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

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.

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Molecular detection of Coxiella burnetii in small ruminants and genotyping of specimens collected from goats in Poland

BMC Veterinary Research ◽

10.1186/s12917-021-03051-0 ◽

2021 ◽

Vol 17 (1) ◽

Author(s):

Agnieszka Jodełko ◽

Monika Szymańska-Czerwińska ◽

Jolanta Grażyna Rola ◽

Krzysztof Niemczuk

Keyword(s):

Coxiella Burnetii ◽

Small Ruminant ◽

Q Fever ◽

Variable Number Tandem Repeat ◽

Small Ruminants ◽

Variable Number ◽

Tissue Sections ◽

Pcr Targeting ◽

Sequence Types ◽

Multispacer Sequence Typing

Abstract Background Coxiella burnetii is the etiological agent of Q fever, a zoonosis affecting many animal species including sheep and goats. The aims of this study were to evaluate the shedding of Coxiella burnetii in small ruminant herds and to identify the pathogen’s genotypes and sequence types (STs) using multiple-locus variable number tandem repeat analysis (MLVA) and multispacer sequence typing (MST) methods. Results Overall, 165 samples from 43 herds of goats and 9 flocks of sheep were collected including bulk tank milk (BTM), individual milk samples, vaginal swabs, tissue sections from stillborn kids, feces and placentas. These were tested by real-time PCR targeting the IS1111 element. C. burnetii infection was confirmed in 51.16% of the herds of goats and 22.2% of the flocks of sheep. Six out of nine samples originating from goats were successfully genotyped using the MLVA method. The presence was confirmed of two widely distributed MLVA genotypes (I and J) and genotype PL1 previously reported only in cattle. Only one sequence type (ST61) was identified; however, the majority of specimens represented partial STs and some of them may belong to ST61. Other partial STs could possibly be ST74. Conclusion This study confirmed the relatively common occurrence of Coxiella burnetii in small ruminant herds in Poland. Interestingly, all genotyped samples represent cattle-associated MLVA genotypes.

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Coxiella burnetii Shedding in Milk and Molecular Typing of Strains Infecting Dairy Cows in Greece

Pathogens ◽

10.3390/pathogens10030287 ◽

2021 ◽

Vol 10 (3) ◽

pp. 287

Author(s):

Emmanouil Kalaitzakis ◽

Tiziano Fancello ◽

Xavier Simons ◽

Ilias Chaligiannis ◽

Sara Tomaiuolo ◽

...

Keyword(s):

Dairy Cows ◽

Coxiella Burnetii ◽

Q Fever ◽

Variable Number Tandem Repeat ◽

Variable Number ◽

Human Infection ◽

Animal Reservoir

Ruminants are considered the commonest animal reservoir for human infection of Coxiella burnetii, the Q fever causative agent. Considering the recently described importance of human Q fever in Greece, we aimed at providing the first comprehensive direct evidence of C. burnetii in dairy cows in Greece, including the genetic characterization of strains. The 462 examined dairy farms represented all geographical areas of Greece. One bulk tank milk sample was collected from every farm and tested for the presence of C. burnetii. Molecular genotyping of strains, performed directly on samples, revealed the existence of two separate clades characterized by single nucleotide polymorphism (SNP) genotypes of type 1 and type 2. The two clades were clearly distinguished in multiple locus variable-number tandem repeat analysis (MLVA) by two discriminative loci: MS30 and MS28. Whereas MLVA profiles of SNP-type 2 clade were closely related to strains described in other European cattle populations, the MLVA profile observed within the SNP type 1 clade highlighted a peculiar genetic signature for Greece, related to genotypes found in sheep and goats in Europe. The shedding of C. burnetii bearing this genotype might have yet undefined human epidemiological consequences. Surveillance of the genetic distribution of C. burnetii from different sources is needed to fully understand the epidemiology of Q fever in Greece.

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First Complete Genome Sequence of the Dutch VeterinaryCoxiella burnetiiStrain NL3262, Originating from the Largest Global Q Fever Outbreak, and Draft Genome Sequence of Its Epidemiologically Linked Chronic Human Isolate NLhu3345937

Genome Announcements ◽

10.1128/genomea.00245-16 ◽

2016 ◽

Vol 4 (2) ◽

Cited By ~ 9

Author(s):

Runa Kuley ◽

Hilde E. Smith ◽

Ingmar Janse ◽

Frank L. Harders ◽

Frank Baas ◽

...

Keyword(s):

Genome Sequence ◽

Coxiella Burnetii ◽

Complete Genome Sequence ◽

Complete Genome ◽

Q Fever ◽

Draft Genome ◽

Variable Number Tandem Repeat ◽

Variable Number ◽

Outbreak Strain ◽

Repeat Analysis

The largest global Q fever outbreak occurred in The Netherlands during 2007 to 2010. Goats and sheep were identified as the major sources of disease. Here, we report the first complete genome sequence ofCoxiella burnetiigoat outbreak strain NL3262 and that of an epidemiologically linked chronic human strain, both having the outbreak-relatedCbNL01multilocus variable-number tandem-repeat analysis (MLVA) genotype.

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Correlating Genotyping Data of Coxiella burnetii with Genomic Groups

Pathogens ◽

10.3390/pathogens10050604 ◽

2021 ◽

Vol 10 (5) ◽

pp. 604

Author(s):

Claudia M. Hemsley ◽

Angela Essex-Lopresti ◽

Isobel H. Norville ◽

Richard W. Titball

Keyword(s):

Coxiella Burnetii ◽

Classification Scheme ◽

Q Fever ◽

Variable Number Tandem Repeat ◽

Febrile Illness ◽

Variable Number ◽

New Classification ◽

Genetic Lineages ◽

Virulence Potential ◽

Multispacer Sequence Typing

Coxiella burnetii is a zoonotic pathogen that resides in wild and domesticated animals across the globe and causes a febrile illness, Q fever, in humans. Several distinct genetic lineages or genomic groups have been shown to exist, with evidence for different virulence potential of these lineages. Multispacer Sequence Typing (MST) and Multiple-Locus Variable number tandem repeat Analysis (MLVA) are being used to genotype strains. However, it is unclear how these typing schemes correlate with each other or with the classification into different genomic groups. Here, we created extensive databases for published MLVA and MST genotypes of C. burnetii and analysed the associated metadata, revealing associations between animal host and human disease type. We established a new classification scheme that assigns both MST and MLVA genotypes to a genomic group and which revealed additional sub-lineages in two genomic groups. Finally, we report a novel, rapid genomotyping method for assigning an isolate into a genomic group based on the Cox51 spacer sequence. We conclude that by pooling and streamlining existing datasets, associations between genotype and clinical outcome or host source were identified, which in combination with our novel genomotyping method, should enable an estimation of the disease potential of new C. burnetii isolates.

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Circulation of Coxiella burnetii in a Naturally Infected Flock of Dairy Sheep: Shedding Dynamics, Environmental Contamination, and Genotype Diversity

Applied and Environmental Microbiology ◽

10.1128/aem.02180-15 ◽

2015 ◽

Vol 81 (20) ◽

pp. 7253-7260 ◽

Cited By ~ 28

Author(s):

A. Joulié ◽

K. Laroucau ◽

X. Bailly ◽

M. Prigent ◽

P. Gasqui ◽

...

Keyword(s):

Coxiella Burnetii ◽

Environmental Contamination ◽

Q Fever ◽

Management Strategies ◽

Variable Number Tandem Repeat ◽

Variable Number ◽

Dairy Sheep ◽

Content Type ◽

Ewe Lambs ◽

Genotype Diversity

ABSTRACTQ fever is a worldwide zoonosis caused byCoxiella burnetii. Domestic ruminants are considered to be the main reservoir. Sheep, in particular, may frequently cause outbreaks in humans. Because within-flock circulation data are essential to implementing optimal management strategies, we performed a follow-up study of a naturally infected flock of dairy sheep. We aimed to (i) describeC. burnetiishedding dynamics by sampling vaginal mucus, feces, and milk, (ii) assess circulating strain diversity, and (iii) quantify barn environmental contamination. For 8 months, we sampled vaginal mucus and feces every 3 weeks from aborting and nonaborting ewes (n= 11 andn= 26, respectively); for lactating females, milk was obtained as well. We also sampled vaginal mucus from nine ewe lambs. Dust and air samples were collected every 3 and 6 weeks, respectively. All samples were screened using real-time PCR, and strongly positive samples were further analyzed using quantitative PCR. Vaginal and fecal samples with sufficient bacterial burdens were then genotyped by multiple-locus variable-number tandem-repeat analysis (MLVA) using 17 markers.C. burnetiiburdens were higher in vaginal mucus and feces than in milk, and they peaked in the first 3 weeks postabortion or postpartum. Primiparous females and aborting females tended to shedC. burnetiilonger and have higher bacterial burdens than nonaborting and multiparous females. Six genotype clusters were identified; they were independent of abortion status, and within-individual genotype diversity was observed.C. burnetiiwas also detected in air and dust samples. Further studies should determine whether the within-flock circulation dynamics observed here are generalizable.

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Evaluation of the Diagnostic Potential of Recombinant Coxiella burnetii Com1 in an ELISA for the Diagnosis of Q Fever in Sheep, Goats and Cattle

Microorganisms ◽

10.3390/microorganisms8081235 ◽

2020 ◽

Vol 8 (8) ◽

pp. 1235 ◽

Cited By ~ 1

Author(s):

Mareike Stellfeld ◽

Claudia Gerlach ◽

Ina-Gabriele Richter ◽

Peter Miethe ◽

Dominika Fahlbusch ◽

...

Keyword(s):

Sensitivity And Specificity ◽

Coxiella Burnetii ◽

Q Fever ◽

Roc Curves ◽

Diagnostic Tools ◽

Enzyme Linked Immunosorbent Assay ◽

Domestic Ruminants ◽

Diagnostic Potential ◽

Assay Performance ◽

Higher Sensitivity

Coxiella burnetii is the causative agent of Q fever, a zoonosis infecting domestic ruminants and humans. Currently used routine diagnostic tools offer limited sensitivity and specificity and symptomless infected animals may be missed. Therefore, diagnostic tools of higher sensitivity and specificity must be developed. For this purpose, the C. burnetii outer membrane protein Com1 was cloned and expressed in Escherichia coli. The His-tagged recombinant protein was purified and used in an indirect enzyme-linked immunosorbent assay (ELISA). Assay performance was tested with more than 400 positive and negative sera from sheep, goats and cattle from 36 locations. Calculation of sensitivity and specificity was undertaken using receiver operating characteristic (ROC) curves. The sensitivities and specificities for sheep were 85% and 68% (optical density at 450nm, OD450 cut-off value 0.32), for goats 94% and 77% (OD450 cut-off value 0.23) and for cattle 71% and 70% (OD450 cut-off value 0.18), respectively. These results correspond to excellent, outstanding and acceptable discrimination of positive and negative sera. In summary, recombinant Com1 can provide a basis for more sensitive and specific diagnostic tools in veterinary medicine.

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The prevalence of Coxiella burnetii in ticks and animals in Slovenia

BMC Veterinary Research ◽

10.1186/s12917-019-2130-3 ◽

2019 ◽

Vol 15 (1) ◽

Cited By ~ 4

Author(s):

Nataša Knap ◽

Diana Žele ◽

Urška Glinšek Biškup ◽

Tatjana Avšič-Županc ◽

Gorazd Vengušt

Keyword(s):

Coxiella Burnetii ◽

Q Fever ◽

Primary Source ◽

Domestic Animals ◽

Animal Blood ◽

Blood Samples ◽

Specific Antibodies ◽

Source Of Infection ◽

History Of ◽

Pathogen Dna

Abstract Background The obligate intracellular bacterium Coxiella burnetii causes globally distributed zoonotic Q fever. Ruminant livestock are common reservoirs of C. burnetii. Coxiella burnetii are shed in large numbers in the waste of infected animals and are transmitted by inhalation of contaminated aerosols. This study was conducted to evaluate the prevalence of C. burnetii infection in domestic animals and ticks in areas of Slovenia associated with a history of Q fever outbreaks. Results A total of 701 ticks were collected and identified from vegetation, domestic animals and wild animals. C. burnetii DNA was detected in 17 out of 701 (2.4%) ticks. No C. burnetii DNA was found in male ticks. Ticks that tested positive in the PCR-based assay were most commonly sampled from wild deer (5.09%), followed by ticks collected from domestic animals (1.16%) and ticks collected by flagging vegetation (0.79%). Additionally, 150 animal blood samples were investigated for the presence of C. burnetii-specific antibodies and pathogen DNA. The presence of pathogen DNA was confirmed in 14 out of 150 (9.3%) blood samples, while specific antibodies were detected in sera from 60 out of 150 (40.4%) animals. Conclusions Our results indicate that ticks, although not the primary source of the bacteria, are infected with C. burnetii and may represent a potential source of infection for humans and animals. Ticks collected from animals were most likely found to harbor C. burnetii DNA, and the infection was not lost during molting. The persistence and distribution of pathogens in cattle and sheep indicates that C. burnetii is constantly present in Slovenia.

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Q fever in Quebec (1989–93): Report of 14 Cases

Canadian Journal of Infectious Diseases ◽

10.1155/1994/531807 ◽

1994 ◽

Vol 5 (3) ◽

pp. 113-118 ◽

Cited By ~ 2

Author(s):

Monique Goyette ◽

André Poirier ◽

Jean Bouchard ◽

Eric Morrier

Keyword(s):

Coxiella Burnetii ◽

Q Fever ◽

Serological Screening ◽

Rural Region ◽

Domestic Ruminants ◽

Animal Populations ◽

The World

Q fever, a zoonosis acquired by inhalation of the rickettsiaCoxiella burnetii, is rarely diagnosed in Canada. The world incidence has been increasing since 1960, because of progressive dissemination of this microorganism in animal populations, particularly domestic ruminants. Some recent outbreaks were caused by cats. Of 14 cases reported in Quebec between 1989 and the beginning of 1993, nine occurred successively in an 18-month period in the rural region surrounding Trois-Rivières, after contact with livestock or cats. These cases are reported here, with the results of serological screening of the workers of an abattoir where one of the cases worked. Five additional cases reported in Quebec during the same period are briefly reviewed.

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Molecular detection of Coxiella burnetii infection in aborted samples of domestic ruminants in Iran

PLoS ONE ◽

10.1371/journal.pone.0250116 ◽

2021 ◽

Vol 16 (4) ◽

pp. e0250116

Author(s):

Ashraf Mohabati Mobarez ◽

Mohammad Khalili ◽

Ehsan Mostafavi ◽

Saber Esmaeili

Keyword(s):

Coxiella Burnetii ◽

Real Time Pcr ◽

Molecular Detection ◽

Q Fever ◽

Domestic Animal ◽

Domestic Ruminant ◽

Domestic Ruminants ◽

Pcr Targeting ◽

Different Parts

Background Coxiella burnetii is the causative agent of Q fever which is a highly infectious zoonotic disease. C. burnetii has become one of the most important causes of abortion in livestock, which can lead to widespread abortions in these animals. There are very limited studies on the prevalence of C. burnetii infection in cases of animal abortion in Iran. The aim of this study was to investigate the occurrence of C. burnetii in ruminant abortion samples in Iran. Methods Abortion samples from cattle, sheep and goats were collected from different parts of Iran and were tested using Real-time PCR targeting the IS1111 element of C. burnetii. Results In this study, 36 samples (24.7%) of the 146 collected samples were positive for C. burnetii. The prevalence of C. burnetii was 21.3% (20 of 94 samples) in sheep samples. Also, 10 of 46 cattle samples (21.7%) were positive. All six goat abortion samples were positive for C. burnetii. Conclusions The findings of the study demonstrate that C. burnetii plays an important role in domestic ruminant abortions in Iran, suggesting that more attention should be paid to the role of C. burnetii in domestic animal abortions by veterinary organizations. The risk of transmitting the infection to humans due to abortion of animals should also be considered.

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Seroprevalence of Coxiella burnetii among ewes and associated risk factors in Constantine (Northeastern Algeria)

Journal of the Hellenic Veterinary Medical Society ◽

10.12681/jhvms.25100 ◽

2020 ◽

Vol 71 (3) ◽

pp. 2383

Author(s):

S. HIRECHE ◽

A. AGABOU ◽

Ο. BOUAZIZ

Keyword(s):

Risk Factors ◽

Coxiella Burnetii ◽

Q Fever ◽

Elisa Test ◽

Pest Infestation ◽

Domestic Ruminants ◽

North Eastern ◽

Surveillance Programs ◽

Eastern Algeria ◽

Associated Risk Factors

Q fever is a zoonotic disease caused by the rickettsia-like Coxiella burnetii and leads to abortions and decreased reproductive performances in domestic ruminants. A serological survey, using ELISA test, was conducted to assess the prevalence of this infection in 226 ewes belonging to 39 flocks localized in Constantine (North-eastern Algeria). A pretested questionnaire has been submitted to farmers/shepherds to collect information related to relevant risk factors. Results revealed the presence of C. burnetii antibodies in 12.4% (95% CI: 8.08%−16.72%) of individual animals while 35.9% (95% CI: 21.20%−52.82%) of sampled flocks accounted at least one seropositive ewe. Significant causative associations were observed for origin of animals (χ2=14.29, P=0.001), vaccination against enterotoxaemia (χ2=12.12, P=0.002) and pox (χ2=5.30, P=0.025), access to the farm by foreign visitors (χ2=10.87, P=0.004), farmers/shepherds’ visits to other farms (χ2=6.31, P=0.021), disinfection frequency (χ2=7.98, P=0.046), pest infestation within farms (χ2=9.55, P=0.049) and abortion history (χ2=5.54, P=0.029). This recorded prevalence of Coxiella infection would indicate a possible responsibility of this agent in causing abortion and reproductive failures in the tested flocks. Implementing active surveillance programs and further investigations using more accurate analyses and including large samples of more animal species from several provinces are needed to eluci date the real occurrence and dynamics of this infection in the national livestock.

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