Comparing the genetic typing methods for effective surveillance and rabies control in Georgia

A full nucleoprotein gene sequencing of 68 isolates collected from passive rabies surveillance system in Georgia between 2015 and 2016 identified two distinct dog rabies phylogroups, GEO_V1 and GEO_V2, which both belonged to the cosmopolitan dog clade. GEO_V1 was found throughout the country and was further divided into four sub-phylogroups that overlapped geographically; GEO_V2 was found in the southeast region and was closely related to dog rabies in Azerbaijan. A sequence analysis of the full N gene, partial nucleoprotein gene of N-terminal and C-terminal, and the amplicon sequences of pan-lyssavirus RT-qPCR LN34 showed that all four sequencing approaches provided clear genetic typing results of canine rabies and could further differentiate GEO_V1 and GEO_V2. The phylogenetic analysis results vary and were affected by the length of the sequences used. Amplicon sequencing of the LN34 assay positive samples provided a rapid and cost-effective method for rabies genetic typing, which is important for improving rabies surveillance and canine rabies eradication globally

Avian Influenza Virus Surveillance in Wild Birds in Georgia: 2009-2011

The Caucasus, at the border of Europe and Asia, is important for migration and over-wintering of wild waterbirds. Three flyways, the Central Asian, East Africa-West Asia, and Mediterranean/Black Sea flyways, converge in the Caucasus region. Thus, the Caucasus region might act as a migratory bridge for influenza virus transmission when birds aggregate in high concentrations in the post-breeding, migrating and overwintering periods. Since August 2009, we have established a surveillance network for influenza viruses in wild birds, using five sample areas geographically spread throughout suitable habitats in both eastern and western Georgia. We took paired tracheal and cloacal swabs and fresh feces samples. We collected 8343 swabs from 76 species belonging to 17 families in 11 orders of birds, of which 84 were real-time RT-PCR positive for avian influenza virus (AIV). No highly pathogenic AIV (HPAIV) H5 or H7 viruses were detected. The overall AIV prevalence was 1.6%. We observed peak prevalence in large gulls during the autumn migration (5.3-9.8%), but peak prevalence in Black-headed Gulls in spring (4.2-13%). In ducks, we observed increased AIV prevalence during the autumn post-moult aggregations and migration stop-over period (6.3%) but at lower levels to those observed in other more northerly post-moult areas in Eurasia. We observed another prevalence peak in the overwintering period (0.14-5.9%). Serological and virological monitoring of a breeding colony of Armenian Gulls showed that adult birds were seropositive on arrival at the breeding colony, but juveniles remained serologically and virologically negative for AIV throughout their time on the breeding grounds, in contrast to gull AIV data from other geographic regions. We show that close phylogenetic relatives of viruses isolated in Georgia are sourced from a wide geographic area throughout Western and Central Eurasia, and from areas that are represented by multiple different flyways, likely linking different host sub-populations

Implementation of the One Health approach to fight arbovirus infections in the Mediterranean and Black Sea Region: Assessing integrated surveillance in Serbia, Tunisia and Georgia

Background In the Mediterranean and Black Sea Region, arbovirus infections are emerging infectious diseases. Their surveillance can benefit from one health inter-sectoral collaboration; however, no standardized methodology exists to study One Health surveillance. Methods We designed a situation analysis study to document how integration of laboratory/clinical human, animal and entomological surveillance of arboviruses was being implemented in the Region. We applied a framework designed to assess three levels of integration: policy/institutional, data collection/data analysis and dissemination. We tested the use of Business Process Modelling Notation (BPMN) to graphically present evidence of inter-sectoral integration. Results Serbia, Tunisia and Georgia participated in the study. West Nile Virus surveillance was analysed in Serbia and Tunisia, Crimea-Congo Haemorrhagic Fever surveillance in Georgia. Our framework enabled a standardized analysis of One Health surveillance integration, and BPMN was easily understandable and conducive to detailed discussions among different actors/institutions. In all countries, we observed integration across sectors and levels except in data collection and data analysis. Data collection was interoperable only in Georgia without integrated analysis. In all countries, surveillance was mainly oriented towards outbreak response, triggered by an index human case. Discussion The three surveillance systems we observed prove that integrated surveillance can be operationalized with a diverse spectrum of options. However, in all countries, the integrated use of data for early warning and inter-sectoral priority setting is pioneeristic. We also noted that early warning before human case occurrence is recurrently not operationally prioritized

Regional study of anthrax foci

ObjectiveThe purpose of this study was to describe anthrax foci alongthe Georgia-Azerbaijan border and to describe control measures inidentified areas.IntroductionAnthrax is endemic in the South Caucasus region. There is alack of understanding of the regional epidemiology of the causativepathogen,Bacillus anthracis, and the trans-boundary factors relatedto its persistence.MethodsTo increase the local and regional understanding of anthraxecology, ecological risk factors, and the genetic relationships anddistribution among Georgian and AzerbaijaniB. anthracisstrains, aregional study of the ecology of anthrax foci was conducted in Georgiaand Azerbaijan. Six regions in Georgia (that border Azerbaijan)were selected for environmental sampling based on historical data.Soil samples were collected in Lagodekhi and Sagarejo and testedat the Laboratory of the Ministry of Agriculture using standardbacteriological and molecular biology methods.ResultsA total of 185 soil samples were collected. Bacteriological testsrevealed four positive samples from Kakheti (two from Lagodekhi,Gelati; two from Dedoplistskaro), from which, cultures were isolatedand confirmed by PCR. Georgian scientists continue collecting andtesting soil samples. After sample collection and bacteriologicaltesting is completed, the molecular characteristics of the pathogenwill be examined.ConclusionsThis study will assist in the formulation of targeted public healthinterventions aimed at increasing knowledge of the disease withinspecific demographics. Public health interventions can focus onlivestock surveillance and control in identified areas

Regional study of anthrax foci

ObjectiveThe purpose of this study was to describe anthrax foci alongthe Georgia-Azerbaijan border and to describe control measures inidentified areas.IntroductionAnthrax is endemic in the South Caucasus region. There is alack of understanding of the regional epidemiology of the causativepathogen,Bacillus anthracis, and the trans-boundary factors relatedto its persistence.MethodsTo increase the local and regional understanding of anthraxecology, ecological risk factors, and the genetic relationships anddistribution among Georgian and AzerbaijaniB. anthracisstrains, aregional study of the ecology of anthrax foci was conducted in Georgiaand Azerbaijan. Six regions in Georgia (that border Azerbaijan)were selected for environmental sampling based on historical data.Soil samples were collected in Lagodekhi and Sagarejo and testedat the Laboratory of the Ministry of Agriculture using standardbacteriological and molecular biology methods.ResultsA total of 185 soil samples were collected. Bacteriological testsrevealed four positive samples from Kakheti (two from Lagodekhi,Gelati; two from Dedoplistskaro), from which, cultures were isolatedand confirmed by PCR. Georgian scientists continue collecting andtesting soil samples. After sample collection and bacteriologicaltesting is completed, the molecular characteristics of the pathogenwill be examined.ConclusionsThis study will assist in the formulation of targeted public healthinterventions aimed at increasing knowledge of the disease withinspecific demographics. Public health interventions can focus onlivestock surveillance and control in identified areas

Intra-epidemic genome variation in highly pathogenic African swine fever virus (ASFV) from the country of Georgia

Abstract Background African swine fever virus (ASFV) causes an acute hemorrhagic infection in suids with a mortality rate of up to 100%. No vaccine is available and the potential for catastrophic disease in Europe remains elevated due to the ongoing ASF epidemic in Russia and Baltic countries. To date, intra-epidemic whole-genome variation for ASFV has not been reported. To provide a more comprehensive baseline for genetic variation early in the ASF outbreak, we sequenced two Georgian ASFV samples, G-2008/1 and G-2008/2, derived from domestic porcine blood collected in 2008. Methods Genomic DNA was extracted directly from low-volume ASFV PCR-positive porcine blood samples and subjected to next generation sequencing on the Illumina Miseq platform. De novo and mapped sequence assemblies were performed using CLCBio software. Genomic illustrations, sequence alignments and assembly figures were generated using Geneious v10.2.4. Sequence repeat architecture was analyzed using DNASTAR GeneQuest 14.1.0. Results The G-2008/1 and G-2008/2 genomes were distinguished from each other by coding changes in seven genes, including MGF 110-1 L, X69R, MGF 505-10R, EP364R, H233R, E199L, and MGF 360-21R in addition to eight homopolymer tract variations. The 2008/2 genome possessed a novel allele state at a previously undescribed intergenic repeat locus between genes C315R and C147L. The C315R/C147L locus represents the earliest observed variable repeat sequence polymorphism reported among isolates from this epidemic. No sequence variation was observed in conventional ASFV subtyping markers. The two genomes exhibited complete collinearity and identical gene content with the Georgia 2007/1 reference genome. Approximately 56 unique homopolymer A/T-tract variations were identified that were unique to the Georgia 2007/1 genome. In both 2008 genomes, within-sample sequence read heterogeneity was evident at six homopolymeric G/C-tracts confined to the known hypervariable ~ 7 kb region in the left terminal region of the genome. Conclusions This is the first intra-epidemic comparative genomic analysis reported for ASFV and provides insight into the intra-epidemic microevolution of ASFV. The genomes reported here, in addition to the G-2007/1 genome, provide an early baseline for future genome-level comparisons and epidemiological tracing efforts

Seroprevalence of Brucellosis in Livestock within Three Endemic Regions of the Country of Georgia

Brucellosis is the one of most common livestock zoonoses in Georgia, resulting in significant economic losses. Livestock were sampled in three regions of Georgia (Kakheti, Kvemo Kartli, Imereti). Districts that historically reported high numbers of brucellosis related morbidity were selected for serological, bacteriological and molecular surveys. Surveying efforts yielded samples from 10,819 large and small ruminants. In total, 735 serological tests were positive on Rose Bengal and 33 bacterial isolates were recovered and identified as Brucella melitensis or Brucella abortus by microbiology and AMOS-PCR. A Bayesian framework was implemented to estimate the true prevalence of the disease given an imperfect diagnostic test. Regional posterior median true prevalence estimates ranged from 2.7% (95% CI: 1.4, 7.2) in Kvemo Kartli, 0.8% (95% CI: 0.0, 3.6) in Kakheti, to an estimate of 0.6% (95% CI: 0.0, 2.9) in Imereti. Accurate and efficient surveillance of brucellosis is not only of economic value, but also informs efforts to reduce the disease impact on the human population

Map of Georgia.

<p>The study areas in Georgia are shown in yellow hatch, the country boundaries in red, and the main geographic features as an altitude relief.</p