Validation of an improved reference freeze-dried direct agglutination test for detecting leishmaniasis in the canine reservoir.

Proper identification and management of post-kala-azar dermal leishmaniasis (PKDL) and canine leishmaniasis (CanL) cases are among the prerequisites to the effective control of visceral leishmaniasis worldwide. Unlike PKDL, CanL still awaits effective improvement because of its cryptic nature, absence of parasites in lesions or lymph nodes and not complete sensitivity of some diagnostic tools in use. Because of the need for certain skills and equipment, both the liquid direct agglutination test and freeze-dried direct agglutination test (FD-DAT) versions are, in comparison with the indirect immunofluorescence antibody test (IFAT) or enzyme-linked immunosorbent assay (ELISA), practical and feasible diagnostic alternatives.

Data Independent Acquisition Reveals In-Depth Serum Proteome Changes in Canine Leishmaniosis.

Comprehensive profiling of serum proteome provides valuable clues of health status and pathophysiological processes, making it the main strategy in biomarker discovery. However, the high dynamic range significantly decreases the number of detectable proteins, obstructing the insights into the underlying biological processes. To circumvent various serum enrichment methods, obtain high-quality proteome wide information using the next-generation proteomic, and study host response in canine leishmaniosis, we applied data-independent acquisition mass spectrometry (DIA-MS) for deep proteomic profiling of clinical samples.

Severe Pulmonary Fascioloidosis in a Wild Mouflon () - a Case Report.

A wild male mouflon () was shot due to the observed weakness. Necropsy revealed consolidated lungs and traces of black pigment and fibrin on the liver. On the cut surface, a juvenile fl uke was found in the lungs, while traces of destroyed fl ukes' migratory channels were found in the liver.

Field surveys in Croatia and North Macedonia reveal two novel phleboviruses circulating in sandflies.

Sandfly-borne phleboviruses are distributed widely throughout the Mediterranean Basin, presenting a threat to public health in areas where they circulate. However, the true diversity and distribution of pathogenic and apathogenic sandfly-borne phleboviruses remains a key issue to be studied. In the Balkans, most published data rely on serology-based studies although virus isolation has occasionally been reported.

First record of chronic infection in roe deer ().

Fascioloidosis is an allochthonous parasitic disease in Europe caused by the digenean trematode . The final hosts of in Europe are defined as definitive, aberrant and dead-end. Roe deer are aberrant hosts in which juvenile flukes permanently migrate through the liver parenchyma.

Sand flies (Diptera: Psychodidae) in eight Balkan countries: historical review and region-wide entomological survey.

Background: Sand flies (Diptera: Psychodidae) are medically important vectors of human and veterinary disease-causing agents. Among these, the genus Leishmania (Kinetoplastida: Trypanosomatidae), and phleboviruses are of utmost importance. Despite such significance, updated information about sand fly fauna is missing for Balkan countries where both sand flies and autochtonous leishmaniases are historically present and recently re-emerging.

The First Description of (Issaitshikoff & Rajewskaya, 1928) in Roe Deer from Croatia.

Genus , Viborg 1795, comprises 46 species that parasitize in the peritoneal cavity of Artiodactyla, Perissodactyla and Hyracoidea. The majority of these infections pass unnoticed, but occasionally they can induce severe peritonitis or neurological signs in aberrant hosts and, rarely, even in humans. In this paper we describe for the first time the finding of in roe deer in Croatia.

Seroprevalence of vector-borne pathogens in dogs from Croatia.

Canine vector-borne pathogens (VBPs) are a group of globally distributed and rapidly spreading microorganisms transmitted by arthropods. In the present survey, we investigated Anaplasma spp., Ehrlichia canis, Dirofilaria immitis and Borrelia burgdorferi sensu lato seroprevalence between three groups of dogs (asymptomatic, suspected and deceased) from continental and coastal regions of Croatia with the commercial point-of-care SNAP®4Dx®Plus for VBPs.

First identification of Echinococcus multilocularis in golden jackals in Croatia.

Alveolar echinococcosis, caused by the tapeworm Echinococcus multilocularis, is one of the world's most dangerous zoonosis and an emerging disease with growing incidence in humans. The disease has been reported in new areas and host species in the last two decades, and the primary hosts of the parasite - red fox, golden jackal and grey wolf - are expanding their distribution in Europe. Here we report the morphological and molecular identification of Echinococcus multilocularis tapeworms in one of 29 carcasses of adult golden jackals in Croatia, where the only previous report of the parasite was in red foxes in 2016.

Surface Proteome Biotinylation Combined with Bioinformatic Tools as a Strategy for Predicting Pathogen Interacting Proteins.

Constant advancements in methodology and mass spectrometry instrumentation, genome sequencing and bioinformatic tools have enabled the identification of numerous pathogen proteomes. Identifying the pathogen interacting proteins by means of high-throughput techniques is key for understanding pathogen invasion and survival mechanisms and in such a way proposing specific proteins as pharmaceutical targets. Herein we describe the methodology for the enrichment and identification of pathogen surface proteome using cell surface protein biotinylation followed by LC-MS/MS and bioinformatic analyses of such data.

Direct evidence for an expanded circulation area of the recently identified Balkan virus (Sandfly fever Naples virus species) in several countries of the Balkan archipelago.

Background: Recently, Balkan virus (BALKV, family Phenuiviridae, genus Phlebovirus) was discovered in sand flies collected in Albania and genetically characterised as a member of the Sandfly fever Naples species complex. To gain knowledge concerning the geographical area where exposure to BALKV exists, entomological surveys were conducted in 2014 and 2015, in Croatia, Bosnia and Herzegovina (BH), Kosovo, Republic of Macedonia and Serbia.

Results: A total of 2830 sand flies were trapped during 2014 and 2015 campaigns, and organised as 263 pools.

Molecular identification of the rumen flukes Paramphistomum leydeni and Paramphistomum cervi in a concurrent infection of the red deer Cervus elaphus.

Paramphistomosis, caused by paramphistomid flukes, is a gastrointestinal parasitic disease of domestic and wild ruminants. Originally thought to be limited to the tropics and subtropics, the disease has recently been reported in temperate regions. Here we describe the concurrent infection of a red deer doe (Cervus elaphus) with Paramphistomum leydeni and Paramphistomum cervi.

When things go wrong: Cysticercus longicollis in an adult wild red fox (Vulpes vulpes).

First case of Cysticercus longicollis, larval stage of Taenia crassiceps, was diagnosed in a wild adult male red fox (Vulpes vulpes). The fox was killed by dogs at Nature Park Medvednica and presented to the University of Zagreb Faculty of Veterinary Medicine with history of being unable to run away and having skin lesions on legs that resembled to those of mange. Necropsy revealed whitish fluctuant mass full of cysticercus-like structures, surrounded by fibrous capsule and placed between the leg muscles, and numerous of spherical cysts in the subcutis and in the peritoneal cavity.

Thelazia callipaeda--first human case of thelaziosis in Croatia.

A case of the first ocular thelaziosis in a human male patient in Croatia is presented. Thelaziosis is a zoonosis caused by the nematodes of the genus Thelazia. In all, two types of the genus have been described as causes of infection in humans, Thelazia callipaeda and Thelazia californiensis.

Heterogeneity of Leishmania donovani parasites complicates diagnosis of visceral leishmaniasis: comparison of different serological tests in three endemic regions.

Diagnostic tests for visceral leishmaniasis that are based on antigens of a single Leishmania strain can have low diagnostic performance in regions where heterologous parasites predominate. The aim of this study was to investigate and compare the performance of five serological tests, based on different Leishmania antigens, in three endemic countries for visceral leishmaniasis. A total number of 231 sera of symptomatic and asymptomatic cases and controls from three endemic regions of visceral leishmaniasis in East Sudan, North India and South France were evaluated by following serological tests: rKLO8- and rK39 ELISA, DAT (ITMA-DAT) and two rapid tests of rK39 (IT LEISH) and rKE16 (Signal-KA).

[THE FIRST CASE OF OCULAR THELAZIASIS IN CROATIA].

The first case of ocular thelaziasis in a human male patient in Croatia is presented. Thelaziasis is a zoonosis caused by nematodes of the genus Thelazia, parasites of the conjunctival bags or tear ducts of mammals and birds. Two types of the genus Thelazia (T.

Genetic diversity and structure in Leishmania infantum populations from southeastern Europe revealed by microsatellite analysis.

Background: The dynamic re-emergence of visceral leishmaniasis (VL) in south Europe and the northward shift to Leishmania-free European countries are well-documented. However, the epidemiology of VL due to Leishmania infantum in southeastern (SE) Europe and the Balkans is inadequately examined. Herein, we aim to re-evaluate and compare the population structure of L.

Trypanosoma (Megatrypanum) melophagium in the sheep ked Melophagus ovinus from organic farms in Croatia: phylogenetic inferences support restriction to sheep and sheep keds and close relationship with trypanosomes from other ruminant species.

Trypanosoma (Megatrypanum) melophagium is a parasite of sheep transmitted by sheep keds, the sheep-restricted ectoparasite Melophagus ovinus (Diptera: Hippoboscidae). Sheep keds were 100% prevalent in sheep from five organic farms in Croatia, Southeastern Europe, whereas trypanosomes morphologically compatible with T. melophagium were 86% prevalent in the guts of the sheep keds.

Multilocus phylogeographical analysis of Trypanosoma (Megatrypanum) genotypes from sympatric cattle and water buffalo populations supports evolutionary host constraint and close phylogenetic relationships with genotypes found in other ruminants.

Species of the subgenus Trypanosoma (Megatrypanum) have been reported in cattle and other domestic and wild ruminants worldwide. A previous study in Brazil found at least four genotypes infecting cattle (Bos taurus), but only one in water buffalo (Bubalus bubalis). However, the small number of isolates examined from buffalo, all inhabiting nearby areas, has precluded evaluation of their diversity, host associations and geographical structure.

Liver enzymes and blood metabolites in a population of free-ranging red deer (Cervus elaphus) naturally infected with Fascioloides magna.

We investigated the effects of Fascioloides magna infection on the serum biochemistry values of the naturally infected red deer population in eastern Croatia. The investigation was performed on 47 red deer with F. magna infection confirmed patho-anatomically in 27 animals (57.

Cysteine proteases of Trypanosoma (Megatrypanum) theileri: cathepsin L-like gene sequences as targets for phylogenetic analysis, genotyping diagnosis.

Although Trypanosomatheileri and allied trypanosomes are the most widespread trypanosomes in bovids little is known about proteolytic enzymes in these species. We have characterized genes encoding for cathepsin L-like (CATL) cysteine proteases from isolates of cattle, water buffalo and deer that largely diverged from homologues of other trypanosome species. Analysis of 78 CATL catalytic domain sequences from 22 T.

A case of visceral leishmaniosis in a gray wolf (Canis lupus) from Croatia.

The southern habitats of Croatia's gray wolf (Canis lupus) population are found in central and southern parts of Dalmatia. This region is recognized as an endemic region for canine visceral leishmaniosis, caused by Leishmania infantum. In November 2003, a 4-yr-old male gray wolf was found dead in the northwestern border of this endemic region.