Emergence of Lumpy Skin Disease in Greece, 2015.

Lumpy skin disease was first reported in the EU, Greece, in August 2015. Until the end of December 2015, six regional units have been affected in northern Greece and the island of Limnos. This article presents the epidemiological situation, the diagnosis, the control measures including emergency vaccination and the preliminary conclusions from the experience gained.

Re-emergence of animal rabies in northern Greece and subsequent human exposure, October 2012 - March 2013.

Greece has been rabies-free since 1987 with no human cases since 1970. During 2012 to 2013, rabies has re-emerged in wild and domestic animals in northern Greece. By end March 2013, rabies was diagnosed in 17 animals including 14 red foxes, two shepherd dogs and one cat; 104 subsequent human exposures required post-exposure prophylaxis according to the World Health Organization criteria. Human exposures occurred within 50 km radius of a confirmed rabies case in a wild or domestic animal, and most frequently stray dogs were involved.

Epizootiological investigation of the most important infectious equine diseases in Greece.

During the period 2001 to 2008, a total of 7,872 equine sera were tested at the Centre of Veterinary Institutes of Athens. Antibodies against seven infectious diseases of equids were determined: equine infectious anaemia (EIA), African horse sickness (AHS), equine viral arteritis (EVA), West Nile encephalitis (WNE), glanders, piroplasmosis and dourine. Tests for the four viral diseases found 4.5% seropositivity for EIA, 0% for AHS, 3.3% for EVA and 4% for WNE. All sera tested for glanders antibodies were negative. Tests for piroplasmosis detected antibodies against T. equi and B. caballi in 12.9% and 1.3% of the sera, respectively. No sample tested positive for dourine. The results of this epidemiological survey provide strong evidence that Greece is free from the diseases of AHS, glanders and dourine.

The diversity of juvenile sarcoidosis symptoms.

We report a case of juvenile sarcoidosis, emphasizing the variety of clinical manifestations. The child had uveitis, which is among the most common manifestations of the disease. However, fever of unknown origin, glomerulonephritis and lymphadenopathy were also noticed, underscoring the diversity of the clinical spectrum of the disease.

Overview of bluetongue in Greece.

The history and epizootiology of bluetongue (BT) in Greece are described in detail. Three major epidemics of BT occurred in Greece, the first in 1979, due to bluetongue virus (BTV) serotype 4, in 1998-1999 due mainly to BTV-4 and BTV-9 and less to BTV-16 and in 2001 due mainly to BTV-1. The evolution of the disease, surveillance and control measures are presented, as well as the distribution of the BTV serotypes isolated.

S10 segment sequence analysis of some Greek bluetongue virus strains.

Sequence analyses of the non-structural protein gene NS3/NS3A of eight Greek bluetongue (BT) virus (BTV) field isolates from the 1979 and 1999-2001 epizootics provide preliminary molecular data on the epidemiology of BT in Greece. These isolates from infected sheep belonged to serotypes BTV-1, BTV-4, BTV-9 and BTV-16. Phylogenetic analysis of the NS3/NS3A gene segregated these Greek isolates of BTV into two monophyletic groups. The first group was formed by all isolates of BTV-4; all were identical in their sequences, regardless of the area and year of isolation in Greece, and clustered with strains from Tunisia and Corsica. The isolates of BTV-1, BTV-9 and BTV-16 segregated into a second monophyletic group and clustered with Asian strains, showing a high homology (97-99%). From an epidemiological point of view, these preliminary results infer that one group of isolates is Mediterranean, whilst the second appears to be of Asian origin.

VP2 gene sequence analysis of some isolates of bluetongue virus recovered in the Mediterranean Basin during the 1998-2002 outbreak.

Since 1998, five serotypes of bluetongue virus (BTV), BTV-1, BTV-2, BTV-4, BTV-9 and BTV-16, have been reported in countries surrounding the Mediterranean Basin. Preliminary data on the sequencing analysis of the VP2-genes of BTV isolates recovered during the 1998-2002 epizootic of BT in Italy, Greece and Israel were studied. The VP2-genes of the Italian BTV-2 and BTV-9, Greek BTV-4 and BTV-9, Israeli BTV-4 and BTV-16 and South African BTV-2, BTV-4, BTV-9 and BTV-16, together with those of their corresponding South African serotype reference and vaccine strains, were cloned and the sequences of their terminal ends determined. These sequences, as well as those of all BTV VP2-gene sequences currently available on GenBank, were used to compile a phylogenetic tree to determine the probable geographic origins of the BTV incursions into Europe. The Italian isolates included in this study were from different regions, animal hosts and years (2000-2002). The results demonstrated that sequencing of the terminal end of the VP2-gene of BTV can be used for topotyping. According to the phylogenetic analysis, the Italian BTV-2 and BTV-9 isolates were stable across all species, irrespective of geographic origin and year of isolation. The sequencing data of the Italian isolates were identical to those of a BTV-2 isolate from Corsica. There was 97% homology between the Italian and Corsican BTV-2 isolates and the BTV-2 vaccine and reference isolates from South Africa. Italian BTV-9 isolates were also identical to the Greek BTV-9 isolates (99% homology). Surprisingly these BTV-9 isolates had only 67% homology with the reference BTV-9 isolate from South Africa. Conversely, BTV-9 field isolates from Australia and elsewhere in Europe had 89% homology with the Italian isolate at the nucleic acid level. Greek and Israeli BTV-4 isolates were almost identical (98% homology) and shared a 90% homology with the BTV-4 South African reference and vaccine strains. Israeli BTV-16 and South African BTV-16 reference strains were also similar. From these results, it may be concluded that Italian and Corsican BTV-2, Israeli and Greek BTV-4, and South African and Israeli BTV-16 had a common origin. The Greek BTV-9 isolate had more than 99% homology with the isolates from Italy, indicating these isolates to have had a common origin. The European BTV-9 isolates, grouped as 'eastern isolates', were more similar to the Australian isolates than to the South African reference strains.

Bluetongue laboratory diagnosis: a ring test to evaluate serological results using a competitive ELISA kit.

The occurrence of bluetongue (BT) in Italy prompted an increase in disease surveillance. Thus a competitive enzyme-linked immunosorbent assay (c-ELISA) to detect immunoglobulins to BT virus (BTV) was developed and distributed amongst 27 laboratories comprising the Italian veterinary diagnostic laboratories network to screen field sera. This ring test enabled comparison of the results and the evaluation of the reproducibility of the method. The c-ELISA developed by the National Reference Centre for Exotic Diseases (c-ELISA-IZSA&M) was compared also against a commercially available c-ELISA. In addition, results obtained by the Centre of Athens Veterinary Institutions are presented.

The Greek version of the Childhood Health Assessment Questionnaire (CHAQ) and the Child Health Questionnaire (CHQ).

We report herein the results of the cross-cultural adaptation and validation into the Greek language of the parent's version of 2 health related quality of life instruments. The Childhood Health Assessment Questionnaire (CHAQ) is a disease specific health instrument that measures functional ability in daily living activities in children with juvenile idiopathic arthritis (JIA). The Child Health Questionnaire (CHQ) is a generic health instrument designed to capture the physical and psychosocial well-being of children independently from the underlying disease. The Greek CHAQ CHQ were fully validated with 3 forward and 3 backward translations. A total of 143 subjects were enrolled: 82 patients with JIA (28% systemic onset, 24% polyarticular onset, 10% extended oligoarticular subtype, and 38% persistent oligoarticular subtype) and 61 healthy children. The CHAQ clinically discriminated between healthy subjects and JIA patients, with the systemic, polyarticular and extended oligoarticular subtypes having a higher degree of disability, pain, and a lower overall well-being when compared to their healthy peers. Also the CHQ clinically discriminated between healthy subjects and JIA patients, with the systemic onset, polyarticular onset and extended oligoarticular subtypes having a lower physical and psychosocial well-being when compared to their healthy peers. In conclusion the Greek version of the CHAQ-CHQ is a reliable, and valid tool for the functional, physical and psychosocial assessment of children with JIA.

Sheep poxvirus identification from clinical specimens by PCR, cell culture, immunofluorescence and agar gel immunoprecipitation assay.

Some 40 clinical specimens of skin lesions from sheep pox suspected cases were investigated by four different diagnostic assays: PCR, virus isolation in lamb testis cell cultures, direct immunofluorescent assay (DIFA) and antigen detecting agar gel immune precipitation test (AGIPT). All the specimens were positive by PCR and virus isolation, 29 were positive by DIFA and 16 by AGIPT. Using virus isolation on cell cultures as the gold standard, the PCR sensitivity was 100%, while that of DIFA and AGIPT was 73% and 40%, respectively. Skin samples with orf lesions or normal skin biopsies were PCR-negative. Cross-reactions with orf virus were observed in three samples only in the AGIPT assay. The PCR described combines high specificity and sensitivity with speed. PCR was therefore shown to be the method of choice for sheep poxvirus diagnosis directly from clinical specimens.

Benign hypermobility syndrome in Greek schoolchildren.

Since the incidence of benign hypermobility syndrome is significantly high in otherwise healthy children, paediatricians should consider this benign entity when they evaluate musculoskeletal complaints in childhood.

Detection of sheep poxvirus in skin biopsy samples by a multiplex polymerase chain reaction.

The development of a multiplex polymerase chain reaction (PCR) method with amplification of capripoxvirus in a single-step procedure from skin biopsies using three primer pairs, two specific for capripoxvirus and one specific for alpha-tubulin is described. A sensitive multiplex PCR was achieved by optimization of parameters such as the primer concentrations, magnesium and dNTPs concentrations. False negative results that sometimes arise due to inhibitors of DNA amplification may be avoided by the inclusion in the assay of alpha-tubulin primers. The results reported on 42 skin biopsies from sheep suspected to have poxvirus infection, indicated that the assay could monitor simultaneously DNA extraction from skin biopsy samples and allow improved detection of capripoxvirus within 24 h of specimen receipt in the laboratory.

Sheep poxvirus identification by PCR in cell cultures.

A simple, rapid and specific diagnostic polymerase chain reaction (PCR) method was developed for sheep poxvirus identification. The primers used were from the sequenced genomes of the capripox viruses KS-1 and InS-1. Six different sheep pox isolates were tested against two orf (parapox) and three animal herpesviruses as controls. Material from uninfected cell cultures was also used as control. The sensitivity of the PCR was approximately equivalent with each of the two primers and for the six sheep pox isolates. All the negative control virus DNAs were negative and differed clearly from those of the sheep pox strains.