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Canine monocytic ehrlichiosis (CME) is a widespread, tick-borne, canine disease, caused by an obligate intracellular bacterium, Ehrlichia canis. The main vector, a brown-dog tick, Rhipicephalus sanguineus, is widely distributed, especially in areas with tropic, subtropic, or Mediterranean climates (Central and South America, Eastern and Western Asia, Africa, Australia and Southern Europe). The study performed in 2012, by Stefanovska et al., determined a seroprevalence of 18.7% of E. canis among the Macedonian dog population. Up to date, the presence of E. canis, using molecular diagnostic methods, has not been investigated in Macedonia. Therefore, this study aimed to confirm the presence of E. canis, in the pet-dog population on the territory of the city of Skopje, North Macedonia, using a highly sensitive multiplex Real-Time PCR method (qPCR). Whole blood samples from 80 dogs of different breeds and ages, with clinical symptoms of CME and positive serology result for the presence of antibodies against E.canis, were collected for analyses. Out of 80 dogs, 36 (45%) were found as positive. The present work reports the first molecular detection of E. canis in pet dogs on the territory of the city of Skopje, Macedonia.

Aim of this study was to characterize pigeon variant of Newcastle disease virus (NDV) isolated from backyard poultry using classical and molecular methods. In standard hemagglutination inhibition (HI) test both polyclonal NDV antiserum and monoclonal antibodies 161/617 specific for pigeon variants of NDV showed inhibition of heamagglutination of the isolated virus. Intracerebral pathogenicity index (ICPI) has shown that the isolate is mesogenic virus (ICPI = 0.81). One-step RT-qPCR for detection of M gene was performed indicating a presence of NDV and RT-qPCR for discrimination between lentogenic and velogenic strains based on F gene was also performed indicating a presence of virulent NDV. A portion of the F gene was amplified and sequenced for determination of virulence and phylogenetic characterization. The F protein cleavage site sequence of the isolate had multiple basic amino acids at residues 112–116 and a phenyl alanine at residue 117 (112RRQKR*F117) which is typical for velogenic strains. The nucleotide sequence of 374 bp was aligned to begin at nt 47 and finish at 420 immediately after the cleavage site and compared with other reference strains from the region and worldwide. In the phylogenetic tree, the isolate clustered into genotype VIb, typical for PPMV-1. This strain is phylogenetically very similar to other PPMV-1 isolated from pigeons in Macedonia. Poultry infected with PPMV-1 can spread the virus in the absence of clinical signs, thus PPMV-1’s are constant threat to domestic poultry. This is the first report of evidenced spillover of PPMV-1 into poultry in Macedonia.

The aim of the study was to determine the presence and distribution or to confirm the absence of avian influenca viruses in farmed poultry in the poultry production systems 1 and 2 from the eight statistical regions in R. Macedonia. Total number of 1215 cloacal swabs from poultry were sampled. Each sample was processed and analysed by both molecular (RRT-PCR) and classical virology methods (virus isolation and identification). All samples gave negative result for presence of avian influenza viruses. Commercial poultry production systems have biosecurity measures preventing the entry of pathogens i.e avian influenza viruses, therefore resulting with no circulation of these viruses in the sampled farmed poultry flocks.

Maedi-Visna in sheep and CAEV in goats are diseases caused by lentiviruses with very similar antigenic characteristics. This diseases are very important for the animal husbandry because of the economical losses they cause (reduced milk yield, lower body weight, deaths, therapy expenses). The results in this article, represents the first serological confirmation for the existence of this diseases in Macedonia. Investigations showed high percent of seroprevalence in the examined herds. The seroprevalence ranges from 60.3% for Maedi-Visna to 55.8% for CAEV. Despite the presence of clinical signs of the disease, the mortality is low, witch is related to the presence of the causal agent in the field i.e. the endemic character of this diseases.

Bluetongue virus serotype 4 (BTV‐4) was confirmed in sheep in North Macedonia in July 2020. The full genome of this BTV‐4 strain (MKD2020/06) was shown to be most closely related (99.74% nt identity) to the Greek GRE2014/08 and the Hungarian HUN1014 strains, indicating the re‐emergence of this BTV serotype in the Balkan region since it was last reported in 2017. [ABSTRACT FROM AUTHOR]

Aim: To analyze the impact of measures undertaken to control brucellosis in the Republic of Macedonia on demographic, epidemiological and clinical characteristics of the disease in humans. Methods: Demographic, epidemiological and clinical characteristics of 340 patients with brucellosis were retrospectively analyzed. Patients were treated at the University Clinic of Infectious Diseases in Skopje during three time periods: 1989-1990 when no national strategy for disease control existed, 2000-2001 when some activities for control were performed and 2011-2014 when an organized and financially supported strategy was accomplished. Results: A drastic decrease of the number of cases of human brucellosis during the last period when compared to the first two ones (3.9 and 3.3 times lower, respectively) was evident. Patients from the third period were significantly older, with median age 41 years (range 4-73) vs. 31 years (3-77) and 34 years (1-74), respectively (p = .013). There were more males (86.6 vs. 73.6% and 63.2%, respectively, p = .002) and the disease was more often acquired professionally (80.6 vs. 60.1% and 59.2%, respectively, p = .006). Fewer patients from the third period had elevated body temperature either as a symptom (50.7 vs. 73.6% and 69.6%, respectively, p = .003), or as a sign (35.8 vs. 55.4% and 56.8%, respectively, p = .011) and less often exhibited focal forms (40.3 vs. 56.8% and 64.0%, respectively, p = .007). Conclusion: The national strategy for brucellosis control, which included vaccination of small ruminants, resulted in a decrease of human brucellosis cases in the Republic of Macedonia and there was a tendency to milder disease manifestations. [ABSTRACT FROM AUTHOR]

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Aim of this study was to characterize pigeon variant of Newcastle disease virus (NDV) isolated from backyard poultry using classical and molecular methods. In standard hemagglutination inhibition (HI) test both polyclonal NDV antiserum and monoclonal antibodies 161/617 specific for pigeon variants of NDV showed inhibition of heamagglutination of the isolated virus. Intracerebral pathogenicity index (ICPI) has shown that the isolate is mesogenic virus (ICPI = 0.81). One-step RT-qPCR for detection of M gene was performed indicating a presence of NDV and RT-qPCR for discrimination between lentogenic and velogenic strains based on F gene was also performed indicating a presence of virulent NDV. A portion of the F gene was amplified and sequenced for determination of virulence and phylogenetic characterization. The F protein cleavage site sequence of the isolate had multiple basic amino acids at residues 112-116 and a phenyl alanine at residue 117 (112RRQKR*F117) which is typical for velogenic strains. The nucleotide sequence of 374 bp was aligned to begin at nt 47 and finish at 420 immediately after the cleavage site and compared with other reference strains from the region and worldwide. In the phylogenetic tree, the isolate clustered into genotype VIb, typical for PPMV-1. This strain is phylogenetically very similar to other PPMV-1 isolated from pigeons in Macedonia. Poultry infected with PPMV-1 can spread the virus in the absence of clinical signs, thus PPMV-1's are constant threat to domestic poultry. This is the first report of evidenced spillover of PPMV-1 into poultry in Macedonia. [ABSTRACT FROM AUTHOR]

Lumpy skin disease virus (LSDV) causes a disease of economic importance affecting cattle. Its global epidemiology is complex due to the combination of vectorborne and anthropogenic spread, the circulation of vaccine-like recombinants, and the use of vaccines. The slow molecular evolution of its DNA genome limits the utility of genetic variation for accurate tracing based on evolutionary analyses, but this limitation has not yet been formally assessed. Furthermore, until present, whole genome sequencing in affected areas has remained patchy. This study combines the first fine-grained sampling of LSDV whole genomes from a time-constrained (2015-2017) southeastern European (SEE) LSDV outbreak, which we analyze along with curated public genomes to investigate the global and regional viral dispersal dynamics. First, haplotype networks visualizing the limited genetic variability associated with the SEE LSDV outbreak show intense intermixing between countries. We also assess at which spatial scale a correlation between genetic and geographic distances can be detected for LSDV. On a global scale, we show the importance of accounting for recombination events that can impact phylogenetic and phylogeographic reconstructions. Following the assessment of the temporal signal in the recombination-free alignment, our time-scaled continuous phylogeographic analysis of Kenya-like and recent wild-type viruses confirms the origin and global dissemination history of LSDV. Our analyses highlight the importance of careful selection and application of phylodynamic approaches to DNA viruses, as well as the importance of whole genome sampling in endemic and outbreak areas to improve our understanding of the evolution, epidemiology, and transmission dynamics of DNA viruses. [ABSTRACT FROM AUTHOR]

Canine monocytic ehrlichiosis (CME) is a widespread, tick-borne, canine disease, caused by an obligate intracellular bacterium, Ehrlichia canis. The main vector, a brown-dog tick, Rhipicephalus sanguineus, is widely distributed, especially in areas with tropic, subtropic, or Mediterranean climates (Central and South America, Eastern and Western Asia, Africa, Australia and Southern Europe). The study performed in 2012, by Stefanovska et al., determined a seroprevalence of 18.7% of E. canis among the Macedonian dog population. Up to date, the presence of E. canis, using molecular diagnostic methods, has not been investigated in Macedonia. Therefore, this study aimed to confirm the presence of E. canis, in the pet-dog population on the territory of the city of Skopje, North Macedonia, using a highly sensitive multiplex Real-Time PCR method (qPCR). Whole blood samples from 80 dogs of different breeds and ages, with clinical symptoms of CME and positive serology result for the presence of antibodies against E.canis, were collected for analyses. Out of 80 dogs, 36 (45%) were found as positive. The present work reports the first molecular detection of E. canis in pet dogs on the territory of the city of Skopje, Macedonia. [ABSTRACT FROM AUTHOR]

Until 2009 no data are available for the circulation of avian influenza viruses (AIV) in backyard poultry in Republic of Macedonia. In 2009 virological and molecular surveillance was conducted in backyard poultry in the risk areas of the country according the National Annual Program for AI and Commission Decision 2007/268/EC. A total number of 2151 samples were analyzed. Two thousand and twenty cloacal swabs (sampled from backyard poultry - poultry sector 4 from the risk areas in Republic of Macedonia i.e., areas where migratory waterfowl gathers) and 131 organs/cloacal swabs (for daily routine diagnostics) were tested by virological and molecular methods. The virological diagnosis was performed by isolation on embryonated chicken eggs, followed by macro hemagglutination, hemagglutination on microtiter plate and inhibition of hemagglutination. The molecular diagnosis was performed by RRT-PCR for M-gene. All samples were negative for AIV. However, 4 samples were positive on avian paramyxovirus 1 (Newcastle disease virus). The wild birds, as a reservoir for AIV were not included in the surveillance which on the other hand does not ensure complete information for the AIV circulation in R. Macedonia and should be further investigated. This fact imposes the need for overall national action to ensure the concept of early detection and response in case of emergence of avian influenza. [ABSTRACT FROM AUTHOR]