First report of fatal autochthonous angiostrongylosis in a dog in Greece.

Canine angiostrongylosis caused by Angiostrongylus vasorum is a life-threatening disease which is emerging in regions of Europe. Thus, there is the merit for a continuous epidemiological surveillance in dog populations. This is the first description of a clinical autochthonous case of canine angiostrongylosis in Greece. A 7-month-old, male, mixed-breed dog was presented with progressively worsening anorexia, respiratory distress, coughing, bleeding diathesis and succumbed four days post admission. Gross post mortem examination revealed numerous nodular fistulated lesions in the lungs, and pulmonary cytology and histopathology showed a verminous pyogranulomatous pneumonia. The definitive diagnosis was based on the morphological identification of first stage larvae (L1) retrieved in the feces by the Baermann method, the detection of the circulating antigen by an in-clinic test and the molecular identification of L1. This report underlines the epidemiological and clinical implications, as well as the infection risks when the index of clinical suspicion is low and the disease is unexpected in a country.

Human trichinellosis caused by Trichinella britovi in Greece, and literature review.

Trichinellosis is a cosmopolitan zoonotic parasitic disease caused by the nematodes of the genus Trichinella, through the consumption of raw or semi-raw infected meat from swine, horses and wild animals. This disease has been sporadically reported in Greece since 1946. The aim of the present study was to describe a trichinellosis case in a patient hospitalized in northern Greece, in 2017. A 47-year-old male was admitted to hospital with intense generalized myalgia, periorbital swelling, fever, exhaustion and anorexia. Biochemical and haematological profile showed eosinophilia and elevated creatine phosphokinase (CPK). Anti-Trichinella spp. IgG and IgM antibodies were detected by serology and Trichinella spp. larvae were found in two muscle biopsies by compressorium and histological examination. A larva collected from the muscle biopsy was identified as Trichinella britovi by polymerase chain reaction (PCR). Albendazole (400 mg twice per day × 10 days) was administered and the clinical condition of the patient promptly improved. This is the first identification of T. britovi in a patient in Greece.

The role of an attenuated anticoccidial vaccine on the intestinal ecosystem and on the pathogenesis of experimental necrotic enteritis in broiler chickens.

The objective of the present study was to investigate the effect of an attenuated anticoccidial vaccination on the intestinal ecosystem and on the pathogenesis of experimental necrotic enteritis (NE) in broiler chickens. Two hundred and forty 1-day-old Cobb 500 broiler chickens were randomly allocated to four treatment groups according to the following experimental design: control Group N; Group PN, where birds were vaccinated with anticoccidial vaccine; Group M, where birds were challenged with Clostridium perfringens and with Eimeria maxima; and Group PM, where birds were both vaccinated and challenged. From each bird, the intestine, gizzard and liver were scored for gross NE lesions. Intestinal digesta were collected for pH and viscosity determination. Samples from the gastrointestinal tract and liver were taken for microbiological analysis. Evaluation of the experimental data revealed that Group M had significantly higher overall mean NE intestinal lesions compared with Group PM. Viscosity values of jejunum digesta as well as pH values of the duodenum, jejunum and ileum digesta in Group M were significantly lower compared with Group PM. C. perfringens counts in the caeca of Group PM were significantly lower compared with Group M. The milder decrease of pH and viscosity values of intestinal content and the reduction of C. perfringens counts in the caeca in challenged and vaccinated birds may explain the lower score of NE gross intestinal lesions and may suggest a positive effect on intestinal ecosystem and a significant protective effect of attenuated anticoccidial vaccination against NE in a subclinical experimental model.

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.

Vaccination with Trichinella spirallis antigens increases CD8+ peripheral T cells and enhances the Th2 immune response in Leishmania infantum challenged mice.

In this study we investigate the effect of Trichinella spiralis vaccination on immune responses elicited in BALB/c mice challenged subcutaneously with 0.5 x 10 6 of Leishmania infantum promastigotes. Secretion of specific anti-L. infantum antibodies and changes in the number of CD4+, CD8+ T cell and CD19+ B cells in the peripheral blood were tested for the evaluation of immune responses. Immunization with low amounts of T. spiralis antigens induced depression in anti-Leishmania specific antibodies of the IgG1 isotype, while no changes in the number of CD4+ and CD8+ T cell subpopulations or CD19+ B cells were observed. In contrast, high amounts of T. spiralis antigens induced an enhancement in anti-Leishmania specific antibodies of total IgG and IgG1 isotype, increase of CD8+ T cell number and activation of CD19+ B cells, indicated by the co-expression of CD69 marker. Our results suggest that immunization with a certain dose of T. spiralis antigens in experimentally challenged mice with L. infantum leads to an increase of peripheral CD8+ T cells which are responsible for the control of L. infantum infection, although a simultaneous enhancement in Th2-type of immune response is also observed.

Long-term evaluation of patients with hydatidosis treated with albendazole and praziquantel.

Hydatidosis is a usually asymptomatic chronic disease. In most patients who undergo surgery, hydatidosis is not resolved due to high recurrence rate. However, long-term treatment with albendazole has been found to have a significant efficacy that has been further improved when albendazole is combined with praziquantel and fat-rich diet. In this study a retrospective evaluation of the outcome of hydatidosis in 70 patients, was performed. In group A, a combined chemotherapy of albendazole plus praziquantel was given after surgical removal of cysts. In group B chemotherapy alone was administered without surgery. Sera of all patients were assayed for IgG, IgM, IgA and IgE antibodies by ELISA. In addition, ultrasonography (US) and/or computerized tomography (CT) scans were performed every 3 months for 18 months, and then, each year until the end of follow-up. The difference between the two kinds of treatment used in the present study was found to be not significant, nor was the difference of the shrinkage and extended calcification of the HCs between the two groups. However, the difference of the shrinkage of the HCs of more than 80%, as well as the extended calcifications of the cysts between the two groups were found to be statistically significant. In all patients high levels of IgG and IgA were detected, while IgE in group A and/or IgM in group B were marginally detected above the background level throughout the study. Level of IgG was strongly fluctuated and significantly decreased at 11.7 years after the end of chemotherapy, or at 8.5 years after relapses in group A, while was dramatically decreased at 3.6 years after the termination of chemotherapy in group B. Relapses occurred in 11.4% of patients within the first six months after end of chemotherapy. After additional chemotherapy with albendazole for 3-6 months, all of them were considered cured at 8.5 years of follow up.

Comparison of infection rates of Oestrus ovis between sheep and goats kept in mixed flocks.

Oestrosis is a nasal myiasis of sheep and goats caused by larvae of the fly Oestrus ovis and can lead to severe clinical signs, which together with the disturbance caused by the adult fly may result into serious economic losses. Infection rates and larval burdens are always higher in sheep than in goats after either natural or artificial infestation. The aim of this study was to compare the host preference of the adult fly O. ovis between sheep and goats in mixed flocks, where they are kept together under the same husbandry conditions and hence, are very similarly exposed to the fly preference. Blood sera samples were collected from a total of 397 sheep and 335 goats, from 43 mixed flocks located at different regions of Greece. Antibodies specific to O. ovis IgG were measured by ELISA. A flock was considered positive when at least one individual was positive, i.e. showed a seropositivity of >or=20% in relation to positive control sera. A total of 193 (48.6%) sheep and 58 (17.9%) goats were found to be seropositive against O. ovis. Thirty-eight (88.4%) out of 43 flocks had at least one seropositive animal. The mean seroconversion against O. ovis in animals from the different flocks was 38.6% and 13.6% for sheep and goats, respectively, whereas the variance of infection within each flock was 0-100%. The mean seropositivity between sheep that were found to be positive or negative was 60.6% and 5.4%, respectively, whereas the corresponding values between goats were 35.2% and 5.2%, respectively. No significant difference in the seroconversion values was noted between flocks from the different areas (P=0.817), whereas a very significant difference was observed between animal species (P=0.001). However, there was no significant difference when seroconversion comparisons were made within samples of the same animals species, sheep or goats from different flocks of all the regions included in the study (P=0.695). The results of this study clearly demonstrate that O. ovis has a widespread distribution in Greece, and the seroprevalence is significantly higher in sheep than goats (P=0.001).

CT and MR imaging findings in cerebral toxocaral disease.

We report serial MR findings in a 54-year-old woman with eosinophilic meningoencephalitis due to Toxocara canis infection, a parasitic disease contracted through exposure with soil contaminated by the eggs of the roundworm. MR imaging revealed several enhancing subcortical and white matter lesions in both lobes. Antihelminthic chemotherapy yielded marked improvement of the neurologic deficits and cerebral lesions. The specific MR findings-low signal intensity on T1-weighted images, high signal intensity on T2-weighted images, and contrast enhancement-and the clinical and epidemiologic features of CNS involvement are herein reviewed.

Optimization of random-amplified polymorphic DNA producing amplicons up to 8500 bp and revealing intraspecies polymorphism in Leishmania infantum isolates.

In the case of investigation of polymorphism in closely related strains, the highest possible complexity of the patterns obtained by random-amplified polymorphic DNA PCR (RAPD-PCR) is required to assure revealing of limited polymorphism. In the present work, most parameters (reaction components concentration, additives, different polymerases, and thermal profiles) affecting RAPD-PCR were examined, in an effort to increase pattern complexity. A long PCR thermal profile, betaine as cosolvent, and Dynazyme EXT polymerase produced longer amplicons and higher pattern complexity, revealing polymorphism among Leishmania infantum isolates from infected dogs originating from Northern Greece (Macedonia).