Labrini V. Athanasiou

A cross-sectional sero-epidemiological study of canine leishmaniasis in Greek mainland.

Canine leishmaniasis is endemic in Greece as in other countries of the Mediterranean basin. In this study, the regional prevalence of canine seropositivity to Leishmania spp. in Greek mainland was simultaneously assessed in 7 different regions. A total of 2620 serum samples were collected from clinically healthy dogs and were tested for anti-L. infantum antibodies by indirect immunofluorescence antibody test (IFAT) and enzyme linked immunosorbent assay (ELISA). A high degree of agreement (κ=0.96) was observed between these two tests. The seroprevalence rate in the whole study population was nearly 20%, being highest among dogs living in Attiki (30.12%) and lowest for those living in Florina (2.05%). There was no difference in terms of the gender of the dogs, their length of hair coat or their utility; on the contrary, seroprevalence rates were significantly higher among dogs in the age groups of 1-3 years (23.39%) and 3-9 years (23.35%) than in younger (2.26%) or older (6.03%) dogs. Epidemiological data on the seroprevalence of canine leishmaniasis provide indirect information on the prevalence of the infection and the disease and are necessary to implement and then to evaluate the effectiveness of control measures.

Serological and molecular investigation into the role of wild birds in the epidemiology of West Nile virus in Greece

BackgroundA West Nile virus (WNV) disease outbreak occurred in 2010 in northern Greece with a total of 262 laboratory-confirmed human cases and 35 deaths. A serological and molecular surveillance was conducted on samples of hunter-harvested wild birds prior to and during the outbreak.FindingsSerum and tissue samples from 295 resident and migratory wild birds, hunter-harvested during the 2009–2010 and 2010–2011 hunting seasons at the epicenter of the outbreak in northern Greece, were tested for the presence of WNV-specific antibodies by immunofluorescence assay and virus neutralization test. WNV neutralizing antibodies were detected in 53 avian samples. Fourteen positive sera were obtained from birds hunter-harvested up to 8 months prior to the human outbreak. Specific genetic determinants of virulence (His249Pro NS3 mutation, E-glycosylation motif) were recognized in a WNV lineage 2 strain isolated from a hunter-harvested Eurasian magpie and a nucleotide mismatch was revealed between this strain and a mosquito WNV strain isolated one month earlier in the same area.ConclusionsThis is the first report regarding exposure of wild birds to WNV prior to the 2010 outbreak, in Greece. Results provide evidence of the implication of wild birds in a local enzootic cycle that could allow maintenance and amplification of the virus before and during the outbreak. Findings of past exposure of migratory birds to WNV upon their arrival in Greece during autumn migration, suggest avian species with similar migration traits as candidates for the introduction of WNV into Greece. The possibility that an endemic circulation of WNV could have caused the outbreak, after an amplification cycle due to favorable conditions cannot be excluded.

The effect of a natural feed additive (Macleaya cordata), containing sanguinarine, on the performance and health status of weaning pigs

The aim of this study was to investigate the efficacy of Sangrovit(®) , a plant-derived feed additive, given throughout the nursery stage via feed at 15 ppm and 50 ppm, on the health status and performance of weaners against negative controls. In a 900-sow farrow-to-finish farm, a total of 864 piglets were divided into three groups: (i) negative controls (NC); (ii) Sang 15: same feed as NCs, plus 15 g Sangrovit(®) /t of feed; (iii) Sang 50: same feed as NCs plus 50 g Sangrovit(®) /t of feed. The results indicated that administration of 50 ppm(-) Sangrovit(®) had the most beneficial effects on growth performance in weaning pigs. Specifically there was increase of body weight and average daily gain, as well as reduction of feed conversion ratio. Blood analysis from the Sang groups and especially the Sang 50 group revealed low values of haptoglobin and serum amyloid A.

A serosurvey for selected pathogens in Greek European wild boar

Objectives Serum samples, collected from 94 European wild boar (Sus scrofa) during the hunting seasons 2006 -2010 from different regions of Greece, were examined in order to estimate the role of these wildlife species as reservoir of pathogens important for livestock and/or public health. Materials and Methods The assays used for this purpose were commercial indirect ELISA for the detection of antibodies against porcine circovirus type 2 (PCV-2), porcine reproductive and respiratory syndrome (virus) (PRRSV), Aujeszkys disease virus (ADV), influenza A (IA) virus, Actinobacillus pleuropneumoniae, Mycoplasma hyopneumoniae, Salmonella species, Trichinella species and indirect immunofluorescence antibody test for the detection of antibodies against Toxoplasma gondii and Neospora caninum. Results Antibodies against PCV-2, PRRSV, ADV, IA virus,A. pleuropneumoniae, M. hyopneumoniae,Salmonella species, Trichinella species, T. gondii and N. caninum were detected in 19.1 per cent, 12.8 per cent, 35.1 per cent, 1.1 per cent, 57.4 per cent, 0 per cent, 4.3 per cent, 6.4 per cent, 5.2 per cent and 1.1 per cent of the samples, respectively. Cluster analysis revealed a hot spot of seropositivity near Bulgarian border; seropositivity to ADV was more common among female animals. Conclusions These results indicate exposure of wild boar to most of the above-mentioned pathogens, raising concern about the possibility that these species may pose a significant health risk for livestock and/or humans.

Use of wild bird surveillance, human case data and GIS spatial analysis for predicting spatial distributions of West Nile virus in Greece.

West Nile Virus (WNV) is the causative agent of a vector-borne, zoonotic disease with a worldwide distribution. Recent expansion and introduction of WNV into new areas, including southern Europe, has been associated with severe disease in humans and equids, and has increased concerns regarding the need to prevent and control future WNV outbreaks. Since 2010, 524 confirmed human cases of the disease have been reported in Greece with greater than 10% mortality. Infected mosquitoes, wild birds, equids, and chickens have been detected and associated with human disease. The aim of our study was to establish a monitoring system with wild birds and reported human cases data using Geographical Information System (GIS). Potential distribution of WNV was modelled by combining wild bird serological surveillance data with environmental factors (e.g. elevation, slope, land use, vegetation density, temperature, precipitation indices, and population density). Local factors including areas of low altitude and proximity to water were important predictors of appearance of both human and wild bird cases (Odds Ratio = 1,001 95%CI = 0,723–1,386). Using GIS analysis, the identified risk factors were applied across Greece identifying the northern part of Greece (Macedonia, Thrace) western Greece and a number of Greek islands as being at highest risk of future outbreaks. The results of the analysis were evaluated and confirmed using the 161 reported human cases of the 2012 outbreak predicting correctly (Odds = 130/31 = 4,194 95%CI = 2,841–6,189) and more areas were identified for potential dispersion in the following years. Our approach verified that WNV risk can be modelled in a fast cost-effective way indicating high risk areas where prevention measures should be implemented in order to reduce the disease incidence.

Evaluation of indirect immunofluorescence antibody test and enzyme-linked immunosorbent assay for the diagnosis of infection by Leishmania infantum in clinically normal and sick cats.

Abstract Cats that live in areas where canine and human leishmaniosis due to Leishmania infantum is endemic may become infected and may develop anti-Leishmania antibodies. In this study 50 clinically normal and 50 cats with cutaneous and/or systemic signs that lived in an endemic area and had been previously examined for infection by L. infantum using PCR in four different tissues were serologically tested for the presence of anti-Leishmania IgG (IFAT and ELISA) and IgM (IFAT). The aim was to compare the results of IFAT, ELISA and PCR and to investigate the possible associations between seropositivity to Leishmania spp and signalment, living conditions, season of sampling, health status of the cats, and seropositivity to other infectious agents. Low concentrations of anti-Leishmania IgG were detected by IFAT in 10% of the cats and by ELISA in 1%, whereas anti-Leishmania IgM were detected by IFAT in 1%. There was disagreement between the results of IFAT and ELISA for anti-Leishmania IgG (P = 0.039) and between all serological tests and PCR (P < 0.001). The diagnostic sensitivity of all serological tests, using PCR as the gold standard, was very low, but ELISA and IFAT for anti-Leishmania IgM had 100% specificity. The diagnostic sensitivity of all serological tests could not be improved by changing the cut-off values. Seropositivity for Leishmania spp was not associated with signalment, living conditions, season of sampling and health status of the cats or with seropositivity to feline leukemia virus, feline immunodeficiency virus, feline coronavirus, Toxoplasma gondii and Bartonella henselae. In conclusion, because of their low sensitivity and very high specificity two of the evaluated serological tests (ELISA for anti-Leishmania IgG and IFAT for anti-Leishmania IgM) may be useless as population screening tests but valuable for diagnosing feline infection by L. infantum.

Treatment of canine leishmaniosis with aminosidine at an optimized dosage regimen: a pilot open clinical trial.

Leishmaniosis due to Leishmania infantum (Syn: L. chagasi) is one of the most common diseases of dogs in Mediterranean countries and also has zoonotic potential. The aim of this study was to evaluate the efficacy of an optimized dosage regimen of aminosidine for the treatment of canine leishmaniosis (CanL) in terms of clinical remission, restoration of clinicopathological abnormalities, evolution of antibody titer, lymph node and bone marrow parasitic density and of PCR-based parasitological cure. Twelve non-uremic dogs without proteinuria, presenting clinical signs of CanL were included in the study. The diagnosis was confirmed by serology, microscopy and PCR of lymph node and bone marrow samples. Aminosidine was administered subcutaneously at the dose of 15 mg/kg body weight, once daily, for 21 consecutive days. A partial remission of the clinical signs, amelioration of clinicopathological abnormalities such as anemia, lymphopenia, hyperproteinemia, hyperglobulinemia, and reduced albumin/globulin ratio and reduced lymph node and bone marrow parasitic density were witnessed, although parasitological cure was not achieved. Since data are not supportive enough for the use of aminosidine as an alternative treatment, a large-scale controlled clinical trial using this optimized dosage regimen of aminosidine is warranted to compare efficacy against currently used drugs.

Bluetongue in small ruminants: An opinionated review, with a brief appraisal of the 2014 outbreak of the disease in Greece and the south-east Europe.

Bluetongue is an arthropod-borne viral disease of ruminants, especially of sheep, caused by Bluetongue virus, which belongs to the genus Orbivirus of the family Reoviridae and is classified into 26 antigenically distinct serotypes. Once thought to be restricted in Africa and parts of the Middle East, bluetongue has now become a concern in sheep-rearing countries around the world. In the past 10 years, severe outbreaks have occurred in Europe with important economic consequences; of these, the 2006-20008 outbreak in Europe was caused by a serotype 8 strain and the 2014 outbreak in Greece and the other countries of south-east Europe was caused by a serotype 4 strain, suggested to be a reassortant strain with genome segments from lineages of serotype 1, 2 and 4. Immunisation campaigns can be implemented for successful control and limiting of the disease. Nevertheless, in both of the above outbreaks, late application of vaccinations led to a wide spread of the disease, which subsequently resulted in significant losses in livestock in the affected regions. In view of that, standardisation of control measures in the future will be beneficial for efficiently limiting outbreaks of the disease.

Emerging Trichinella britovi infections in free ranging pigs of Greece

Trichinella infections in humans and pigs have been documented in Greece since 1945 and a high prevalence of infection in pigs occurred in the 1950s. Up to 1984 only sporadic infections in humans were documented, and this zoonosis was not considered as a public health problem until 2009 when a human outbreak caused by the consumption of pork from an organic pig farm occurred. In the present study, we describe the re-emergence of Trichinella spp. infections in free-ranging pigs from organic farms of 3 counties (Dramas, Evros and Kavala) in Northern-Eastern Greece during the period 2009-2012. Totally 37 out of 12,717 (0.29%) free-ranging pigs which were tested during the period in question, were positive for Trichinella spp. larvae. The etiological agent was identified as Trichinella britovi. The average larval burden was 13.7 in the masseter, 6.2 in the foreleg muscles and 7.5 in the diaphragm. The 37 positive animals originated from seven free range pig farms. The practice of organic pig production systems in Greece has grown in popularity over the last years due to the increasing interest of consumers for products considered as traditional. However, this type of pig production increases the risk for Trichinella spp. infections, since animals can acquire the infection by feeding on carcasses or the offal of hunted or dead wild animals. The awareness and education of hunters and farmers is extremely important to reduce the transmission among free ranging pigs and the risk for humans.

Cross-Sectional Serosurvey and Risk Factors Associated with the Presence of Toxoplasma gondii Antibodies in Pigs in Greece

INTRODUCTION Toxoplasmosis is a worldwide reported zoonotic infection caused by the protozoon Toxoplasma gondii. Pigs may become infected by ingesting feed or water contaminated with cat faeces, by cannibalism, and/or by eating infected rodents. T. gondii infected edible tissues of pigs are a source of infection for humans. This study was undertaken to detect seropositivity of pigs against T. gondii and identify possible risk factors as well as to compare two serological methods. METHODS A total of 609 blood samples were collected from 10% of the sows from 65 pig farms located in mainland Greece. Information about the geographical location, size, and biosecurity level of the herd was recorded. Samples were tested for the presence of antibodies against T. gondii employing the indirect fluorescent antibody test and the enzyme-linked immunosorbent assay. RESULTS Twenty-six positive samples were detected (4.3%) in 17 out of the 65 farms tested (26.2%). The average seroprevalence in affected herds was 26.8% (95% confidence interval 21.0-32.6%). Among the categorical data evaluated, the biosecurity level (odds ratio 0.17, p < 0.01) and the geographical characteristics (odds ratio 13.55, p < 0.05) significantly affected the presence of toxoplasmosis in the herd. Toxoplasmosis was recorded in significantly higher percentages in mountainous farms as compared to lowlands (p < 0.05) and in those with poor biosecurity levels than in those with very good (p < 0.05). A very good agreement (κ = 0.958) was observed between the two serological methods. CONCLUSION The presence of antibodies in pigs is indirect information on the risk of the infection and an indication of the necessity of biosecurity measures to be taken in order to control infection at least in the areas at highest risk.