In this study, the vaccination coverage, serological sampling and infection rate of sheep and goats were evaluated as predictors for the modeling of human brucellosis in Greece. The human brucellosis disease frequency per local regional unit (RU) varied significantly (RR90) among consecutive years. The notification rate was higher (p < 0.001) in the RUs with implementation of vaccination in sheep and goats (vaccination zone—VZ) with a median of 1.4 (IQR 0.0–3.1) compared with the RUs of the eradication zone (EZ) with a median of 0.0 (IQR 0.0–0.0). In VZ, the increased frequency of human cases was associated with delayed vaccine administration (estimate: 0.14 (0.04; 0.29), p = 0.03) and higher vaccination coverage of the animals (estimate: −0.349 (−0.72; −0.07), p < 0.01). However, the flock sampling rate was highly heterogenous among RUs (IQR 10.56–52.93), and inconsistent within RUs throughout the period of the study 2013–2017 (p = 0.001), limiting the reliable estimation of the infection rate in livestock and the design of an integrated One Health model for human disease.

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 concerningBrucellaspecies distribution in ruminants is considered a high priority. There are various molecular methodologies forBrucelladetection with however differential discriminant capacity. Hence, the aim of this survey was to achieve nationallyBrucellaepidemiology baseline genotyping data at species and subtype level, as well as to evaluate the pros and cons of different molecular techniques utilized for detection ofBrucellaspecies. 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 ofBrucellabacteria based on this marker might be feasible solely up to genus level. BP26 sequencing analysis and MLVA were in complete agreement detecting bothB. melitensisandB. 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 ofBrucellabacteria 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.

The sheep and goats brucellosis program is one of the most important animalhealth programs with significant animal health, public health, and internationaltrade consequences. Several factors are involved in the implementation of thisprogram, while its success depends on the availability of an effective, well-trained,and fully engaged staff. Considering the poor results of sheep and goats brucellosisprogram in Greece mainly related to the limited number of veterinarians workingin the regional veterinary services, the present study aims to approach the numberof the human resources and more precisely of veterinarians that are essential forthe effective and efficient brucellosis control strategy in Greece. Therefore, the dataderived by the performance of the program for the years 2012 to 2015 are analyzed.Finally, economic and political circumstances – external factors that affect thefunctionality of the program as well as all relevant bottlenecks are being discussed.The findings of the present study indicate that approximately 485 veterinarians with485 assistants working for the needs of the brucellosis program would significantlyassist on the complete implementation of the brucellosis control and eradication ofthe disease in Greek sheep and goat holdings.

Background: Zoonotic tuberculosis has been mainly associated with the consumption of bovine dairy products and its control has been prioritized by the Food and Agriculture Organisation. Objective: As such, the aim of this study is to assess whether the polymerase chain reaction (PCR) can be used as an early diagnostic indicator of Mycobacterium bovis infection in bovine dairy farms. Methods: Milk samples (n=78) were collected from all the animals older than 8 years of age, in all (n=4) bovine dairy farms located in a specific agricultural region of Greece, with high prevalence in bovine tuberculosis. Detection of DNA belonging to slow growing members of the genus Mycobacterium was conducted in pooled samples using two PCR assays targeting 16S-rRNA and 65-kDa heat shock protein (hsp65). Randomly selected PCR products were submitted to sequence analysis for confirmation of the specificity of the amplification process. DNA isolation and PCR testing were conducted in compliance with ISO17025 accreditation requirements. Results: The overall percentage of positivity was 47.7%, and ranged among farms from 0% to 76.9%. PCRpositive animals were detected in both farms that were at the time of investigation positive with the tuberculin skin test (TST), whereas the only farm with a negative TST record tested also negatively with PCR. Interestingly, one farm that was negative with TST since 2012 but had a long prior record of high level TST-positivity, tested positively with PCR. Conclusion: In conclusion it can be stated that PCR can be used for the detection of mycobacteria in pooled samples of milk collected from the older animals of a dairy farm, as an early and sensitive diagnostic indicator. This can support TST monitoring for the control of bovine tuberculosis, and improve detection of farms, in which routine monitoring should be intensified. The specific approach offers significant practical benefits that compensate for the additional cost of PCR.