Molecular detection of Brucella spp. in ruminant herds in Greece

Katsiolis, Aristomenis; Papanikolaou, Eleni; Stournara, Athanasia; Giakkoupi, Panagiota; Papadogiannakis, E.; Zdragas, Antonis; Giadinis, Nektarios D.; Petridou, Evanthia

doi:10.1007/s11250-022-03175-x

Cited by 6 publications

(5 citation statements)

References 21 publications

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“…Although brucellosis is a notifiable disease in most European countries and most EU Member States are considered brucellosis free [ 4 ], the disease is still endemic, especially in the Mediterranean region [ 1 , 47 ]. Greece, in particular, repeatedly reports high incidences of brucellosis in humans with the main causative agent being B. melitensis [ 6 , 7 , 19 ] and domestic ruminants as the main natural reservoir [ 19 ]. The results of the present study substantiate these findings, although, with one exception, no epidemiological connection between isolates from humans and small ruminants could be drawn from the collected isolates.…”

Section: Discussionmentioning

confidence: 99%

“…Higher resolution can be achieved by using whole-genome sequencing (WGS)-based techniques, such as the analysis of single nucleotide polymorphisms (SNPs) or thousands of target genes in a core genome MLST (cgMLST) approach [ 18 ]. Based on genomic analysis, five major B. melitensis genotypes, the main causative agent of brucellosis in Greece [ 6 , 7 , 19 ], have been identified, which in turn include several subgenotypes [ 20 ]. In addition, these methods allow linkage of human brucellosis cases to Brucella strains isolated from animals or their products [ 11 ].…”

Section: Introductionmentioning

confidence: 99%

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Genotype diversity of brucellosis agents isolated from humans and animals in Greece based on whole-genome sequencing

Brangsch,

Sandalakis,

Babetsa

et al. 2023

BMC Infect Dis

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Background Brucellosis is a zoonotic disease whose causative agent, Brucella spp., is endemic in many countries of the Mediterranean basin, including Greece. Although the occurrence of brucellosis must be reported to the authorities, it is believed that the disease is under-reported in Greece, and knowledge about the genomic diversity of brucellae is lacking. Methods Thus, 44 Brucella isolates, primarily B. melitensis, collected between 1999 and 2009 from humans and small ruminants in Greece were subjected to whole genome sequencing using short-read technology. The raw reads and assembled genomes were used for in silico genotyping based on single nucleotide substitutions and alleles. Further, specific genomic regions encoding putative virulence genes were screened for characteristic nucleotide changes, which arose in different genotype lineages. Results In silico genotyping revealed that the isolates belonged to three of the known sublineages of the East Mediterranean genotype. In addition, a novel subgenotype was identified that was basal to the other East Mediterranean sublineages, comprising two Greek strains. The majority of the isolates can be assumed to be of endemic origin, as they were clustered with strains from the Western Balkans or Turkey, whereas one strain of human origin could be associated with travel to another endemic region, e.g. Portugal. Further, nucleotide substitutions in the housekeeping gene rpoB and virulence-associated genes were detected, which were characteristic of the different subgenotypes. One of the isolates originating from an aborted bovine foetus was identified as B. abortus vaccine strain RB51. Conclusion The results demonstrate the existence of several distinct persistent Brucella sp. foci in Greece. To detect these and for tracing infection chains, extensive sampling initiatives are required.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Genotype diversity of brucellosis agents isolated from humans and animals in Greece based on whole-genome sequencing

Brangsch,

Sandalakis,

Babetsa

et al. 2023

BMC Infect Dis

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“…In Greece, both control and eradication programs against brucellosis of small ruminants are applied, depending on the geographical area of the country [ 11 , 25 ]. Brucellosis was reported significantly more frequently in the continental area of the country and very infrequently in the islands, in line with previous (over 25 years ago) findings of testing people living in the island of Crete, in which antibodies against Brucella spp.…”

Section: Discussionmentioning

confidence: 99%

Zoonotic Problems Reported by Sheep and Goat Farmers and Factors Potentially Contributing to the Occurrence of Brucellosis among Them

Lianou

Petinaki

Michael

et al. 2022

IJERPH

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The study aimed to investigate the occurrence of zoonotic problems reported by dairy small ruminant farmers in Greece and to study potential associations with socio-demographic characteristics of the farmers and management practices applied in the farms. A countrywide investigation was performed in 325 sheep and 119 goat farms in the 13 administrative regions of Greece. The selected farms were visited and interviews were conducted with respective farmers. The occurrence of zoonotic problems in the farmers was recorded. A total biosecurity score (0–6) was devised, based on biosecurity practices followed in farms. Sixty-seven farmers (15.10%, 95% confidence intervals (CI): 12.1–18.7%) reported experiencing a zoonotic problem. Most of the farmers (n = 57) (85.1%, 95% CI: 74.76–91.7%, of those with a zoonotic problem) (12.8%, 95% CI: 10.0–16.3%, of all) reported that the zoonotic problem had been brucellosis. Odds ratio for the occurrence of brucellosis in goat farmers was 1.879 (95% CI: 1.051–3.359) compared to the occurrence of the infection in sheep farmers (p = 0.033). For the outcome ‘occurrence of brucellosis’ in sheep farmers, the application of hand-milking, the availability of a separate lambing area and the presence of cats in the farm emerged as significant (p < 0.01); for the same outcome in goat farmers, only the availability of a separate kidding area emerged as significant (p = 0.001). The mean biosecurity score in farms in the continental area of the country was significantly higher than in the islands: 3.45 ± 0.05 versus 2.76 ± 0.28, respectively (p = 0.006), whilst there was also a significantly higher score in farms, where the farmer reported occurrence of brucellosis: 3.68 ± 0.15 versus 3.34 ± 0.06 in farms, where the farmer did not report such an incident (p = 0.042). In farms, where the above predictors prevail, farmers should be warned of an increased potential risk for human infection and biosecurity measures should be implemented and tightened.

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“…Those samples derived either from positive males to Rose Bengal Test (RBT) and/ or Component Fixation Test (CFT) either from aborted a Parentheses indicate numbers of markers that worked properly in MLVA identification fetuses of sheep, goats, and cattle. Thirty-nine (39) out of the 264 tissue samples, which derived from 30 animals, were found PCR positive using the screening Multiplex PCR detection molecular methodology of Garcia-Yoldi et al [29] for the needs of the Greek Ministry of Rural Development [51] and were utilized for the needs of the present study. The 39 samples originated from various organs and tissues from 30 male and female domestic ruminants, i.e.…”

Section: Sample Collectionsmentioning

confidence: 99%

“…The above 39 samples were found positive by the Multiplex PCR Assay for Brucella spp. [29] in our laboratory [51]. All samples are displayed in detail in Table 1.…”

Section: Sample Collectionsmentioning

confidence: 99%

Brucella spp. distribution, hosting ruminants from Greece, applying various molecular identification techniques

et al. 2022

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Background Brucellosis still remains an endemic disease for both livestock and human in Greece, influencing the primary sector and national economy in general. Although farm animals and particularly ruminants constitute the natural hosts of the disease, transmission to humans is not uncommon, thus representing a serious occupational disease as well. Under this prism, knowledge concerning Brucella species distribution in ruminants is considered a high priority. There are various molecular methodologies for Brucella detection with however differential discriminant capacity. Hence, the aim of this survey was to achieve nationally Brucella epidemiology baseline genotyping data at species and subtype level, as well as to evaluate the pros and cons of different molecular techniques utilized for detection of Brucella species. Thirty-nine tissue samples from 30 domestic ruminants, which were found positive applying a screening PCR, were tested by four different molecular techniques i.e. sequencing of the 16S rRNA, the BP26 and the OMP31 regions, and the MLVA typing panel 1 assay of minisatellite markers. Results Only one haplotype was revealed from the 16S rRNA sequencing analysis, indicating that molecular identification of Brucella bacteria based on this marker might be feasible solely up to genus level. BP26 sequencing analysis and MLVA were in complete agreement detecting both B. melitensis and B. abortus. An interesting exception was observed in 11 samples, of lower quality extracted DNA, in which not all expected MLVA amplicons were produced and identification was based on the remaining ones as well as on BP26. On the contrary OMP31 failed to provide a clear band in any of the examined samples. Conclusions The present study reveals the constant circulation of Brucella bacteria in ruminants throughout Greece. Further, according to our results, BP26 gene represents a very good alternative to MLVA minisatellite assay, particularly in lower quality DNA samples.

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Molecular detection of Brucella spp. in ruminant herds in Greece

Cited by 6 publications

References 21 publications

Genotype diversity of brucellosis agents isolated from humans and animals in Greece based on whole-genome sequencing

Genotype diversity of brucellosis agents isolated from humans and animals in Greece based on whole-genome sequencing

Zoonotic Problems Reported by Sheep and Goat Farmers and Factors Potentially Contributing to the Occurrence of Brucellosis among Them

Brucella spp. distribution, hosting ruminants from Greece, applying various molecular identification techniques

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