank Martina Literakova, Zuzana Literakova, and Blgo. Kember Mejia Carhuanca (Director Programa de Investigacion en Biodiversidad Amazonica, Instituto de Investigaciones de la Amazonia Peruana, Iquitos, Peru) for their cooperation in the field. The field work was covered by a permission from responsible Peruvian authorities licensed for Lic. Mercedes Gonzales de la Cruz (Museo de Historia Natural, Facultad de Ciencias Biologicas, Universidad Ricardo Palma, Lima, Peru). The study was supported in part by the Grant Agency of the Academy of Sciences of the Czech Republic (grant IAA601690901) and by the Ministry of Education, Youth and Sports of the Czech Republic (grant LC06073). Ivan Literak was supported by the project ‘CEITEC – Central European Institute of Technology’ (CZ.1.05/1.1.00/02.0068) of the European Regional Development Fund. References Altschul, S.F., Gish, W., Miller, W., Myers, E.W., Lipman, D.J., 1990. Basic local alignment search tool. J. Mol. Biol. 215, 403–410. Beati, L., Keirans, J.E., 2001. Analysis of the systematic relationships among ticks of the genera Rhipicephalus and Boophilus (Acari: Ixodidae) based on mitochondrial 12S ribosomal DNA gene sequences and morphological characters. J. Parasitol. 87, 32–48. Blair, P.J., Jiang, J., Schoeler, G.B., Moron, C., Anaya, E., Cespedes, M., Cruz, C., Felices, V., Guevara, C., Mendoza, L., Villaseca, P., Sumner, J.W., Richards, A.L., Olson, J.G., 2004. Characterization of spotted fever group rickettsiae in flea and tick specimens from northern Peru. J. Clin. Microbiol. 42, 4961–4967. Caceres, A.G., Beati, L., Keirans, J.E., 2002. First evidence of the occurrence of Amblyomma calcaratum Neumann, 1899 in Peru. Rev. Peru Biol. 9, 116–117. ˜ A., Keirans, J.E., Robbins, R.G., 2003. Ticks (Acari: Guglielmone, A.A., Estrada-Pena, Ixodida) of the Neotropical Zoogeographic Region. International Consortium on Ticks and Tick-borne Diseases, Atlanta, Houten, The Netherlands, p. 173. Guglielmone, A.A., Romero, J., Venzal, J.M., Nava, S., Mangold, A.J., Villavicencio, J., 2005. First record of Haemaphysalis juxtakochi Cooley, 1946 (Acari: Ixodidae) from Peru. Syst. Appl. Acarol. 10, 33–35.
Hoogstraal, H., 1961. Migrating birds and their ectoparasites in relation to disease. East Afr. Med. J. 38, 221–226. Jones, E.K., Clifford, C.M., Keirans, J.E., Kohls, G.M., 1972. The ticks of Venezuela (Acarina: Ixodoidea) with a key to the species of Amblyomma in the western hemisphere. Brigham Young Univ. Sci. Bull. Biol. Ser. 17, 1–40. Labruna, M.B., Whitworth, T., Horta, M.C., Bouyer, D.H., McBride, J.W., Pinter, A., Popov, V., Gennari, S.M., Walker, D.H., 2004a. Rickettsia species infecting Amblyomma cooperi ticks from an area in the state of Sao Paulo, Brazil, where Brazilian spotted fever is endemic. J. Clin. Microbiol. 42, 90–98. Labruna, M.B., Whitworth, T., Bouyer, D.H., McBride, J.W., Camargo, L.M.A., Camargo, E.P., Popov, V., Walker, D.H., 2004b. Rickettsia bellii and Rickettsia amblyommii in Amblyomma ticks from the state of Rondônia, Western Amazon, Brazil. J. Med. Entomol. 41, 1073–1081. Labruna, M.B., Keirans, J.E., Camargo, L.M.A., Ribeiro, A.F., Soares, R.M., Camargo, E.P., 2005. Amblyomma latepunctatum, a valid tick species (Acari: Ixodidae) long misidentified with both Amblyomma incisum and Amblyomma scalpturatum. J. Parasitol. 91, 527–541. Labruna, M.B., Pacheco, R.C., Richtzenhain, L.J., Szabo, M.P., 2007a. Isolation of Rickettsia rhipicephali and Rickettsia bellii from ticks Haemaphysalis juxtakochi in the state of Sao Paulo, Brazil. Appl. Environ. Microbiol. 73, 869–873. Labruna, M.B., Pacheco, R.C., Nava, S., Brandao, P.E., Richtzenhain, L.J., Guglielmone, A.A., 2007b. Infection by Rickettsia bellii and Candidatus “Rickettsia amblyommii” in Amblyomma neumanni ticks from Argentina. Microb. Ecol. 54, 126–133. Labruna, M.B., 2009. Ecology of Rickettsia in South America. Ann. N. Y. Acad. Sci. 1166, 156–166. Labruna, M.B., Romero, M., Martins, T.F., Tobler, M., Ferreira, F., 2010. Ticks of the genus Amblyomma (Acari: Ixodidae) infesting tapirs (Tapirus terrestris) and peccaries (Tayassu pecari) in Peru. Syst. Appl. Acarol. 15, 109–112. Labruna, M.B., Mattar, S., Nava, S., Bermudez, S., Venzal, J.M., Dolz, G., Abarca, K., Romero, L., de Sousa, R., Oteo, J., Zavala-Castro, J., 2011. Rickettsioses in Latin America, Caribbean, Spain and Portugal. Rev. MVZ Córdoba 16, 2435–2457. Mangold, A.J., Bargues, M.D., Mas-Coma, S., 1998. Mitochondrial 16S rDNA sequences and phylogenetic relationships of species of Rhipicephalus and other tick genera among Metastriata (Acari: Ixodidae). Parasitol. Res. 84, 478–484. Martins, T.F., Onofrio, V.C., Barros-Battesti, D.M., Labruna, M.B., 2010. Nymphs of the genus Amblyomma (Acari: Ixodidae) of Brazil: descriptions, redescriptions, and identification key. Ticks Tick-Borne Dis. 1, 75–99. Mendoza-Uribe, L., Chaves-Chorocco, J., 2004. Amplicacion geografica de siete especies de Amblyomma (Acari: Ixodidae) y primer reporte de A. oblongoguttatum Koch, 1844 para Peru. Rev. Per. Ent. 44, 69–72. Nava, S., Velazco, P.M., Guglielmone, A.A., 2010. The first record of Amblyomma longirostre (Koch, 1844) (Acari: Ixodidae) from Peru, with a revision of its host range. Syst. Appl. Acarol. 15, 21–30. Need, J.T., Dale, W.E., Keirans, J.E., Dasch, G.A., 1991. Annotated list of ticks (Acari: Ixodidae, Argasidae) reported in Peru: distribution, hosts, and bibliography. J. Med. Entomol. 28, 590–597. Ogrzewalska, M., Pacheco, R.C., Uezu, A., Richtzenhain, L.J., Ferreira, F., Labruna, M.B., 2009. Rickettsial infection in Amblyomma nodosum ticks (Acari: Ixodidae) from Brazil. Ann. Trop. Med. Parasitol. 103, 413–425. Ogrzewalska, M., Uezu, A., Labruna, M.B., 2011. Ticks (Acari: Ixodidae) infesting wild birds in the Atlantic Forest in northeastern Brazil, with notes on rickettsial infection in ticks. Parasitol. Res. 108, 665–670. Pacheco, R.C., Rosa, S., Richtzenhain, L.J., Szabó, M.P.J., Labruna, M.B., 2008. Isolation of Rickettsia bellii from Amblyomma ovale and Amblyomma incisum ticks from southern Brazil. Rev. MVZ Cordoba 13, 1273–1279. Pinter, A., Labruna, M.B., 2006. Isolation of Rickettsia rickettsii and Rickettsia bellii in cell culture from the tick Amblyomma aureolatum in Brazil. Ann. N. Y. Acad. Sci. 1078, 523–529. Raoult, D., Roux, V., 1997. Rickettsioses as paradigms of new or emerging infectious diseases. Clin. Med. Rev. 10, 694–719. Schulenberg, T.S., Stotz, D.F., Lane, D.F., O’Neil, J.P., Parker III, A.P., 2010. Birds of Peru, revised and updated edition. Princeton University Press, Princeton and Oxford, p. 664. ˜ P., Ceballos, L.A., Gürtler, R.E., Kitron, U., Farber, M., 2010. Tomassone, L., Nunez, Detection of “Candidatus Rickettsia sp. strain Argentina” and Rickettsia bellii in Amblyomma ticks (Acari: Ixodidae) from northern Argentina. Exp. Appl. Acarol. 52, 93–100.
Contents lists available at SciVerse ScienceDirect
Ticks and Tick-borne Diseases journal homepage: www.elsevier.de/ttbdis
Short communication
Rickettsia bellii in ticks Amblyomma varium Koch, 1844, from birds in Peru Maria Ogrzewalska a, Ivan Literak b,c,∗, Jorge M. Cardenas-Callirgos d, Miroslav Capek e, Marcelo B. Labruna a a
Departamento de Medicina Veterinaria Preventiva e Saude Animal, Faculdade de Medicina Veterinaria e Zootecnia, Universidade de Sao Paulo, Sao Paulo, Brazil Department of Biology and Wildlife Diseases, Faculty of Veterinary Hygiene and Ecology, University of Veterinary and Pharmaceutical Sciences Brno, Brno, Czech Republic CEITEC VFU, University of Veterinary and Pharmaceutical Sciences Brno, Brno, Czech Republic d Laboratorio de Invertebrados, Museo de Historia Natural, Facultad de Ciencias Biologicas, Universidad Ricardo Palma, Lima, Peru e Institute of Vertebrate Biology, Academy of Sciences of the Czech Republic, v. v. i., Brno, Czech Republic b c
a r t i c l e
i n f o
Article history: Received 30 March 2012 Received in revised form 11 May 2012 Accepted 23 May 2012 Keywords: Rickettsia bellii Ticks Amblyomma calcaratum Birds Peru
a b s t r a c t Amazonian birds were caught and examined for the presence of ectoparasites in the Allpahuayo Mishana National Reserve near Iquitos, Peru, from 13 to 16 August 2011. A total of 40 birds representing 16 species were examined. Two birds (5%) were infested with 2 larvae of Amblyomma varium Koch, 1844, and one nymph of A. calcaratum Neumann, 1899. The 2 larvae of A. varium were infected with Rickettsia bellii. This is the first report of R. bellii in A. varium and also the first record of this rickettsia in Peru. In addition, an immature A. calcaratum is reported from Peru for the first time. © 2012 Elsevier GmbH. All rights reserved.
Introduction
Materials and methods
Information on the tick fauna and tick-borne pathogens in Peru is scarce. The presence of 50 tick species (35 of the family Ixodidae, 15 of Argasidae) has been confirmed in Peru (Need et al., 1991; Caceres et al., 2002; Mendoza-Uribe and Chaves-Chorocco, 2004; Blair et al., 2004; Guglielmone et al., 2005; Labruna et al., 2005, 2010; Nava et al., 2010). Wild birds are commonly parasitized by larvae and nymphs of ticks, and they are undeniably among tick hosts with the highest mobility, and therefore they may be regarded as hosts with the highest potential for dispersal of ticks and tick-borne diseases (Hoogstraal, 1961). Bacteria of the genus Rickettsia are obligate intracellular organisms that infect invertebrate hosts worldwide. Some of them cause disease (i.e., rickettsioses) in warm-blooded animals and humans, to whom they are transmitted by hematophagous vectors, mostly ticks (Raoult and Roux, 1997). The present study notes tick infestations on wild birds in the Amazonian forest of Peru and rickettsial infection proved in these ticks.
Mist nets were used to trap wild birds in Peru in summer 2011. Birds trapped were subjected to examination for the presence of ectoparasites including ticks. The ticks were collected after naked eye examination using a tweezer and preserved in 96% ethyl alcohol. After examination, all the birds were released back into the wild. Four study sites were as follows: (i) Refugio de Vida Silvestre Los Pantanos de Villa, Lima, 12◦ 13 S, 76◦ 59W, at sea level (a.s.l.), 50 birds of 10 species were examined from 19 to 20 July 2011. (ii) Centro URKU, Tarapoto, 06◦ 27 S, 76◦ 21 W, 410 m a.s.l., 39 birds of 21 species were examined from 8 to 10 August 2011. (iii) Reserva Nacional Allpahuayo Mishana, Iquitos, 03◦ 58 S, 73◦ 25 W, 134 m a.s.l., 40 birds of 16 species were examined from 13 to 16 August 2011. (iv) Cascay, Huanuco, 09◦ 50 S, 76◦ 08 W, 1845 m a.s.l., 104 birds of 18 species were examined from 20 to 23 August 2011. A total of 233 birds of 57 species were examined (Table 1). The birds were identified using the field guide by Schulenberg et al. (2010). Nymphal ticks were identified according to Martins et al. (2010). To identify larvae, we used molecular tools as described elsewhere (Ogrzewalska et al., 2011). Larvae collected were individually submitted to DNA extraction and then to PCR using primers targeting the tick mitochondrial 16S rDNA gene (Mangold et al., 1998) and the 12S rDNA gene (Beati and Keirans, 2001). Amplified products were purified and DNA-sequenced as previously described (Labruna et al., 2004a) and compared with NCBI Nucleotide BLAST searches (Altschul et al., 1990).
∗ Corresponding author at: Department of Biology and Wildlife Diseases, Faculty of Veterinary Hygiene and Ecology, University of Veterinary and Pharmaceutical Sciences Brno, Palackeho 1-3, 612 42 Brno, Czech Republic. E-mail address: [email protected] (I. Literak). 1877-959X/$ – see front matter © 2012 Elsevier GmbH. All rights reserved. http://dx.doi.org/10.1016/j.ttbdis.2012.05.003
M. Ogrzewalska et al. / Ticks and Tick-borne Diseases 3 (2012) 254–256 Table 1 A list of birds examined (PV, Refugio de Vida Silvestre Los Pantanos de Villa, Lima; TR, Centro URKU, Tarapoto; IQ, Reserva Nacional Allpahuayo Mishana, Iquitos; HU, Cascay, Huanuco). Family/species Columbidae Columbina cruziana Columbina minuta Leptotila rufaxilla Leptotila verreauxi Trochilidae Amazilia amazilia Amazilia fimbriata Amazilia chionogaster Amazilia lactea Doryfera johannae Glaucis hirsutus Phaethornis atrimentalis Phaethornis superciliosus Thalurania furcata Threnetes leucurus Cuculidae Crotophaga ani Ardeidae Butorides striata Ixobrychus exilis Laridae Larus belcheri Bucconidae Bucco macrodactylus Falconidae Falco sparverius Thamnophilidae Thamnophilus schistaceus Furnariidae Dendrocincla fuliginosa Glyphorhynchus spirurus Phleocryptes melanops Pipridae Lepidothrix coronata Machaeropterus pyrocephalus Manacus manacus Pipra fasciicauda Cotingidae Pachyramphus polychopterus Tyrannidae Elaenia spectabilis Leptopogon amaurocephalus Mionectes oleagineus Pitangus lictor Pitangus sulphuratus Sayornis nigricans Tachuris rubrigastra Todirostrum cinereum Tolmomyias flaviventris Tyrannus melancholicus Vireonidae Vireo olivaceus Hirundinidae Pygochelidon cyanoleuca Troglodytidae Campylorhynchus fasciatus Troglodytes aedon Polioptilidae Ramphocaenus melanurus Turdidae Turdus ignobilis Fringillidae Carduellis magellanica Emberizidae Zonotrichia capensis Cardinalidae Pheucticus chrysogaster Thraupidae Diglossa sittoides Oryzoborus angolensis Ramphocelus carbo Sicalis luteola
No. of birds examined 2 7 3 1 1 1 19 2 1 5 4 10 1 3
Locations HU HU IQ HU PV IQ HU TR TR TR (2), IQ (3) TR TR (5), IQ (5) IQ TR
4
HU
2 2
PV PV
1
PV
1
TR
2
PV (1), HU (1)
2
TR
3 5 9
IQ IQ PV
4 3 1 2
IQ TR (2), IQ (1) IQ TR
4
TR (2), IQ (2)
1 1 7 1 1 3 23 1 1 2
TR TR TR (1), IQ (6) IQ TR HU PV HU TR HU
2
TR
2
HU
6 7
HU PV
1
TR
2
TR (1), HU (1)
255
Table 1 (Continued) Family/species
No. of birds examined
Locations
Sporophila nigricollis Sporophila telasco Tachyphonus luctuosus Thraupis episcopus Tiaris obscurus Total
9 1 1 19 3 233
HU PV TR HU TR
Two larvae were individually tested for the presence of Rickettsia by PCR targeting a fragment of the gltA gene, which occurs in all Rickettsia species (Labruna et al., 2004a), and another PCR targeting a fragment of the ompA gene that occurs only in Rickettsia species of the spotted fever group (SFG). PCR products were DNA-sequenced and submitted to BLAST analysis to determine similarities to other Rickettsia species. Results The ticks were found only on Amazonian birds in the Reserva Nacional Allpahuayo Mishana where 40 birds belonging to 16 species were captured. Two birds (5%) were infested with ticks: one of 3 grey-fronted doves Leptotila rufaxilla (Columbiformes: Columbidae), and one of 2 bicolored antbirds Gymnopithys leucaspis (Passeriformes: Thamnophilidae). Two larvae of Amblyomma sp. were found on L. rufaxilla. Their 16S rDNA sequences were identical to each other, and 88.6% (364/411 bp) similar to A. brasiliense (FJ424399) and 87.6% (361/412 bp) to A. maculatum (L34318). Their 12S rDNA sequences were identical to each other and 97.6% (331/339 bp) similar to A. varium (AY974339) from Brazil and 99.4% (313/315 bp) to A. varium (AY342283) from Peru. As there is no specimen of A. varium available in GenBank, the found larvae were identified as A. varium based on the high similarity of the sequences of the 12S rDNA fragment to that species. Based on the gltA gene PCR, these 2 A. varium larvae were found infected with a Rickettsia 99.1% (333/336) similar to R. bellii strain Pontal, previously isolated from A. nodosum in Brazil (EU567181), and R. bellii strain HJ7 (DQ865204) isolated from Haemaphysalis juxtakochi Cooley, 1946, also in Brazil. This high similarity (>99%) of the gltA partial sequences is indicative of that these 2 Amblyomma larvae from Peru were infected by R. bellii. The samples were negative to PCR targeting the ompA gene, which was expected because R. bellii is not an SFG species. One nymph of A. calcaratum Neumann, 1899, was found on G. leucaspis. The nymph was not tested for the presence of rickettsia, and it was deposited at the tick collection “Colecao Nacional de Carrapatos” (CNC) of the University of Sao Paulo, Brazil (accession number 2155). GenBank nucleotide sequence accession numbers for the partial sequences generated in the present study are JQ771810 for the R. bellii partial gltA sequence, and JQ771811 and JQ771812 for the mitochondrial 12S rDNA and 16S rDNA gene of the A. varium larva, respectively. Discussion
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A. calcaratum has been reported in various countries of Central and South America (Guglielmone et al., 2003). However, this is the second report of A. calcaratum in Peru and the first report of an immature stage of A. calcaratum on a bird in this country. The adult stage parasitizes chiefly anteaters, Tamandua spp. and Myrmecophaga tridactyla, whilst immatures feed mostly on wild passerine birds (Jones et al., 1972; Ogrzewalska et al., 2011). Until now, the known presence of A. calcaratum in Peru has been restricted to 12 adult specimens collected from the southern tamandua Tamandua tetradactyla in the Caserio Menor de Nunhan Jalca, Fundo Santa
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Rosa, Bagua Grande, Utcubamba Amazonas (Caceres et al., 2002). Here, we add one new host species for A. calcaratum, G. leucaspis, which has not previously been reported to be parasitized by this tick species. A. varium is a neotropical tick, known as the sloth’s giant tick, with records from southern Central America to Argentina, including Peru (Guglielmone et al., 2003). Adults are found almost exclusively on mammals of the families Bradypodidae and Magalonychidae (Xenarthra) (Guglielmone et al., 2003; Need et al., 1991), however, the hosts of the immature stages have been still unknown, so we speculate that their hosts could be birds. In this study, both A. varium larvae were infected with R. bellii. R. bellii has been notified as a common Rickettsia in various species of hard ticks from Brazil (Labruna et al., 2004a, b, 2007a; Pinter and Labruna, 2006; Ogrzewalska et al., 2009; Pacheco et al., 2008) and Argentina (Labruna et al., 2007b; Tomassone et al., 2010); however, there is no evidence that it is pathogenic for humans or animals (Labruna, 2009; Labruna et al., 2011). This is the first report of R. bellii in Peru and the first finding of this rickettsia infecting A. varium. Acknowledgments We thank Martina Literakova, Zuzana Literakova, and Blgo. Kember Mejia Carhuanca (Director Programa de Investigacion en Biodiversidad Amazonica, Instituto de Investigaciones de la Amazonia Peruana, Iquitos, Peru) for their cooperation in the field. The field work was covered by a permission from responsible Peruvian authorities licensed for Lic. Mercedes Gonzales de la Cruz (Museo de Historia Natural, Facultad de Ciencias Biologicas, Universidad Ricardo Palma, Lima, Peru). The study was supported in part by the Grant Agency of the Academy of Sciences of the Czech Republic (grant IAA601690901) and by the Ministry of Education, Youth and Sports of the Czech Republic (grant LC06073). Ivan Literak was supported by the project ‘CEITEC – Central European Institute of Technology’ (CZ.1.05/1.1.00/02.0068) of the European Regional Development Fund. References Altschul, S.F., Gish, W., Miller, W., Myers, E.W., Lipman, D.J., 1990. Basic local alignment search tool. J. Mol. Biol. 215, 403–410. Beati, L., Keirans, J.E., 2001. Analysis of the systematic relationships among ticks of the genera Rhipicephalus and Boophilus (Acari: Ixodidae) based on mitochondrial 12S ribosomal DNA gene sequences and morphological characters. J. Parasitol. 87, 32–48. Blair, P.J., Jiang, J., Schoeler, G.B., Moron, C., Anaya, E., Cespedes, M., Cruz, C., Felices, V., Guevara, C., Mendoza, L., Villaseca, P., Sumner, J.W., Richards, A.L., Olson, J.G., 2004. Characterization of spotted fever group rickettsiae in flea and tick specimens from northern Peru. J. Clin. Microbiol. 42, 4961–4967. Caceres, A.G., Beati, L., Keirans, J.E., 2002. First evidence of the occurrence of Amblyomma calcaratum Neumann, 1899 in Peru. Rev. Peru Biol. 9, 116–117. ˜ A., Keirans, J.E., Robbins, R.G., 2003. Ticks (Acari: Guglielmone, A.A., Estrada-Pena, Ixodida) of the Neotropical Zoogeographic Region. International Consortium on Ticks and Tick-borne Diseases, Atlanta, Houten, The Netherlands, p. 173. Guglielmone, A.A., Romero, J., Venzal, J.M., Nava, S., Mangold, A.J., Villavicencio, J., 2005. First record of Haemaphysalis juxtakochi Cooley, 1946 (Acari: Ixodidae) from Peru. Syst. Appl. Acarol. 10, 33–35.
Hoogstraal, H., 1961. Migrating birds and their ectoparasites in relation to disease. East Afr. Med. J. 38, 221–226. Jones, E.K., Clifford, C.M., Keirans, J.E., Kohls, G.M., 1972. The ticks of Venezuela (Acarina: Ixodoidea) with a key to the species of Amblyomma in the western hemisphere. Brigham Young Univ. Sci. Bull. Biol. Ser. 17, 1–40. Labruna, M.B., Whitworth, T., Horta, M.C., Bouyer, D.H., McBride, J.W., Pinter, A., Popov, V., Gennari, S.M., Walker, D.H., 2004a. Rickettsia species infecting Amblyomma cooperi ticks from an area in the state of Sao Paulo, Brazil, where Brazilian spotted fever is endemic. J. Clin. Microbiol. 42, 90–98. Labruna, M.B., Whitworth, T., Bouyer, D.H., McBride, J.W., Camargo, L.M.A., Camargo, E.P., Popov, V., Walker, D.H., 2004b. Rickettsia bellii and Rickettsia amblyommii in Amblyomma ticks from the state of Rondônia, Western Amazon, Brazil. J. Med. Entomol. 41, 1073–1081. Labruna, M.B., Keirans, J.E., Camargo, L.M.A., Ribeiro, A.F., Soares, R.M., Camargo, E.P., 2005. Amblyomma latepunctatum, a valid tick species (Acari: Ixodidae) long misidentified with both Amblyomma incisum and Amblyomma scalpturatum. J. Parasitol. 91, 527–541. Labruna, M.B., Pacheco, R.C., Richtzenhain, L.J., Szabo, M.P., 2007a. Isolation of Rickettsia rhipicephali and Rickettsia bellii from ticks Haemaphysalis juxtakochi in the state of Sao Paulo, Brazil. Appl. Environ. Microbiol. 73, 869–873. Labruna, M.B., Pacheco, R.C., Nava, S., Brandao, P.E., Richtzenhain, L.J., Guglielmone, A.A., 2007b. Infection by Rickettsia bellii and Candidatus “Rickettsia amblyommii” in Amblyomma neumanni ticks from Argentina. Microb. Ecol. 54, 126–133. Labruna, M.B., 2009. Ecology of Rickettsia in South America. Ann. N. Y. Acad. Sci. 1166, 156–166. Labruna, M.B., Romero, M., Martins, T.F., Tobler, M., Ferreira, F., 2010. Ticks of the genus Amblyomma (Acari: Ixodidae) infesting tapirs (Tapirus terrestris) and peccaries (Tayassu pecari) in Peru. Syst. Appl. Acarol. 15, 109–112. Labruna, M.B., Mattar, S., Nava, S., Bermudez, S., Venzal, J.M., Dolz, G., Abarca, K., Romero, L., de Sousa, R., Oteo, J., Zavala-Castro, J., 2011. Rickettsioses in Latin America, Caribbean, Spain and Portugal. Rev. MVZ Córdoba 16, 2435–2457. Mangold, A.J., Bargues, M.D., Mas-Coma, S., 1998. Mitochondrial 16S rDNA sequences and phylogenetic relationships of species of Rhipicephalus and other tick genera among Metastriata (Acari: Ixodidae). Parasitol. Res. 84, 478–484. Martins, T.F., Onofrio, V.C., Barros-Battesti, D.M., Labruna, M.B., 2010. Nymphs of the genus Amblyomma (Acari: Ixodidae) of Brazil: descriptions, redescriptions, and identification key. Ticks Tick-Borne Dis. 1, 75–99. Mendoza-Uribe, L., Chaves-Chorocco, J., 2004. Amplicacion geografica de siete especies de Amblyomma (Acari: Ixodidae) y primer reporte de A. oblongoguttatum Koch, 1844 para Peru. Rev. Per. Ent. 44, 69–72. Nava, S., Velazco, P.M., Guglielmone, A.A., 2010. The first record of Amblyomma longirostre (Koch, 1844) (Acari: Ixodidae) from Peru, with a revision of its host range. Syst. Appl. Acarol. 15, 21–30. Need, J.T., Dale, W.E., Keirans, J.E., Dasch, G.A., 1991. Annotated list of ticks (Acari: Ixodidae, Argasidae) reported in Peru: distribution, hosts, and bibliography. J. Med. Entomol. 28, 590–597. Ogrzewalska, M., Pacheco, R.C., Uezu, A., Richtzenhain, L.J., Ferreira, F., Labruna, M.B., 2009. Rickettsial infection in Amblyomma nodosum ticks (Acari: Ixodidae) from Brazil. Ann. Trop. Med. Parasitol. 103, 413–425. Ogrzewalska, M., Uezu, A., Labruna, M.B., 2011. Ticks (Acari: Ixodidae) infesting wild birds in the Atlantic Forest in northeastern Brazil, with notes on rickettsial infection in ticks. Parasitol. Res. 108, 665–670. Pacheco, R.C., Rosa, S., Richtzenhain, L.J., Szabó, M.P.J., Labruna, M.B., 2008. Isolation of Rickettsia bellii from Amblyomma ovale and Amblyomma incisum ticks from southern Brazil. Rev. MVZ Cordoba 13, 1273–1279. Pinter, A., Labruna, M.B., 2006. Isolation of Rickettsia rickettsii and Rickettsia bellii in cell culture from the tick Amblyomma aureolatum in Brazil. Ann. N. Y. Acad. Sci. 1078, 523–529. Raoult, D., Roux, V., 1997. Rickettsioses as paradigms of new or emerging infectious diseases. Clin. Med. Rev. 10, 694–719. Schulenberg, T.S., Stotz, D.F., Lane, D.F., O’Neil, J.P., Parker III, A.P., 2010. Birds of Peru, revised and updated edition. Princeton University Press, Princeton and Oxford, p. 664. ˜ P., Ceballos, L.A., Gürtler, R.E., Kitron, U., Farber, M., 2010. Tomassone, L., Nunez, Detection of “Candidatus Rickettsia sp. strain Argentina” and Rickettsia bellii in Amblyomma ticks (Acari: Ixodidae) from northern Argentina. Exp. Appl. Acarol. 52, 93–100.