Three-Toed Sloth as Putative Reservoir of Coxiella burnetii, Cayenne, French Guiana

Bernard Davoust (1) , Jean-Lou Marié (2, 1) , Vincent Pommier de Santi (3, 4) , Jean Michel Berenger (5, 1) , Sophie Edouard (1) , Didier Raoult (5, 1)

1 AMU - Aix Marseille Université
2 Service de Santé des Armées
3 Centre d'épidémiologie et de santé publique des armées
4 Direction Interarmées du Service de Santé en Guyane
5 URMITE - Unité de Recherche sur les Maladies Infectieuses et Tropicales Emergentes

To the Editor: Q fever is an emerging zoonosis and a major public health concern in French Guiana, a French overseas region located on the northeastern coast of South America (1,2). Most cases occur in the city of Cayenne (3), specifically in the suburbs , where houses are near wooded hills (4). Genotyping performed by using multispacer sequence typing showed that MST17, a unique genotype of C. burnetii, circulates in Cay-enne and is responsible for epidemics of Q fever (5). C. burnetii transmission peaks during the rainy season, and the incidence of Q fever usually increases 1–3 months later (6). The animal reservoir of C. burnetii in French Guiana is unknown; previous studies have excluded domestic ruminants, which are known to be C. burnetii reservoirs elsewhere in the world (6). Four sero-logic surveys showed few C. burnetii– positive opossums, dogs, rodents (Proechimys spp.), bovines, or birds in French Guiana (7). In 2013, using real-time PCR (qPCR) analysis of vaginal swab samples, we showed that 6/158 (3.8%) dogs from Cayenne and 0/206 bats from the coastal area of French Guiana were positive for C. burnetii (Cycle threshhold [C t ]<35). One of the positive samples was identified as genotype MST17 (5). A case–control study among humans identified several risk factors for Q fever, including living near a forest and the presence of wild animals near the house (6). During January–April 2013, a Q fever outbreak occurred in Tiger Camp, a military residential area located at the top of a wooded hill in Cay-enne. Vaginal swab samples were collected from animals living in the area (13 goats, 8 sheep, 7 bats, 34 birds, 2 opossums, 4 iguanas, and 17 geckos); all samples were negative for C. bur-netii by qPCR. In addition, serologic tests for C. burnetii were negative for samples from all 37 small ruminants maintained near the outbreak area. In January 2014, a dead (accidental death) female 3-toed sloth (Brad-ypus tridactylus) (Figure, panel A) was found on the road near the residence of a Q fever patient. We retrieved the sloth and collected feces, spleen, liver, kidney, lung, and uterus samples and a vaginal swab sample. A total of 16 ticks were removed from the sloth and stored in 70% alcohol. DNA was extracted from the fe-ces, organs, and ticks by using the BioRobot EZ1 Workstation (QIA-GEN, Courtaboeuf, France). qPCR targeting the repeated insertion sequence IS1111 was performed by using a CFX96 Touch Real-Time PCR Detection System (Bio-Rad, Marne la Coquette, France) as described (8). We confirmed all positive results by performing a second qPCR targeting the IS30a repeated sequence. DNA samples with C t values <35 in both assays were considered positive for C. burnetii. A standard calibration curve quantifying the target IS1111 was generated by using 10-fold serial dilutions of C. burnetii Nine Mile strain. The number of IS1111 intergenic sequences found in the genome of strain C. burnetii MST17 was identical to that for the Nine Mile strain (F. D'Amato, unpub. data); thus, the qPCR that we used was valid for quantifying the number of C. burnetii MST17 IS1111 copies/mL in samples we collected (5). qPCR analysis showed that the feces were highly positive for C. bur-netii; the sample had a low C t value of 23, corresponding to 7 log 10 DNA copies/mL (9). The spleen was also positive for C. burnetii; the C t value was 34, corresponding to 3.6 log 10 DNA copies/mL. Results for the other samples were negative. Using morphologic criteria, we identified all 16 ticks collected from the sloth as Amblyomma geayi (Figure,LETTERSpanelB).WeperformedC.burnetii–specificqPCRontheticks;14(88%)werepositive.WegenotypedC.burnetii–positiveDNAfromthefecesandfrom6ofthe16ticksbyusingmultispacersequencetypingasdescribed(5).AllsampleswereidentifiedasMST17,theuniquegenotypecirculatinginCayenne(5).Afterobtainingthelaboratoryre-sults,weconfirmedthatalocalgroupinchargeofthecollectionandtreatmentofinjuredanimalsusuallyreleasedrehabilitated3-toedslothsintoTigerCamp.ResidentsofTigerCampregu-larlyobservedandcameintocontactwiththesloths,andtickswerefrequent-lyobservedonthefuroftheanimals.Furthermore,3QfeverpatientsfromCayennereportedcontactwithsloths.FecesfromtheslothinthisstudywerehighlyinfectiousforC.burnetii.Becauseslothsliveintalltreesandcanshedthisbacteriumintheirfeces,humancontaminationmightoccurthroughin-halationofinfectiousaerosolsfromfe-ces.ThehighprevalenceofC.burnetiiinfectioninticksalsosuggestspossibletransmissionthroughtickbitesorfromaerosolsoftickfecesthathavebeende-positedontheskinofanimalhosts;suchfecescanbeextremelyrichinbacteriaandhighlyinfectious(10).Inthis2013outbreakofQfever,epidemiologicstudiesledtotheiden-tificationof3-toedslothsasaputativesourceofC.burnetiiinfection.FurtherinvestigationsareneededtoconfirmtheroleofslothsasareservoirforC.burnetiiinFrenchGuianaandtoim-plementefficientmeasurestopreventtransmissiontohumans.

Mots clés: Sloth; Putative Reservoir; Coxiella burnetii; Cayenne

Domaines: Médecine humaine et pathologie

Bernard Davoust, Jean-Lou Marié, Vincent Pommier de Santi, Jean Michel Berenger, Sophie Edouard, et al.. Three-Toed Sloth as Putative Reservoir of Coxiella burnetii, Cayenne, French Guiana . Emerging Infectious Diseases, 2014, 20, pp.1760-1761. ⟨hal-01245508⟩