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Abstract
Q-fever is a zoonosis caused by the gram-negative obligate intracellular pathogen Coxiella burnetii. Since its discovery, and particularly following the recent outbreaks in the Netherlands, C. burnetii appeared as a clear public health concern. In the present study, the infectious potential displayed by goat and cattle isolates of C. burnetii was compared to a reference strain (Nine Mile) using both in vitro (human HeLa and bovine macrophage cells) and in vivo (BALB/c mice) models. The isolates had distant genomic profiles with one - the goat isolate - being identical to the predominant strain circulating in the Netherlands during the 2007–2010 outbreaks. Infective doses were established with ethidium monoazide-PCR for the first time here applied to C. burnetii. This method allowed for the preparation of reproducible and characterized inocula thanks to its capacity to discriminate between live and dead cells. Globally, the proliferative capacity of the Nine Mile strain in cell lines and mice was higher compared to the newly isolated field strains. In vitro, the bovine C. burnetii isolate multiplied faster in a bovine macrophage cell line, an observation tentatively explained by the preferential specificity of this strain for allogeneic host cells. In the BALB/c mouse model, however, the goat and bovine isolates multiplied at about the same rate indicating no peculiar hypervirulent behavior in this animal model.
Highlights
Q-fever is a zoonosis caused by Coxiella burnetii, an intracellular gram-negative bacterium
Cell Lines and Culture Media Human carcinoma HeLa cells were cultivated in Dulbecco modified Eagle medium (DMEM) supplemented with 10% foetal calf serum (FCS) and 0.1 mM of nonessential amino acids solution
The complete genotype of these strains was obtained by both MLVA and a single nucleotide polymorphism typing scheme [9,30,31] and were compared to that of the reference strain Nine Mile phase I (NMI)
Summary
Q-fever is a zoonosis caused by Coxiella burnetii, an intracellular gram-negative bacterium. Q-fever manifests in humans as acute or chronic illness. Acute Q-fever ranges from an asymptomatic state to an abrupt, flu-like illness that can be accompanied with high fever, general malaise, pneumonia, myalgia and hepatitis. Chronic Q-fever occurs in individuals in approximately 1–5% of cases, and typically manifests as endocarditis [1,2]. Humans contract the disease mainly by inhalation of airborne particles contaminated with C. burnetii (feces, milk or birth products -placenta and amniotic fluid-) of infected animals [3,4]. Up to 109 C. burnetii cells per gram of placenta can be excreted [5]. Considering the infective dose of this bacterium has been reported to be close to one [6], these products are obviously hazardous to humans
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