Genotyping Brucella canis isolates using a highly discriminatory multilocus variable-number tandem-repeat analysis (MLVA) assay

Yang, Yi; Wang, Yin; Poulsen, Elizabeth; Ransburgh, Russell; Liu, Xuming; An, Baoyan; Lu, Nanyan; Anderson, Gary A.; Wang, Chengming; Bai, Jianfa

doi:10.1038/s41598-017-01114-7

Cited by 13 publications

(9 citation statements)

References 35 publications

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“…These findings indicate no obvious epidemiological relatedness among these strains and reveal that sporadic cases dominate the incidence of canine brucellosis in China. Previous studies have confirmed that MLVA‐16 genotyping results and epidemiological data are well correlated, with epidemiologically related isolates displaying identical or very closely related genotypes (Allen et al, ; Kilic et al, ; Yang et al, ). The seven genotypes were found in a total of 20 isolates, with each genotype represented by two to six isolates.…”

Section: Discussionmentioning

confidence: 74%

Investigation of the molecular epizootiological characteristics and tracking of the geographical origins ofBrucella canisstrains in China

Liu

Wang

et al. 2019

Transbound Emerg Dis

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Brucellosis is a global pandemic infectious zoonosis. Brucella canis is a rare source of human brucellosis in China, and its public health significance remains under debate. Moreover, data pertaining to the epizootiological characteristics and geographical origin of B. canis on a nationwide scale are limited, and the risk to public safety posed by B. canis infections is unknown. The MLVA (multilocus variable‐number tandem repeat analysis) assay can be helpful to analyse epidemiological correlations among Brucella isolates and to track their geographic origins. To accomplish this task, MLVA‐16 was used to analyse the epidemiological links of 63 isolates obtained from dogs and humans. Sixty‐three B. canis strains were sorted into three large clusters (A, B and C) and 50 different genotypes (GT1–50), and 43 unique genotypes were represented by single isolates, suggesting that these strains had no obvious epidemiological links and that canine brucellosis is predominantly sporadic in China. The other seven shared genotypes (among a total of 20 isolates) were each represented by two to eight isolates, indicating that strains from each shared genotype were epidemiologically correlated. Five of the shared genotypes were from 16 strains obtained from Beijing, indicating that canine brucellosis in Beijing originates from multipoint outbreaks with multiple sources of infection. Based on comprehensive case analysis of clinical B. canis infection, we preliminarily suggest that human B. canis infections are associated with Mycoplasma pneumoniae infection that results in decreased patient immunity. B. canis may have limited epidemiological significance for the healthy population, but it remains a significant threat to the canine breeding industry and to humans who come into close contact with dogs. Based on MLVA‐11 data, B. canis strains were clustered into 16 genotypes and divided into five evolutionary branches; these data confirm that this population covers an extensive geographic area and exhibits characteristics of the origin and evolution of co‐existing introduced and locally native lineages. We believe this study will contribute to strengthening efforts to prevent and control canine brucellosis and to improve public understanding of the health risks posed by B. canis.

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Section: Discussionmentioning

confidence: 74%

Investigation of the molecular epizootiological characteristics and tracking of the geographical origins ofBrucella canisstrains in China

Liu

Wang

et al. 2019

Transbound Emerg Dis

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“…Besides the serological, biochemical and MALDI-TOF MS-based identification tools described above, molecular methods like ribotyping, pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST) or multiple locus variable number of tandem repeats (MLVA) analysis are used for subtyping of Brucella spp. 35 – 37 and for the differentiation of B. canis subclades 38 . As a consequence of the high genetic homology among Brucella species 39 , we performed next-generation sequencing (NGS) analyses for a more sophisticated phylogenetic characterization of the Brucella isolates in our study.…”

Section: Resultsmentioning

confidence: 99%

MALDI-TOF MS and genomic analysis can make the difference in the clarification of canine brucellosis outbreaks

Silva

Brendebach

Grützke

et al. 2020

Sci Rep

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Brucellosis is one of the most common bacterial zoonoses worldwide affecting not only livestock and wildlife but also pets. Canine brucellosis is characterized by reproductive failure in dogs. Human Brucella canis infections are rarely reported but probably underestimated due to insufficient diagnostic surveillance. To improve diagnostics, we investigated dogs in a breeding kennel that showed clinical manifestations of brucellosis and revealed positive blood cultures. As an alternative to the time-consuming and hazardous classical identification procedures, a newly developed species-specific intact-cell matrix-assisted laser desorption/ionization–time of flight mass spectrometry analysis was applied, which allowed for rapid identification of B. canis and differentiation from closely related B. suis biovar 1. High-throughput sequencing and comparative genomics using single nucleotide polymorphism analysis clustered our isolates together with canine and human strains from various Central and South American countries in a distinct sub-lineage. Hence, molecular epidemiology clearly defined the outbreak cluster and demonstrated the endemic situation in South America. Our study illustrates that MALDI-TOF MS analysis using a validated in-house reference database facilitates rapid B. canis identification at species level. Additional whole genome sequencing provides more detailed outbreak information and leads to a deeper understanding of the epidemiology of canine brucellosis.

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“…This was also true for the subset of sexually intact purebred adult dogs of breeding age (n = 1,397) identified as undergoing routine prebreeding serologic screening; 958 (68.6%) of these dogs were female and 439 (31.4%) were male. Investigators of a study 17 on B canis genotypes in samples submitted to the state veterinary diagnostic laboratory in Kansas also identified more samples from female (87/133 [65%]) than male dogs. Yet in the present study, 4 of the 7 dogs with diskospondylitis were males, 2 were females, and the sex of 1 was not reported.…”

Section: Discussionmentioning

confidence: 99%

“…41 The variety of DNA extraction methods, primers, and amplification techniques used to detect B canis in dogs make direct comparisons among study results difficult. 17,40,41 At present, it appears that commonly used commercial kits for DNA extraction from canine blood samples are not, by themselves, sufficiently sensitive to recommend replacing blood culture for diagnosis of B canis in dogs. 40,41 Results depend on the quality of the sample submitted as well as the methodology used at the diagnostic laboratory and experience of personnel.…”

Section: Discussionmentioning

confidence: 99%

Investigation and characterization of Brucella canis infections in pet-quality dogs and associated human exposures during a 2007–2016 outbreak in Michigan

Johnson¹,

Carter²,

Dunn³

et al. 2018

javma

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OBJECTIVE To estimate Brucella canis seropositivity rates for purebred dogs being bred by noncommercial breeders, describe epidemiological findings in infected commercial dog-production facilities, and characterize B canis infection in pet dogs and the risk to human health. DESIGN Retrospective descriptive study. SAMPLE 2,799 canine specimens submitted to the Michigan State University Veterinary Diagnostic Laboratory for B canis testing and records of B canis reports provided to the Michigan Department of Agriculture and Rural Development from 2007 through 2016. PROCEDURES Results of B canis laboratory tests and epidemiological findings for reported cases of B canis were reviewed and summarized. Federal and state public health officials were interviewed regarding human B canis infection. State veterinarians were interviewed regarding canine brucellosis reporting and control procedures. RESULTS Estimated B canis seropositivity was 0.4% among purebred Michigan dogs owned by noncommercial breeders. Infection was confirmed in dogs from 17 commercial dog-production facilities, 3 shelters, and 1 rescue agency. Estimated infection prevalence in production facilities ranged from 2 of 22 (9%) to 5 of 6 (83%). Transfer of infected dogs involved 22 Michigan counties and 11 states. Seven of 20 privately owned infected dogs had diskospondylitis; I also had uveitis. Fifty-three veterinary hospital or diagnostic laboratory personnel had inadvertent exposure to the pathogen. Brucella canis was isolated from 1 commercial production facility owner. CONCLUSIONS AND CLINICAL RELEVANCE B canis was uncommon in purebred dogs being bred by noncommercial breeders but endemic in Michigan commercial facilities producing dogs destined to become household pets. Infected pet dogs caused human B canis exposure, and several pet dogs had debilitating disease not associated with the reproductive system.

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Genotyping Brucella canis isolates using a highly discriminatory multilocus variable-number tandem-repeat analysis (MLVA) assay

Cited by 13 publications

References 35 publications

Investigation of the molecular epizootiological characteristics and tracking of the geographical origins ofBrucella canisstrains in China

Investigation of the molecular epizootiological characteristics and tracking of the geographical origins ofBrucella canisstrains in China

MALDI-TOF MS and genomic analysis can make the difference in the clarification of canine brucellosis outbreaks

Investigation and characterization of Brucella canis infections in pet-quality dogs and associated human exposures during a 2007–2016 outbreak in Michigan

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