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

2015 ◽  
Vol 81 (20) ◽  
pp. 7253-7260 ◽  
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.

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

2021 ◽  
Vol 10 ◽  
Author(s):  
Sara Tomaiuolo ◽  
Samira Boarbi ◽  
Tiziano Fancello ◽  
Patrick Michel ◽  
Damien Desqueper ◽  
...  
Keyword(s):  
Coxiella Burnetii ◽  
Small Ruminant ◽  
Q Fever ◽  
Variable Number Tandem Repeat ◽  
Primary Source ◽  
Variable Number ◽  
Human Infection ◽  
Apparent Prevalence ◽  
Domestic Ruminants ◽  
Contamination Routes

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.

scholarly journals Four-Year Evaluation of the Effect of Vaccination against Coxiella burnetii on Reduction of Animal Infection and Environmental Contamination in a Naturally Infected Dairy Sheep Flock

2011 ◽  
Vol 77 (20) ◽  
pp. 7405-7407 ◽  
Author(s):  
Ianire Astobiza ◽  
Jesús F. Barandika ◽  
Francisco Ruiz-Fons ◽  
Ana Hurtado ◽  
Inés Povedano ◽  
...  
Keyword(s):  
Phase I ◽  
Coxiella Burnetii ◽  
Environmental Contamination ◽  
Sheep Flock ◽  
Dairy Sheep ◽  
Content Type ◽  
Animal Infection

ABSTRACTVaccination is considered one of the best options for controllingCoxiella burnetiiinfection in livestock. The efficacy of a phase I vaccine was investigated over 4 years in a sheep flock with confirmedC. burnetiiinfection. Shedding was not detected in ewes and yearlings in the last 2 years, butC. burnetiistill persisted in the environment.

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

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.

scholarly journals 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

2016 ◽  
Vol 4 (2) ◽  
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.

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

Pathogens ◽  
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.

scholarly journals Coxiella burnetii Circulation in a Naturally Infected Flock of Sheep: Individual Follow-Up of Antibodies in Serum and Milk

2017 ◽  
Vol 83 (13) ◽  
Author(s):  
A. Joulié ◽  
E. Rousset ◽  
P. Gasqui ◽  
E. Lepetitcolin ◽  
A. Leblond ◽  
...  
Keyword(s):  
Coxiella Burnetii ◽  
Field Data ◽  
Q Fever ◽  
Maternal Antibodies ◽  
Content Type ◽  
Milk Samples ◽  
Ewe Lambs ◽  
Antibody Levels ◽  
Over Time

ABSTRACT The control of Q fever, a zoonotic disease caused by the Coxiella burnetii bacterium, remains a scientific challenge. Domestic ruminants are considered the main reservoir, shedding C. burnetii essentially through parturition products during abortion or birth. Sheep are particularly frequently associated with human outbreaks, but there are insufficient field data to fully understand disease dynamics and to instigate efficient control measures. A longitudinal follow-up study of a naturally infected sheep flock was performed (i) to investigate relationships between seropositivity and bacterial shedding in the vaginal mucus, (ii) to describe the kinetics of antibodies, including responses to vaccination, (iii) to monitor maternal antibodies in ewe lambs, and (iv) to compare serological results for milk and serum samples. For 8 months, we collected blood samples every 3 weeks from 11 aborting and 26 nonaborting dairy ewes, 20 nonaborting suckler ewes, and 9 ewe lambs. Individual milk samples were also obtained from lactating females. All serum and milk samples were tested by enzyme-linked immunosorbent assay (ELISA), whereas vaginal swabs were tested by quantitative PCR. We found that some dairy females did not seroconvert despite shedding C. burnetii in their vaginal mucus. Overall, antibody levels in adult females were found to remain stable over time, with exceptions during the mating and lambing periods. Maternal antibodies decreased during the first month after birth. Interestingly, antibody levels in milk were correlated with those in serum. This study provides valuable field data that will help improve Q fever surveillance and within-flock management measures. IMPORTANCE Field data are necessary to improve the surveillance, diagnosis, and sanitary management of Q fever in livestock. Here, we provide extensive serological data obtained from serum and milk samples from infected and vaccinated ewes belonging to a naturally infected flock of sheep. We show that antibody levels are stable over time and seropositivity and vaginal shedding are not clearly correlated, whereas antibody levels in milk are strongly correlated with those in serum. Accordingly, we find that antibody levels in bulk tank milk are consistent with the variations observed in the serum of dairy females over time. We report the existence of maternal antibody transmission to ewe lambs and we show that the presence of maternal antibodies at birth does not prevent the development of a serological response to vaccination at the age of 4 months. Finally, we report that adult ewes generally seroconvert after vaccination, including during pregnancy.

scholarly journals Correlating Genotyping Data of Coxiella burnetii with Genomic Groups

Pathogens ◽  
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.

scholarly journals A Q Fever Outbreak with a High Rate of Abortions at a Dairy Goat Farm:Coxiella burnetiiShedding, Environmental Contamination, and Viability

2018 ◽  
Vol 84 (20) ◽  
Author(s):  
Raquel Álvarez-Alonso ◽  
Mikel Basterretxea ◽  
Jesús F. Barandika ◽  
Ana Hurtado ◽  
Jasone Idiazabal ◽  
...  
Keyword(s):  
Coxiella Burnetii ◽  
Environmental Contamination ◽  
Q Fever ◽  
Vero Cells ◽  
Control Measures ◽  
High Rate ◽  
Dairy Goat ◽  
Content Type ◽  
Infection Dynamics ◽  
Low Incidence

ABSTRACTThis study describes a Q fever outbreak in a herd of 77 Alpine goats which suffered a high rate of abortions (81% [58/72]) in January 2017 and presents the results of monitoring the contamination and viability ofCoxiella burnetiiin the farm environment several months after the outbreak. Over the course of 7 months, we studied bacterial shedding by 35 dams with abortions to monitorC. burnetiiinfection dynamics and the duration of excretion. The highest bacterial shedding load was observed in vaginal mucus, followed by in feces and in milk. Conversely, the duration ofC. burnetiishedding was longer through feces (5 months after abortion) than milk (3 months).C. burnetiiDNA was detected throughout the study in aerosol samples periodically collected indoors and outdoors from the animal premises. Mouse inoculation and culture in Vero cells demonstrated the presence of viable isolates in dust collected from different surfaces inside the animal facilities during the period of time with the highest number of abortions but not in dust collected 2, 3, and 4 months after the last parturition. Some workers and visitors were affected by Q fever, with attack rates of 78% (7/9) and 31% (4/13), respectively. Affected people mostly showed fever and seroconversion, along with myalgia and arthralgia in two patients and pneumonia in the index case. The genotype identified in animal and environmental samples (SNP1/MST13) turned out to be very aggressive in goats but caused only moderate symptoms in people. After the diagnosis of abortion by Q fever in goats, several control measures were implemented at the farm to prevent contamination inside and outside the animal facilities.IMPORTANCEThis work describes a 7-month follow-up of the excretion by different routes ofCoxiella burnetiigenotype SNP1/MST13 in a herd of goats that suffered high rate of abortions (81%), generating high environmental contamination. Some of the workers and visitors who accessed the farm were infected, with fever as the main symptom but a low incidence of pneumonia. The detected strain (SNP1/MST13 genotype) turned out to be very aggressive in goats. The viability ofC. burnetiiwas demonstrated in the environment of the farm at the time of abortions, but 2 months after the last parturition, no viable bacteria were detected. These results highlighted the importance of implementing good biosafety measures at farms and avoiding the entrance of visitors to farms several months after the end of the kidding period.

scholarly journals Potentially Human-Pathogenic Escherichia coli O26 in Norwegian Sheep Flocks

2011 ◽  
Vol 77 (14) ◽  
pp. 4949-4958 ◽  
Author(s):  
C. Sekse ◽  
M. Sunde ◽  
B.-A. Lindstedt ◽  
P. Hopp ◽  
T. Bruheim ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Variable Number Tandem Repeat ◽  
Variable Number ◽  
Human Pathogens ◽  
Content Type ◽  
Representative Sampling ◽  
E Coli ◽  
Pathogenic Escherichia Coli ◽  
Close Relationship ◽  
Large Survey

ABSTRACTA national survey ofEscherichia coliO26 in Norwegian sheep flocks was conducted, using fecal samples to determine the prevalence. In total, 491 flocks were tested, andE. coliO26 was detected in 17.9% of the flocks. One hundred forty-twoE. coliO26 isolates were examined for flagellar antigens (H typing) and four virulence genes, includingstxandeae, to identify possible Shiga toxin-producingE. coli(STEC) and enteropathogenicE. coli(EPEC). Most isolates (129 out of 142) were identified asE. coliO26:H11. They possessedeaeand may have potential as human pathogens, although only a small fraction were identified as STEC O26:H11, giving a prevalence in sheep flocks of only 0.8%. Correspondingly, the sheep flock prevalence of atypical EPEC (aEPEC) O26:H11 was surprisingly high (15.9%). The genetic relationship between theE. coliO26:H11 isolates was investigated by pulsed-field gel electrophoresis (PFGE) and multilocus variable number tandem repeat analysis (MLVA), identifying 63 distinct PFGE profiles and 22 MLVA profiles. Although the MLVA protocol was less discriminatory than PFGE and a few cases of disagreement were observed, comparison by partition mapping showed an overall good accordance between the two methods. A close relationship between a few isolates of aEPEC O26:H11 and STEC O26:H11 was identified, but all theE. coliO26:H11 isolates should be considered potentially pathogenic to humans. The present study consisted of a representative sampling of sheep flocks from all parts of Norway. This is the first large survey of sheep flocks focusing onE. coliO26 in general, including results of STEC, aEPEC, and nonpathogenic isolates.

scholarly journals Coxiella burnetii Alters Cyclic AMP-Dependent Protein Kinase Signaling during Growth in Macrophages

2012 ◽  
Vol 80 (6) ◽  
pp. 1980-1986 ◽  
Author(s):  
Laura J. MacDonald ◽  
Richard C. Kurten ◽  
Daniel E. Voth
Keyword(s):  
Protein Kinase ◽  
Cyclic Amp ◽  
Coxiella Burnetii ◽  
Alveolar Macrophages ◽  
Q Fever ◽  
Parasitophorous Vacuole ◽  
Dependent Protein Kinase ◽  
Content Type ◽  
Bacterial Agent ◽  
Dependent Protein

ABSTRACTCoxiella burnetiiis the bacterial agent of human Q fever, an acute, flu-like illness that can present as chronic endocarditis in immunocompromised individuals. Following aerosol-mediated transmission,C. burnetiireplicates in alveolar macrophages in a unique phagolysosome-like parasitophorous vacuole (PV) required for survival. The mechanisms ofC. burnetiiintracellular survival are poorly defined and a recent Q fever outbreak in the Netherlands emphasizes the need for better understanding this unique host-pathogen interaction. We recently demonstrated that inhibition of host cyclic AMP-dependent protein kinase (PKA) activity negatively impacts PV formation. In the current study, we confirmed PKA involvement in PV biogenesis and probed the role of PKA signaling duringC. burnetiiinfection of macrophages. Using PKA-specific inhibitors, we found the kinase was needed for biogenesis of prototypical PV andC. burnetiireplication. PKA and downstream targets were differentially phosphorylated throughout infection, suggesting prolonged regulation of the pathway. Importantly, the pathogen actively triggered PKA activation, which was also required for PV formation by virulentC. burnetiiisolates during infection of primary human alveolar macrophages. A subset of PKA-specific substrates were differentially phosphorylated duringC. burnetiiinfection, suggesting the pathogen uses PKA signaling to control distinct host cell responses. Collectively, the current results suggest a versatile role for PKA inC. burnetiiinfection and indicate virulent organisms usurp host kinase cascades for efficient intracellular growth.

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