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. 2021 Feb 26:10:625576.
doi: 10.3389/fcimb.2020.625576. eCollection 2020.

Phylogeography of Human and Animal Coxiella burnetii Strains: Genetic Fingerprinting of Q Fever in Belgium

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Phylogeography of Human and Animal Coxiella burnetii Strains: Genetic Fingerprinting of Q Fever in Belgium

Sara Tomaiuolo et al. Front Cell Infect Microbiol. .

Abstract

Q fever is a zoonotic disease caused by the bacteria Coxiella burnetii. Domestic ruminants are the primary source for human infection, and the identification of likely contamination routes from the reservoir animals the critical point to implement control programs. This study shows that Q fever is detected in Belgium in abortion of cattle, goat and sheep at a different degree of apparent prevalence (1.93%, 9.19%, and 5.50%, respectively). In addition, and for the first time, it is detected in abortion of alpaca (Vicugna pacos), raising questions on the role of these animals as reservoirs. To determine the relationship between animal and human strains, Multiple Locus Variable-number Tandem Repeat Analysis (MLVA) (n=146), Single-Nucleotide Polymorphism (SNP) (n=92) and Whole Genome Sequencing (WGS) (n=4) methods were used to characterize samples/strains during 2009-2019. Three MLVA clusters (A, B, C) subdivided in 23 subclusters (A1-A12, B1-B8, C1-C3) and 3 SNP types (SNP1, SNP2, SNP6) were identified. The SNP2 type/MLVA cluster A was the most abundant and dispersed genotype over the entire territory, but it seemed not responsible for human cases, as it was only present in animal samples. The SNP1/MLVA B and SNP6/MLVA C clusters were mostly found in small ruminant and human samples, with the rare possibility of spillovers in cattle. SNP1/MLVA B cluster was present in all Belgian areas, while the SNP6/MLVA C cluster appeared more concentrated in the Western provinces. A broad analysis of European MLVA profiles confirmed the host-species distribution described for Belgian samples. In silico genotyping (WGS) further identified the spacer types and the genomic groups of C. burnetii Belgian strains: cattle and goat SNP2/MLVA A isolates belonged to ST61 and genomic group III, while the goat SNP1/MLVA B strain was classified as ST33 and genomic group II. In conclusion, Q fever is widespread in all Belgian domestic ruminants and in alpaca. We determined that the public health risk in Belgium is likely linked to specific genomic groups (SNP1/MLVA B and SNP6/MLVA C) mostly found in small ruminant strains. Considering the concordance between Belgian and European results, these considerations could be extended to other European countries.

Keywords: Coxiella burnetii; MLVA; SNP; WGS; alpaca; animals; humans.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Minimum spanning trees showing the relationship between Multiple Locus Variable-number Tandem Repeat Analysis (MLVA) genotypes of Belgian isolates collected between 2010 and 2019. (A) Animal and human positive clinical samples with a complete 13-locus MLVA profile or missing max two out of 13 markers (n=114) were included in the clustering together with the reference strain profiles (n=8) obtained from published data (Roest et al., 2011; Joulié et al., 2017). Each circle corresponds to a single MLVA profile and the size of the circle is proportional to the number of samples sharing an identical genotype. The gray background connects samples differing of only one marker from each other. Homogeneous bovine and caprine genotypes are grouped in the core of the tree, while heterogeneous caprine and human profiles occupy the two lateral branches. (B) Minimum spanning tree resulting from the collapsing of (A) in which nodes differing of only one marker were merged together, as they may represent microvariants of the same genotype. This analysis separated 13-locus MLVA profiles in three main clusters (A, B, C) and different sub-clusters (A1-A12, B1-B8, C1-C3).
Figure 2
Figure 2
Comparison between (A) Single-Nucleotide Polymorphism (SNP) and (B) Multiple Locus Variable-number Tandem Repeat Analysis (MLVA) clustering. (A) Ninety-three Belgian clinical samples positive for C. burnetii were clustered basing on their SNP profile. (B) Fifty-two samples typed by SNP and MLVA were clustered by minimum spanning tree built on 13-locus MLVA profiles (complete or lacking max. two markers). MLVA profiles corresponding to a unique SNP genomic group are surrounded by a dotted line. Each circle corresponds to a single MLVA profile and the size of the circle is proportional to the number of samples sharing an identical genotype. The gray background connects samples differing of only one marker from each other.
Figure 3
Figure 3
Geographical distribution of Belgian C. burnetii strains based on (A) Multiple Locus Variable-number Tandem Repeat Analysis (MLVA) and (B) Single-Nucleotide Polymorphism (SNP) genotypes detected between 2010 and 2019. (A) Localization of the MLVA profiles according to the clusters identified in Figure 1B and to the host species. (B) Distribution of C. burnetii positive samples according to the SNP genotypes ( Table S2 ) and the host species.
Figure 4
Figure 4
Minimum spanning trees showing the clustering of 855 Multiple Locus Variable-number Tandem Repeat Analysis (MLVA) profiles from European positive samples according to (A) the host species and (B) the country of origin. Data were collected from the present study, the MLVA online database and published studies. The microsatellite 7-locus MLVA panel was used for the clustering including only complete profiles, or missing max. two out of seven markers. Each circle corresponds to a single MLVA profile and the size of the circle is proportional to the number of samples sharing an identical genotype. The gray background connects samples differing of only one marker from each other.

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References

    1. Agerholm J. S. (2013). Coxiella burnetii associated reproductive disorders in domestic animals-a critical review. Acta Vet. Scand. 55, 13. 10.1186/1751-0147-55-13 - DOI - PMC - PubMed
    1. Arricau Bouvery N., Souriau A., Lechopier P., Rodolakis A. (2003). Experimental Coxiella burnetii infection in pregnant goats: excretion routes. Vet. Res. 34, 423–433. 10.1051/vetres:2003017 - DOI - PubMed
    1. Arricau-Bouvery N., Rodolakis A. (2005). Is Q fever an emerging or re-emerging zoonosis? Vet. Res. 36, 327–349. 10.1051/vetres:2005010 - DOI - PubMed
    1. Arricau-Bouvery N., Hauck Y., Bejaoui A., Frangoulidis D., Bodier C. C., Souriau A., et al. . (2006). Molecular characterization of Coxiella burnetii isolates by infrequent restriction site-PCR and MLVA typing. BMC Microbiol. 6, 38. 10.1186/1471-2180-6-38 - DOI - PMC - PubMed
    1. Astobiza I., Tilburg J. J. H. C., Piñero A., Hurtado A., García-Pérez A. L., Nabuurs-Franssen M. H., et al. . (2012). Genotyping of Coxiella burnetii from domestic ruminants in northern Spain. BMC Vet. Res. 8, 241. 10.1186/1746-6148-8-241 - DOI - PMC - PubMed

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