• View in gallery

    Phylogenetic position of “Candidatus Rickettsia goldwasserii.” Phylogenetic position was inferred using the Neighbor-Joining method. The optimal tree with the sum of branch length = 0.78594827 is shown. The percentage of replicate trees in which the associated taxa clustered together in the bootstrap test (1,000 replicates) are shown next to the branches; only bootstrap values of ≥ 80 are shown. The tree is drawn to scale, with branch lengths in the same units as those of the evolutionary distances used to infer the phylogenetic tree. The evolutionary distances were computed using the Kimura 2-parameter method as base substitutions per site. All positions containing gaps, missing data and primer sequences were eliminated from the dataset. Fragments of the four genes were concatenated (gltA-ompA-sca4-ompB), and a total of 2,028 positions were analyzed. Phylogenetic analyses were conducted with MEGA4.21

  • 1.

    Aharonowitz G, Koton S, Segal S, Anis E, Green MS, 1999. Epidemiological characteristics of spotted fever in Israel over 26 years. Clin Infect Dis 29: 13211322.

    • Search Google Scholar
    • Export Citation
  • 2.

    Mumcuoglu KY, Keysary A, Gilead L, 2002. Mediterranean spotted fever in Israel: a tick-borne disease. Isr Med Assoc J 4: 4449.

  • 3.

    Guberman D, Mumcuoglu KY, Keysary A, Ioffe-Uspensky I, Miller J, Galun R, 1996. Prevalence of spotted fever group rickettsiae in ticks from southern Israel. J Med Entomol 33: 979982.

    • Search Google Scholar
    • Export Citation
  • 4.

    Mumcuoglu KY, Ioffe-Uspensky I, Alkrinawi S, Sarov B, Manor E, Galun R, 2001. Prevalence of vectors of the spotted fever group Rickettsiae and murine typhus in a Bedouin town in Israel. J Med Entomol 38: 458461.

    • Search Google Scholar
    • Export Citation
  • 5.

    Harrus S, Lior Y, Ephros M, Grisaru-Soen G, Keysary A, Strenger C, Jongejan F, Waner T, Baneth G, 2007. Rickettsia conorii in humans and dogs: a seroepidemiologic survey of two rural villages in Israel. Am J Trop Med Hyg 77: 133135.

    • Search Google Scholar
    • Export Citation
  • 6.

    Harrus S, Perlman-Avrahami A, Mumcuoglu KY, Morick D, Baneth G, 2011. Molecular detection of Rickettsia massiliae, Rickettsia sibirica mongolitimonae and Rickettsia conorii israelensis in ticks from Israel. Clin Microbiol Infect 17: 176180.

    • Search Google Scholar
    • Export Citation
  • 7.

    Cwilich R, Hadani A, 1969. A note on some cases of human infestation with “hard” ticks (Ixodoidea, Ixodidae). Refu Vet 26: 7981.

  • 8.

    Feldman-Muhsam B, 1986. Ixodid tick attacks on man in Israel: medical implications. Isr J Med Sci 22: 1923.

  • 9.

    Feldman-Muhsam B, Shechter R, 1970. Some notes on the genus Boophilus (Ixodidae), with special reference to species found in Israel. J Med Entomol 7: 677686.

    • Search Google Scholar
    • Export Citation
  • 10.

    Theodor O, Costa M, 1967. Part 1. Ectoparasites. A Survey of the Parasites of Wild Mammals and Birds in Israel. Jerusalem: The Israel Academy of Sciences and Humanities, 117.

    • Search Google Scholar
    • Export Citation
  • 11.

    Feldman-Muhsam B, 1951. A note on East Mediterranean species of the Haemophysalis. Bull Res Counc Isr 1: 96107.

  • 12.

    Waner T, Baneth G, Strenger C, Keysary A, King R, Harrus S, 1999. Antibodies reactive with Ehrlichia canis, Ehrlichia phagocytophila genogroup antigens and the spotted fever group rickettsial antigens, in free-ranging jackals (Canis aureus syriacus) from Israel. Vet Parasitol 82: 121128.

    • Search Google Scholar
    • Export Citation
  • 13.

    Wallach AD, Shanas U, Mumcuoglu KY, Inbar M, 2008. Ectoparasites on reintroduced roe deer Capreolus capreolus in Israel. J Wildl Dis 44: 693696.

    • Search Google Scholar
    • Export Citation
  • 14.

    Pegram R, Clifford C, Walker J, Keirans J, 1987. Clarification of the Rhipicephalus sanguineus group (Acari, Ixodoidea, Ixodidae). I. R. sulcatus Neumann, 1908 and R. turanicus Pomerantsev, 1936. Syst Parasitol 10: 326.

    • Search Google Scholar
    • Export Citation
  • 15.

    Walker J, Keirans J, Horak I, 2000. The genus Rhipicephalus (Acari, Ixoidae). A Guide to the Brown Ticks of the World. Cambridge: Cambridge University Press.

    • Search Google Scholar
    • Export Citation
  • 16.

    Feldman-Muhsam B, 1954. Revision of the genus Hyalomma. Bull Res Counc Isr 64: 150170.

  • 17.

    Leitner M, Yitzhaki S, Rzotkiewicz S, Keysary A, 2002. Polymerase chain reaction-based diagnosis of Mediterranean spotted fever in serum and tissue samples. Am J Trop Med Hyg 67: 166169.

    • Search Google Scholar
    • Export Citation
  • 18.

    Eremeeva ME, Bosserman EA, Demma LJ, Zambrano ML, Blau DM, Dasch GA, 2006. Isolation and identification of Rickettsia massiliae from Rhipicephalus sanguineus ticks collected in Arizona. Appl Environ Microbiol 72: 55695577.

    • Search Google Scholar
    • Export Citation
  • 19.

    Sekeyova Z, Roux V, Raoult D, 2001. Phylogeny of Rickettsia spp. inferred by comparing sequences of ‘gene D', which encodes an intracytoplasmic protein. Int J Syst Evol Microbiol 51: 13531360.

    • Search Google Scholar
    • Export Citation
  • 20.

    Roux V, Raoult D, 2000. Phylogenetic analysis of members of the genus Rickettsia using the gene encoding the outer-membrane protein rOmpB (ompB). Int J Syst Evol Microbiol 50: 14491455.

    • Search Google Scholar
    • Export Citation
  • 21.

    Tamura K, Dudley J, Nei M, Kumar S, 2007. MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol 24: 15961599.

    • Search Google Scholar
    • Export Citation
  • 22.

    Eremeeva ME, Stromdahl EY, 2011. Short report: new spotted fever group Rickettsia in a Rhipicephalus turanicus tick removed from a child in eastern Sicily, Italy. Am J Trop Med Hyg 84: 99101.

    • Search Google Scholar
    • Export Citation
  • 23.

    Sarih M, Socolovschi C, Boudebouch N, Hassar M, Raoult D, Parola P, 2008. Spotted fever group rickettsiae in ticks, Morocco. Emerg Infect Dis 14: 10671073.

    • Search Google Scholar
    • Export Citation
  • 24.

    Raoult D, Fournier PE, Abboud P, Caron F, 2002. First documented human Rickettsia aeschlimannii infection. Emerg Infect Dis 8: 748749.

  • 25.

    Parola P, 2006. Rickettsioses in sub-Saharan Africa. Ann N Y Acad Sci 1078: 4247.

  • 26.

    Shpynov S, Fournier PE, Rudakov N, Tankibaev M, Tarasevich I, Raoult D, 2004. Detection of a Rickettsia closely related to Rickettsia aeschlimannii, “Rickettsia heilongjiangensis,” Rickettsia sp. strain RpA4, and Ehrlichia muris in ticks collected in Russia and Kazakhstan. J Clin Microbiol 42: 22212223.

    • Search Google Scholar
    • Export Citation
  • 27.

    Portillo A, Santibanez P, Santibanez S, Perez-Martinez L, Oteo JA, 2008. Detection of Rickettsia spp. in Haemaphysalis ticks collected in La Rioja, Spain. Vector Borne Zoonotic Dis 8: 653658.

    • Search Google Scholar
    • Export Citation
  • 28.

    Mura A, Socolovschi C, Ginesta J, Lafrance B, Magnan S, Rolain JM, Davoust B, Raoult D, Parola P, 2008. Molecular detection of spotted fever group rickettsiae in ticks from Ethiopia and Chad. Trans R Soc Trop Med Hyg 102: 945949.

    • Search Google Scholar
    • Export Citation
  • 29.

    Fernandez-Soto P, Diaz Martin V, Perez-Sanchez R, Encinas-Grandes A, 2009. Increased prevalence of Rickettsia aeschlimannii in Castilla y Leon, Spain. Eur J Clin Microbiol Infect Dis 28: 693695.

    • Search Google Scholar
    • Export Citation
  • 30.

    Bitam I, Parola P, Matsumoto K, Rolain JM, Baziz B, Boubidi SC, Harrat Z, Belkaid M, Raoult D, 2006. First molecular detection of R. conorii, R. aeschlimannii, and R. massiliae in ticks from Algeria. Ann N Y Acad Sci 1078: 368372.

    • Search Google Scholar
    • Export Citation
  • 31.

    Loftis AD, Reeves WK, Szumlas DE, Abbassy MM, Helmy IM, Moriarity JR, Dasch GA, 2006. Rickettsial agents in Egyptian ticks collected from domestic animals. Exp Appl Acarol 40: 6781.

    • Search Google Scholar
    • Export Citation
  • 32.

    Psaroulaki A, Ragiadakou D, Kouris G, Papadopoulos B, Chaniotis B, Tselentis Y, 2006. Ticks, tick-borne rickettsiae, and Coxiella burnetii in the Greek Island of Cephalonia. Ann N Y Acad Sci 1078: 389399.

    • Search Google Scholar
    • Export Citation
  • 33.

    Fernandez-Soto P, Encinas-Grandes A, Perez-Sanchez R, 2003. Rickettsia aeschlimannii in Spain: molecular evidence in Hyalomma marginatum and five other tick species that feed on humans. Emerg Infect Dis 9: 889890.

    • Search Google Scholar
    • Export Citation
  • 34.

    Pretorius AM, Birtles RJ, 2002. Rickettsia aeschlimannii: a new pathogenic spotted fever group Rickettsia, South Africa. Emerg Infect Dis 8: 874.

    • Search Google Scholar
    • Export Citation
  • 35.

    Brouqui P, Parola P, Fournier PE, Raoult D, 2007. Spotted fever rickettsioses in southern and eastern Europe. FEMS Immunol Med Microbiol 49: 212.

    • Search Google Scholar
    • Export Citation
  • 36.

    Letaief A, 2006. Epidemiology of rickettsioses in North Africa. Ann N Y Acad Sci 1078: 3441.

  • 37.

    Parola P, Socolovschi C, Jeanjean L, Bitam I, Fournier PE, Sotto A, Labauge P, Raoult D, 2008. Warmer weather linked to tick attack and emergence of severe rickettsioses. PLoS Negl Trop Dis 2: e338.

    • Search Google Scholar
    • Export Citation
  • 38.

    Garcia-Garcia JC, Portillo A, Nunez MJ, Santibanez S, Castro B, Oteo JA, 2010. A patient from Argentina infected with Rickettsia massiliae. Am J Trop Med Hyg 82: 691692.

    • Search Google Scholar
    • Export Citation
  • 39.

    Vitale G, Mansuelo S, Rolain JM, Raoult D, 2006. Rickettsia massiliae human isolation. Emerg Infect Dis 12: 174175.

  • 40.

    Rolain JM, Maurin M, Vestris G, Raoult D, 1998. In vitro susceptibilities of 27 rickettsiae to 13 antimicrobials. Antimicrob Agents Chemother 42: 15371541.

    • Search Google Scholar
    • Export Citation
  • 41.

    Raoult D, Fournier PE, Eremeeva M, Graves S, Kelly PJ, Oteo JA, Sekeyova Z, Tamura A, Tarasevich I, Zhang L, 2005. Naming of Rickettsiae and rickettsial diseases. Ann N Y Acad Sci 1063: 112.

    • Search Google Scholar
    • Export Citation
  • 42.

    Murray RG, Stackebrandt E, 1995. Taxonomic note: implementation of the provisional status Candidatus for incompletely described procaryotes. Int J Syst Bacteriol 45: 186187.

    • Search Google Scholar
    • Export Citation
  • 43.

    Goldwasser RA, Steiman Y, Klingberg W, Swartz TA, Klingberg MA, 1974. The isolation of strains of rickettsiae of the spotted fever group in Israel and their differentiation from other members of the group by immunofluorescence methods. Scand J Infect Dis 6: 5362.

    • Search Google Scholar
    • Export Citation
 
 
 
 

 

 

 

 

 

 

 

 

 

Spotted Fever Group Rickettsiae in Ticks Collected from Wild Animals in Israel

View More View Less
  • Israel Institute for Biological Research, Ness Ziona, Israel; Centers for Disease Control and Prevention, Atlanta, Georgia; Department of Microbiology and Molecular Genetics, Hebrew University-Hadassah Medical School, Jerusalem, Israel; University of Haifa, Haifa, Israel; Israel Nature and Parks Authority, Jerusalem

We report molecular evidence for the presence of spotted fever group rickettsiae (SFGR) in ticks collected from roe deer, addax, red foxes, and wild boars in Israel. Rickettsia aeschlimannii was detected in Hyalomma marginatum and Hyalomma detritum while Rickettsia massiliae was present in Rhipicephalus turanicus ticks. Furthermore, a novel uncultured SFGR was detected in Haemaphysalis adleri and Haemaphysalis parva ticks from golden jackals. The pathogenicity of the novel SFGR for humans is unknown; however, the presence of multiple SFGR agents should be considered when serological surveillance data from Israel are interpreted because of significant antigenic cross-reactivity among Rickettsia. The epidemiology and ecology of SFGR in Israel appear to be more complicated than was previously believed.

Introduction

Tick-borne spotted fever group (SFG) rickettsioses are caused by obligatory intracellular gram-negative bacteria of the genus Rickettsia. In Israel, Mediterranean spotted fever (MSF) caused by Rickettsia conorii subsp. israelensis is considered to be the primary cause of spotted fever group rickettsiosis associated with brown dog ticks, Rhipicephalus sanguineus Latreille.1,2

To gauge the prevalence of SFGR in southern Israel, the hemolymph test showed the presence of rickettsiae in both Rh. sanguineus and Rhipicephalus turanicus collected from agricultural settlements in Israel.3 In the Negev region a correlation among the density of domestic animals, their ectoparasites (Rh. sanguineus, Rh. turanicus, and Hyalomma sp. ticks), and the incidence of spotted fever group rickettsiae was demonstrated.4 Serology was also a sensitive indicator for the presence and magnitude of human and canine exposure to ticks and to SFG rickettsiae (SFGR) based on the prevalence of immunoglobulin G (IgG)-antibodies to Rickettsia conorii in two rural villages in Israel.5 Recently Rickettsia massiliae and Rickettsia sibirica mongolotimonae were found in questing adult ticks collected from the vegetation in different parts of Israel.6 Furthermore, the presence of R. massiliae DNA sequences was detected in a Rh. sanguineus tick picked from the scalp of a pediatric patient in the north of Israel (Keysary A and others, unpublished data).

In Israel ticks such as Rh. sanguineus sensu lato, Rhipicephalus bursa Canestrini and Fanzago, Hyalomma marginatum Koch, and Haemaphysalis sulcata Canestrini and Fanzago have been found attached to humans.7,8 Ixodid ticks, particularly Rh. sanguineus and Rh. turanicus Pomerantsev, are quite common on domestic and wild animals. Rhipicephalus (Boophilus) kohlsi has been found on the hilly land of the Mediterranean phytogeographic area, mainly on goats and sheep, whereas small numbers of this tick were also found on cattle, mules, horses, and camels.9 Adults of H. marginatum and Hyalomma detritum ticks occur on cattle and horses, although their larvae and nymphs can be found on rodents and birds.10 Previously, Haemaphysalis adleri was found on the golden jackal (Canis aureus), red fox (Vulpes vulpes), jungle cat (Felis chauss), and the desert cat (Felis lybica), whereas Haemaphysalis parva was found on dogs, golden jackal, gray wolf (Canis lupus), red fox, and hedgehog (Erinaceus europeus).10,11

In contrast to studies of domestic animals and humans, there is a dearth of literature on the incidence of SFGR in populations of wild animals and their ticks in Israel. Only a single serological study showed high titers of SFG-rickettisal antibodies was detected in a substantial number of free-ranging jackals (Canis aureus syriacus) in Israel.12

We report here molecular evidence for the occurrence of two known human pathogenic species of SFGR, Rickettsia aeschlimannii in Hyalomma ticks, and Rickettsia massiliae in Rh. turanicus ticks collected from wild animals in Israel. In addition, we report the presence of a novel uncultivated SFGR found in Haemaphysalis ticks collected from golden jackals.

Material and Methods

One hundred eighty-one ticks were collected from 6 addax (Addax nasomaculatus), 7 red foxes (V. vulpes), 5 wild boars (Sus scrofa), and 3 golden jackals (Canis aureus); and from 4 roe deer (Capreolus capreolus) (descendant of European animals that were reintroduced into Israel in the 1980s and 1990s).13

Tick collections were performed randomly from animals that had been immobilized for different reasons, live-trapped or found dead. Fallow deer were sampled at Hai Bar Carmel in the north of Israel, addax at Hai Bar Yotvata in the Arava Rift valley in the south of Israel, and the rest from different sites (Table 1).

Table 1

Prevalence of spotted fever group Rickettsia DNA in ticks picked from different hosts*

Tick speciesHostSite of collectionNumber of PCR-positive ticks/number of ticks testedSequence identification (NCBI GenBank accession no.)
Rhipicephalus turanicusRoe deerCarmel Mountains (northern Israel)22/41R. massiliae (GQ856265, GQ856267)
Red foxEin Chemed Even Yehuda (central Israel)2/17
BoarGranot (northern Israel)1/5
Hyalomma marginatumRoe deerCarmel Mountains (northern Israel)10/14R. aeschlimannii (GQ856266, GQ856268)
Hyalomma dentritumAddaxYotveta Hai-Bar (southern Israel)2/15
Hyalomma sp.BoarEin Yakov (northern Israel)1/4
Red foxKfar Shimon (central Israel)1/2
Haemaphysalis adleriGolden jackalWadi Ara (central Israel)1/5Candidatus Rickettsia goldwasserii” HM136923-HM136926
Haemaphysalis parvaGolden jackalWadi Ara (central Israel)1/8Candidatus Rickettsia goldwasserii” HM136927-HM136930
Rhipicephalus sanguineusRoe deerCarmel Mountains (northern Israel)0/40Not applicable
Rhipicephalus (Boophilus) kohlsiRoe deerCarmel Mountains (northern Israel)0/14
Haemaphysalis adleriRed foxBeth-Shemesh (central Israel)0/6
BoarGolan Heights (northern Israel)0/2
Haemaphysalis parvaRed foxBeth-Shemesh (central Israel)0/2
BoarGolan Heights (northern Israel)0/6

PCR = polymerase chain reaction.

Ticks were identified to species using standard taxonomic keys9,11,1416 and comprised 40 Rh. sanguineus, 63 Rh. turanicus, 14 Rhipicephalus (Boophilus) kohlsi Hoogstraal and Kaiser, 14 Hyalomma marginatum Koch, 15 Hyalomma dentritum, 13 Haemaphysalis adleri Feldman-Muhsam, and 16 Haemaphysalis parva Neumann (Table 1). The six specimens of Hyalomma ticks were identified only to the genus.

The ticks were kept in 70% ethanol. DNA was extracted using the QIAamp Minikit (QIAGEN Inc., Valencia, CA), according to the manufacturer's instructions.

Tick extracts were first tested for rickettsial DNA by nested polymerase chain reaction (PCR) to amplify a fragment of 17 kDa protein antigen gene followed by restriction fragment length polymorphism analysis as described previously.17 Species identification of SFG Rickettsia was done by sequencing of 70–602 nucleotide fragments of the outer membrane protein A (OmpA) and 17 kDa protein gene fragments as described previously.18 Multiple locus sequence analysis included amplification and sequencing fragments of gltA, ompA, ompB, and sca4 as described previously.1820 New sequences generated during this study were submitted to NCBI GenBank under the following accession nos.: R. aeschlimannii - GQ856266 and GQ856268, R. massiliae - GQ856265 and GQ856267, and uncultured SFGR - HM136923-HM136930. Phylogenetic analysis was conducted using MEGA4.21

Results

DNA was extracted from 181 ticks collected from 25 wild animals of five species: roe deer, addax, red foxes, golden jackals, and wild boars (Table 1). DNA of SFGR was detected in 41 ticks (22.7% prevalence for the entire study).

DNA of R. massilliae was detected in 25 of 63 Rh. turanicus ticks, collected from roe deer, foxes, and boars. Nucleotide sequences were identical among all ompA amplicons obtained from Rh. turanicus and had 99% nucleotide identity to the homologous ompA fragment of both R. massiliae Mtu5 and Bar29 and 100% similarity to Mtu1 strain.

No ompA sequence differences were observed in DNA from R. aeschlimannii detected in 14 Hyalomma ticks out of 35 collected from roe deer, addax, boars and foxes. DNA of a novel SFGR was found in two Haemaphysalis species ticks: H. adleri (in 1 of 5 ticks) and H. parva (in 1 of 8 ticks) from golden jackals (Table 1). Homologous rickettsial fragments of gltA, ompA, ompB, and sca4 amplified from these two different ticks were found to be identical. The 381 bp gltA fragment sequenced had 99% sequence similarity and at least 1 to 3 unique single-nucleotide polymorphisms (SNP) compared with gltA fragment of Rickettsia honei, Rickettsia africae, Rickettsia slovaca, and Rickettsia japonica. The 575 bp ompA fragment had < 96% sequence similarity to ompA of R. honei and R. slovaca, R. africae and Rickettsia sibirica mongolotimonae, corresponding to 18–21 SNP. The 1,765 bp sca4 fragment had < 97% sequence similarity to the homologous fragments of R. slovaca and other SFGR encompassing multiple SNPs and several unique insertion/deletion (INDEL). The 4,899 bp ompB fragment had ≤ 97% sequence similarity to the nearest rickettsial relative. Phylogenetic analysis of the four concatenated gene fragments, gltA-ompA-sca4-ompB indicated that the nucleotide sequences are those of a SFGR belonging to a novel phylogenetic lineage that appears to be most related to “Candidatus Rickettsia siciliensis” that was recently found in Rh. turanicus (Figure 1).22Candidatus Rickettsia siciliensis” and the SFGR from H. adleri and H. parva ticks share 99%, 95%, 98%, and 96% nucleotide sequence similarity in their homologous fragments of gltA (HM014438), ompA (HM014439), sca4 (HM014440), and ompB (HM014441), respectively.

Figure 1.
Figure 1.

Phylogenetic position of “Candidatus Rickettsia goldwasserii.” Phylogenetic position was inferred using the Neighbor-Joining method. The optimal tree with the sum of branch length = 0.78594827 is shown. The percentage of replicate trees in which the associated taxa clustered together in the bootstrap test (1,000 replicates) are shown next to the branches; only bootstrap values of ≥ 80 are shown. The tree is drawn to scale, with branch lengths in the same units as those of the evolutionary distances used to infer the phylogenetic tree. The evolutionary distances were computed using the Kimura 2-parameter method as base substitutions per site. All positions containing gaps, missing data and primer sequences were eliminated from the dataset. Fragments of the four genes were concatenated (gltA-ompA-sca4-ompB), and a total of 2,028 positions were analyzed. Phylogenetic analyses were conducted with MEGA4.21

Citation: The American Society of Tropical Medicine and Hygiene 85, 5; 10.4269/ajtmh.2011.10-0623

Rickettsial DNA was not detected in Rh. sanguineus and Rh. (B.) kohlsi collected on roe deer or H. adleri from boar and red fox (Table 1).

Discussion

We detected R. aeschlimannii in Hyalomma sp. ticks and R. massiliae in Rh. turanicus ticks collected from wild animals and captive bred wildlife in Israel (Table 1). We also characterized a previously unknown SFGR in H. adleri and H. parva from jackals.

Rickettsia aeschlimannii was first described from H. marginatum marginatum ticks from Morocco in 199723 and was subsequently detected in 2002 in a patient returning from Morocco.24 Subsequently, the presence of R. aeschlimannii has been demonstrated in H. marginatum marginatum ticks with a distribution from Portugal and northern Spain to Kazakhstan and from Mediterranean countries to South Africa.25,26 The prevalence of R. aeschlimannii in H. marginatum ticks tested ranges from 1.8% to 64% in different studies.23,2729 Rickettsia aeschlimannii has also been detected in Hyalomma aegyptum (L.) in Algeria,30 Haemaphysalis inermis Birula in Spain,27 Hyalomma marginatum rufipes Koch in Egypt, Ethiopia, and Chad,28,31 Hyalomma anatolicum excavatum Koch from the Greek Island of Cephalonia,32 and Hyalomma dromedarii Koch and Hyalomma impeltatum Schulze and Schlottke from Egypt.31 Furthermore, five other human biting tick species including Haemaphysalis punctata Canestrini and Fanzago, Ixodes ricinus (L.), Rh. bursa, and Rh. sanguineus collected from Spanish patients were shown to contain DNA of R. aeschlimannii.33 The association of Rhipicephalus complex ticks with R. aeschlimannii is probably not surprising, because it was also detected in Rhipicephalus apendiculatus Neumann ticks collected from a patient suffering from R. aeschlimannii infection.34

Rickettsia massiliae has been detected in Rh. sanguineus, Rhipicephalus sulcatus Neumann, Rhipicephalus lunulatus Neumann, Rhipicephalus muhsamae Morel and Vassiliades, Rhipicephalus senegalensis Koch, Rh. bursa, and Rh. turanicus.23,28,3537 Rickettsia massiliae has been detected in Rhipicephalus ticks in Europe, Africa, and in South and North America.3739 There are several confirmed clinical cases caused by R. massiliae reported in the peer-reviewed literature.3739 Furthermore, it is believed that R. massiliae is responsible for cases of SFG rickettsioses resistant to rifampin in Catalonia, Spain,39 which corresponds closely to the observation that R. massiliae is naturally resistant to this antibiotic.40

The SFGR detected in H. adleri and H. parva has unique genetic characteristics that meet the minimum current requirement for identification as a new species based on the proposed molecular similarity criteria: ≤ 99.9%, ≤ 98.8%, ≤ 99.2%, and ≤ 99.3% for the gltA, ompA and ompB, and sca4, respectively, to its closest SFGR relative.27,41 We cannot propose a formal species description because only two specimens from different tick species were analyzed and a rickettsial isolate was not established to complete its characterization. However, we can assign Candidatus status to this yet uncultivated SFGR Rickettsia and name it “Candidatus Rickettsia goldwasserii” in recognition of Dr. Robert A. Goldwasser for his work on rickettsiae and rickettsial diseases in Israel and his important contributions to the development of indirect fluorescent antibody assays for rickettsiae.42,43 Further detailed study will be necessary to establish the prevalence and distribution of this SFGR in Haemaphysalis ticks from Israel, its primary vector and reservoir, and its ability to cause human and animal rickettsioses.

In Israel, ixodid ticks, which have been found on humans include 13 various species of Hyalomma, the sheep tick, Rh. bursa and H. sulcata as documented by Feldman-Muhsam.8 Cwilich and Hadani found H. excavatum, H. detritum, H. marginatum, Rh. sanguineus, Rh. turanicus, and Rh. bursa on humans.7 To the best of our knowledge, there is no published document showing that H. parva and H. adleri infest humans.

Our results provide evidence for the presence of R. aeschlimannii and confirm the evidence for the presence of R. massiliae in Israel.6 Further surveillance will be needed to characterize tick and animal reservoir for each SFGR reported.

Serological diagnosis of spotted fever infections cannot distinguish between those caused by R. massiliae, R. aeschlimannii, and R. conorii or potentially other SFGR because of the strong cross-reaction among the spotted fever group rickettsiae. Definitive diagnosis of the specific etiological SFGR agent causing rickettsioses in Israel requires molecular techniques conducted on whole blood and skin biopsy samples collected during the acute stage of illness and before antibiotic administration.

ACKNOWLEDGMENTS:

We thank Arianna Salazar for laboratory assistance, rangers of the Israel Nature and Parks Authority for field assistance, and Gregory A. Dasch for reviewing the manuscript and helpful suggestions.

  • 1.

    Aharonowitz G, Koton S, Segal S, Anis E, Green MS, 1999. Epidemiological characteristics of spotted fever in Israel over 26 years. Clin Infect Dis 29: 13211322.

    • Search Google Scholar
    • Export Citation
  • 2.

    Mumcuoglu KY, Keysary A, Gilead L, 2002. Mediterranean spotted fever in Israel: a tick-borne disease. Isr Med Assoc J 4: 4449.

  • 3.

    Guberman D, Mumcuoglu KY, Keysary A, Ioffe-Uspensky I, Miller J, Galun R, 1996. Prevalence of spotted fever group rickettsiae in ticks from southern Israel. J Med Entomol 33: 979982.

    • Search Google Scholar
    • Export Citation
  • 4.

    Mumcuoglu KY, Ioffe-Uspensky I, Alkrinawi S, Sarov B, Manor E, Galun R, 2001. Prevalence of vectors of the spotted fever group Rickettsiae and murine typhus in a Bedouin town in Israel. J Med Entomol 38: 458461.

    • Search Google Scholar
    • Export Citation
  • 5.

    Harrus S, Lior Y, Ephros M, Grisaru-Soen G, Keysary A, Strenger C, Jongejan F, Waner T, Baneth G, 2007. Rickettsia conorii in humans and dogs: a seroepidemiologic survey of two rural villages in Israel. Am J Trop Med Hyg 77: 133135.

    • Search Google Scholar
    • Export Citation
  • 6.

    Harrus S, Perlman-Avrahami A, Mumcuoglu KY, Morick D, Baneth G, 2011. Molecular detection of Rickettsia massiliae, Rickettsia sibirica mongolitimonae and Rickettsia conorii israelensis in ticks from Israel. Clin Microbiol Infect 17: 176180.

    • Search Google Scholar
    • Export Citation
  • 7.

    Cwilich R, Hadani A, 1969. A note on some cases of human infestation with “hard” ticks (Ixodoidea, Ixodidae). Refu Vet 26: 7981.

  • 8.

    Feldman-Muhsam B, 1986. Ixodid tick attacks on man in Israel: medical implications. Isr J Med Sci 22: 1923.

  • 9.

    Feldman-Muhsam B, Shechter R, 1970. Some notes on the genus Boophilus (Ixodidae), with special reference to species found in Israel. J Med Entomol 7: 677686.

    • Search Google Scholar
    • Export Citation
  • 10.

    Theodor O, Costa M, 1967. Part 1. Ectoparasites. A Survey of the Parasites of Wild Mammals and Birds in Israel. Jerusalem: The Israel Academy of Sciences and Humanities, 117.

    • Search Google Scholar
    • Export Citation
  • 11.

    Feldman-Muhsam B, 1951. A note on East Mediterranean species of the Haemophysalis. Bull Res Counc Isr 1: 96107.

  • 12.

    Waner T, Baneth G, Strenger C, Keysary A, King R, Harrus S, 1999. Antibodies reactive with Ehrlichia canis, Ehrlichia phagocytophila genogroup antigens and the spotted fever group rickettsial antigens, in free-ranging jackals (Canis aureus syriacus) from Israel. Vet Parasitol 82: 121128.

    • Search Google Scholar
    • Export Citation
  • 13.

    Wallach AD, Shanas U, Mumcuoglu KY, Inbar M, 2008. Ectoparasites on reintroduced roe deer Capreolus capreolus in Israel. J Wildl Dis 44: 693696.

    • Search Google Scholar
    • Export Citation
  • 14.

    Pegram R, Clifford C, Walker J, Keirans J, 1987. Clarification of the Rhipicephalus sanguineus group (Acari, Ixodoidea, Ixodidae). I. R. sulcatus Neumann, 1908 and R. turanicus Pomerantsev, 1936. Syst Parasitol 10: 326.

    • Search Google Scholar
    • Export Citation
  • 15.

    Walker J, Keirans J, Horak I, 2000. The genus Rhipicephalus (Acari, Ixoidae). A Guide to the Brown Ticks of the World. Cambridge: Cambridge University Press.

    • Search Google Scholar
    • Export Citation
  • 16.

    Feldman-Muhsam B, 1954. Revision of the genus Hyalomma. Bull Res Counc Isr 64: 150170.

  • 17.

    Leitner M, Yitzhaki S, Rzotkiewicz S, Keysary A, 2002. Polymerase chain reaction-based diagnosis of Mediterranean spotted fever in serum and tissue samples. Am J Trop Med Hyg 67: 166169.

    • Search Google Scholar
    • Export Citation
  • 18.

    Eremeeva ME, Bosserman EA, Demma LJ, Zambrano ML, Blau DM, Dasch GA, 2006. Isolation and identification of Rickettsia massiliae from Rhipicephalus sanguineus ticks collected in Arizona. Appl Environ Microbiol 72: 55695577.

    • Search Google Scholar
    • Export Citation
  • 19.

    Sekeyova Z, Roux V, Raoult D, 2001. Phylogeny of Rickettsia spp. inferred by comparing sequences of ‘gene D', which encodes an intracytoplasmic protein. Int J Syst Evol Microbiol 51: 13531360.

    • Search Google Scholar
    • Export Citation
  • 20.

    Roux V, Raoult D, 2000. Phylogenetic analysis of members of the genus Rickettsia using the gene encoding the outer-membrane protein rOmpB (ompB). Int J Syst Evol Microbiol 50: 14491455.

    • Search Google Scholar
    • Export Citation
  • 21.

    Tamura K, Dudley J, Nei M, Kumar S, 2007. MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol 24: 15961599.

    • Search Google Scholar
    • Export Citation
  • 22.

    Eremeeva ME, Stromdahl EY, 2011. Short report: new spotted fever group Rickettsia in a Rhipicephalus turanicus tick removed from a child in eastern Sicily, Italy. Am J Trop Med Hyg 84: 99101.

    • Search Google Scholar
    • Export Citation
  • 23.

    Sarih M, Socolovschi C, Boudebouch N, Hassar M, Raoult D, Parola P, 2008. Spotted fever group rickettsiae in ticks, Morocco. Emerg Infect Dis 14: 10671073.

    • Search Google Scholar
    • Export Citation
  • 24.

    Raoult D, Fournier PE, Abboud P, Caron F, 2002. First documented human Rickettsia aeschlimannii infection. Emerg Infect Dis 8: 748749.

  • 25.

    Parola P, 2006. Rickettsioses in sub-Saharan Africa. Ann N Y Acad Sci 1078: 4247.

  • 26.

    Shpynov S, Fournier PE, Rudakov N, Tankibaev M, Tarasevich I, Raoult D, 2004. Detection of a Rickettsia closely related to Rickettsia aeschlimannii, “Rickettsia heilongjiangensis,” Rickettsia sp. strain RpA4, and Ehrlichia muris in ticks collected in Russia and Kazakhstan. J Clin Microbiol 42: 22212223.

    • Search Google Scholar
    • Export Citation
  • 27.

    Portillo A, Santibanez P, Santibanez S, Perez-Martinez L, Oteo JA, 2008. Detection of Rickettsia spp. in Haemaphysalis ticks collected in La Rioja, Spain. Vector Borne Zoonotic Dis 8: 653658.

    • Search Google Scholar
    • Export Citation
  • 28.

    Mura A, Socolovschi C, Ginesta J, Lafrance B, Magnan S, Rolain JM, Davoust B, Raoult D, Parola P, 2008. Molecular detection of spotted fever group rickettsiae in ticks from Ethiopia and Chad. Trans R Soc Trop Med Hyg 102: 945949.

    • Search Google Scholar
    • Export Citation
  • 29.

    Fernandez-Soto P, Diaz Martin V, Perez-Sanchez R, Encinas-Grandes A, 2009. Increased prevalence of Rickettsia aeschlimannii in Castilla y Leon, Spain. Eur J Clin Microbiol Infect Dis 28: 693695.

    • Search Google Scholar
    • Export Citation
  • 30.

    Bitam I, Parola P, Matsumoto K, Rolain JM, Baziz B, Boubidi SC, Harrat Z, Belkaid M, Raoult D, 2006. First molecular detection of R. conorii, R. aeschlimannii, and R. massiliae in ticks from Algeria. Ann N Y Acad Sci 1078: 368372.

    • Search Google Scholar
    • Export Citation
  • 31.

    Loftis AD, Reeves WK, Szumlas DE, Abbassy MM, Helmy IM, Moriarity JR, Dasch GA, 2006. Rickettsial agents in Egyptian ticks collected from domestic animals. Exp Appl Acarol 40: 6781.

    • Search Google Scholar
    • Export Citation
  • 32.

    Psaroulaki A, Ragiadakou D, Kouris G, Papadopoulos B, Chaniotis B, Tselentis Y, 2006. Ticks, tick-borne rickettsiae, and Coxiella burnetii in the Greek Island of Cephalonia. Ann N Y Acad Sci 1078: 389399.

    • Search Google Scholar
    • Export Citation
  • 33.

    Fernandez-Soto P, Encinas-Grandes A, Perez-Sanchez R, 2003. Rickettsia aeschlimannii in Spain: molecular evidence in Hyalomma marginatum and five other tick species that feed on humans. Emerg Infect Dis 9: 889890.

    • Search Google Scholar
    • Export Citation
  • 34.

    Pretorius AM, Birtles RJ, 2002. Rickettsia aeschlimannii: a new pathogenic spotted fever group Rickettsia, South Africa. Emerg Infect Dis 8: 874.

    • Search Google Scholar
    • Export Citation
  • 35.

    Brouqui P, Parola P, Fournier PE, Raoult D, 2007. Spotted fever rickettsioses in southern and eastern Europe. FEMS Immunol Med Microbiol 49: 212.

    • Search Google Scholar
    • Export Citation
  • 36.

    Letaief A, 2006. Epidemiology of rickettsioses in North Africa. Ann N Y Acad Sci 1078: 3441.

  • 37.

    Parola P, Socolovschi C, Jeanjean L, Bitam I, Fournier PE, Sotto A, Labauge P, Raoult D, 2008. Warmer weather linked to tick attack and emergence of severe rickettsioses. PLoS Negl Trop Dis 2: e338.

    • Search Google Scholar
    • Export Citation
  • 38.

    Garcia-Garcia JC, Portillo A, Nunez MJ, Santibanez S, Castro B, Oteo JA, 2010. A patient from Argentina infected with Rickettsia massiliae. Am J Trop Med Hyg 82: 691692.

    • Search Google Scholar
    • Export Citation
  • 39.

    Vitale G, Mansuelo S, Rolain JM, Raoult D, 2006. Rickettsia massiliae human isolation. Emerg Infect Dis 12: 174175.

  • 40.

    Rolain JM, Maurin M, Vestris G, Raoult D, 1998. In vitro susceptibilities of 27 rickettsiae to 13 antimicrobials. Antimicrob Agents Chemother 42: 15371541.

    • Search Google Scholar
    • Export Citation
  • 41.

    Raoult D, Fournier PE, Eremeeva M, Graves S, Kelly PJ, Oteo JA, Sekeyova Z, Tamura A, Tarasevich I, Zhang L, 2005. Naming of Rickettsiae and rickettsial diseases. Ann N Y Acad Sci 1063: 112.

    • Search Google Scholar
    • Export Citation
  • 42.

    Murray RG, Stackebrandt E, 1995. Taxonomic note: implementation of the provisional status Candidatus for incompletely described procaryotes. Int J Syst Bacteriol 45: 186187.

    • Search Google Scholar
    • Export Citation
  • 43.

    Goldwasser RA, Steiman Y, Klingberg W, Swartz TA, Klingberg MA, 1974. The isolation of strains of rickettsiae of the spotted fever group in Israel and their differentiation from other members of the group by immunofluorescence methods. Scand J Infect Dis 6: 5362.

    • Search Google Scholar
    • Export Citation

Author Notes

*Address correspondence to Avi Keysary, Department of Infectious Diseases, Israel Institute for Biological Research, P.O. Box 19, 74100, Ness Ziona, Israel. E-mail: rickiticki6@gmail.com

Authors' addresses: Avi Keysary, Department of Infectious Diseases, Israel Institute for Biological Research, Ness Ziona, Israel, E-mail: rickiticki6@gmail.com. Marina E. Eremeeva, Rickettsial Zoonones Branch, Centers for Disease Control and Prevention, Atlanta, GA, E-mail: m.eremeeva01@gmail.com. Moshe Leitner and Adi Beth Din, Department of Biochemistry, Israel Institute for Biological Research, Ness Ziona, Israel, E-mails: moshel@iibr.gov.il and adib@iibr.gov.il. Mary E. Wikswo, Division of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, GA, E-mail: ezq1@cdc.gov. Kosta Y. Mumcuoglu, Department of Microbiology and Molecular Genetics, Hebrew University-Hadassah Medical School, Jerusalem, Israel, E-mail: kostam@cc.huji.ac.il. Moshe Inbar, Arian D. Wallach, and Uri Shanas, Department of Evolutionary and Environmental Biology, University of Haifa, Mount Carmel, Haifa, Israel, E-mails: minbar@research.haifa.ac.il, arian.wallach@bigpond.com, and shanas@research.haifa.ac.il. Roni King, Israel Nature and Parks Authority, Jerusalem, E-mail: king@npa.org.il. Trevor Waner, Animal Facilities, Israel Institute for Biological Research, Ness Ziona, Israel, E-mail: wanertnt@gmail.com

Save