Prevalence and genetic characterization of Echinococcus spp. in cattle, sheep, and swine in Hungary.

A study was conducted to investigate genetic diversity of Echinococcus isolates collected in Hungarian slaughterhouses between 2015 and 2018. Organs of 219 animals with suspected hydatidosis were collected during routine meat inspection and sent to our laboratory. Echinococcus infection was confirmed in 65 animals. These results indicate that prevalence data based on reporting of slaughterhouses are not reliable without the appropriate training of the people responsible for meat inspection. The genetic diversity was evaluated by the DNA sequence analysis of the cytochrome c oxydase subunit 1 (cox1) mitochondrial gene. Echinococcus intermedius (n = 31), Echinococcus granulosus s.s. (n = 2), and Echinococcus multilocularis (n = 3) was identified in swine. In cattle, only E. granulosus s.s. (n = 20) was detected. E. granulosus s.s. (n = 7) was the dominant species in sheep; nevertheless, E. intermedius was also identified in two animals. E. granulosus s.s. (n = 29) and E. intermedius (n = 33) were classified in 13 and three haplotypes, respectively. The genetic diversity and haplotype network of E. granulosus s.s. were similar to that observed in some other countries of Eastern Europe. The genetic diversity of E. intermedius was low with a single dominant haplotype. Cysts were fertile in nine sheep (100%), 22 swine (61%), and three cattle (15%) indicating that all three species play a role in some extent in the epidemiology of cystic echinococcosis in Hungary. Based on the number of animals killed in the slaughterhouses involved in the present study, the rate of infection was 0.013% in sheep, 0.007% in cattle, and 0.001% in swine. As animals with hydatidosis originated from family farms, control programs should mainly focus on these facilities.

Environmental determinants of the spatial distribution of Echinococcus multilocularis in Hungary.

Human alveolar echinococcosis, caused by the metacestode stage of Echinococcus multilocularis, is one of the most pathogenic zoonoses in the temperate and arctic region of the Northern Hemisphere. To investigate the spatial distribution of E. multilocularis and the factors influencing this distribution in the recently identified endemic area of Hungary, 1612 red fox (Vulpes vulpes) carcasses were randomly collected from the whole Hungarian territory from November 2008 to February 2009 and from November 2012 to February 2013. The topographic positions of foxes were recorded in geographic information system database. The digitized home ranges and the vector data were used to calculate the altitude, mean annual temperature, annual precipitation, soil water retention, soil permeability, areas of land cover types and the presence and buffer zone of permanent water bodies within the fox territories. The intestinal mucosa from all the foxes was tested by sedimentation and counting technique. Multiple regression analysis was performed with environmental parameter values and E. multilocularis counts. The spatial distribution of the parasite was clumped. Based on statistical analysis, mean annual temperature and annual precipitation were the major determinants of the spatial distribution of E. multilocularis in Hungary. It can be attributed to the sensitivity of E. multilocularis eggs to high temperatures and desiccation. Although spreading and emergence of the parasite was observed in Hungary before 2009, the prevalence and intensity of infection did not change significantly between the two collection periods. It can be explained by the considerably lower annual precipitation before the second collection period.

Echinococcus multilocularis and Trichinella spiralis in golden jackals (Canis aureus) of Hungary.

Over the last decades the distribution area of the golden jackal (Canis aureus) has increased significantly in Europe, particularly in the Balkan Peninsula and in Central Europe. Vagrant individuals were described in many European countries. Herein, we report Echinococcus multilocularis (total worm count: 412) and Trichinella spiralis (101 larvae/g for muscles of the lower forelimb) infections in two golden jackals shot in Hungary. It is a new host record of E. multilocularis and T. spiralis in Europe and Hungary, respectively. As jackals migrate for long distances through natural ecological corridors (e.g., river valleys), they may play a significant role in the long distance spread of zoonotic parasites into non-endemic areas of Europe. Therefore, monitoring zoonotic parasites in this host species can be recommended in the European Union.

Spatial distribution and genetic diversity of Echinococcus multilocularis in Hungary.

Human alveolar echinococcosis, caused by the tapeworm Echinococcus multilocularis, is the most pathogenic helminthic zoonosis in the temperate and arctic region of Europe. Between November 2008 and February 2009, 840 red fox (Vulpes vulpes) carcasses, were randomly collected from the whole Hungarian territory. The intestinal mucosa from all the foxes was tested by sedimentation and counting technique. E. multilocularis adult worms were detected in foxes of 16 out of the 19 Hungarian counties and in the suburban areas of the capital, Budapest. The prevalence and abundance of infection was significantly (P<0.001) higher in the north-western half (16.2%, CI=14.5-17.9; m ± SE=165.5 ± 112.4) than in the south-eastern half of the country (4.2%, CI=3.2-5.2; m ± SE=3.6 ± 2.1). The highest prevalence (26.6%, CI=22.5-30.8%) and abundance (m ± SE=614.2 ± 469.3) was observed in the Northern Mountain Region bordering Slovakia. The multi-locus microsatellite analysis of 81 worms showed the presence of four out of the five main European profiles. The H profile was the most common profile (55.5%) with nine genotypes, followed by the G (18.5%) with two genotypes, E (13.6%) with one genotype and D (12.4%) with two genotypes. The genetic distance was not statistically correlated with the geographical distance of the samples, supporting the hypothesis that the geographical distance is only a minor factor among those involved in the genetic distribution of this parasite in Europe. These data indicate that Hungary should be considered as a peripheral area of a single European focus, where the dispersal movement of foxes resulted in the spreading of the parasite from one county to another within a time period short enough to avoid a substantial genetic drift.

Canine onchocercosis in Greece: report of further 20 cases and molecular characterization of the parasite and its Wolbachia endosymbiont.

Recently, sporadic cases of subconjunctival Onchocerca infection have been reported in dogs in Greece and Hungary. Herein we report further cases from Greece and the results of the molecular analysis of Onchocerca sp. removed from Greek dogs and its Wolbachia endosymbionts. Twenty dogs of various breeds, 1-11 years of age with subconjunctival onchocercosis (4 cases each in right or left eye, 12 cases in both eyes) were presented having similar manifestations. Periorbital swelling, exophthalmos, lacrimation, discharge, photophobia, conjunctival congestion, corneal edema, protrusion of the nictitating membrane, and subconjunctival granuloma or cyst formation were the most important clinical signs. After surgical excision of the periocular masses containing the worms, all animals recovered fully from onchocercosis. Based on the similarities of the clinical picture of the Greek and Hungarian cases, the similar morphology of the Greek and Hungarian isolates, and the identical sequences of the cytochrome oxidase gene of the filarial parasites and that of the 16S ribosomal RNA gene from their Wolbachia endosymbionts, the Onchocerca sp. isolated from dogs in Greece and Hungary appears to belong to the same species.

Characterization of Cryptosporidium spp.--recent developments and future needs.

Cryptosporidia, widely distributed protozoan parasites of vertebrates have recently attracted increasing interest due to several serious waterborne outbreaks, the life-threatening nature of infection in immunocompromised patients, and the realization of economic losses caused by these pathogens in livestock. Genetic polymorphism within Cryptosporidium spp. is being detected at a continuously growing rate, owing to the widespread use of modern molecular techniques. The aim of this paper is to review the current status of taxonomy, genotyping, molecular phylogeny, and characterization of cryptosporidia, and to highlight the need for polyphasic typing, i.e. an integrated approach comprising standardized morphologic, biologic, and molecular methods for describing Cryptosporidium species and isolates, and for establishing "virtual" reference strains.

Subconjunctival zoonotic onchocerciasis in man: aberrant infection with Onchocerca lupi?

In the past few decades, 10 cases of cryptic, zoonotic onchocerciasis, including two subconjunctival infections, have been reported in man. In the majority of cases, Onchocerca cervicalis, O. gutturosa or O. dewittei, which normally infect horses, cattle and wild boar, respectively, were responsible for the lesions. However, the taxonomic status of the parasites involved in the two subconjunctival infections, both of which were European, has never been unambiguously determined. In such infections, the acute phase appears to be characterized by conjunctivitis. A single, strongly coiled, immature, female worm was found incorporated in a large granulomatous nodule, in the ocular and peri-ocular tissues, in the chronic stage of each of the two eye infections. Several, patent, sporadic cases of subconjunctival O. lupi infection have recently been reported in dogs. In terms of the location of the worms, clinical signs and histopathology, these canine infections were very similar to those seen in the two human patients with eye infection. When the parasites recovered from human eyes were compared morphologically with the Onchocerca spp. infecting animals in Europe, they appeared to be most similar to O. lupi. Although O. lupi is normally a parasite of dogs, it may thus also be responsible for aberrant, zoonotic, subconjunctival infections in man.

Ocular onchocercosis in dogs: a review.

In recent decades, sporadic cases of ocular Onchocerca species infection have been reported in dogs in the USA and Europe. In the acute stage of the disease severe inflammation of the ocular and periocular tissues was observed. In chronic cases, the strongly coiled, gravid nematodes were incorporated in pea- to bean-sized granulomatous nodules in various parts of the eye, including the retrobulbar space, orbital fascia, eyelid, third palpebra, conjunctiva and sclera. Apart from the ophthalmological significance of the disease, the large number of microfilariae in the skin may be responsible for acute and chronic dermatological problems. The geographical distribution and prevalence of the infection may be greater than currently thought, because the lesions may have been erroneously regarded as other ocular diseases. Onchocerciasis is the world's second most prevalent infectious cause of blindness in human beings and parasitologists have long searched for an experimental model of human onchocerciasis; ocular onchocercosis infections in dogs may provide a useful experimental system.

Polyphasic typing of Cryptosporidium baileyi: a suggested model for characterization of cryptosporidia.

The present study was undertaken to characterize the oocyst morphology, host specificity, organ location, virulence, and sequences of the small subunit ribosomal RNA, 70-kDa heat shock protein, and oocyst wall protein genes of Cryptosporidium baileyi, and to compare this strain with other Cryptosporidium species. This study also aims to serve as a model for polyphasic (phenetic and genetic) characterization of Cryptosporidium species and strains. On the basis of these results, further genetic and phenetic characterization of an avian isolate is needed if the difference between the length or width, or both, of oocysts of an isolate and of C. baileyi is > or = 10% or if the difference between the oocyst shape index of the isolate and of C. baileyi is > or = 3% (or both). The isolate is infectious for mammals or lower vertebrates, or the host range is narrow, i.e., infectious only for some bird species; after oral or intratracheal inoculation, the parasites are not located in the cloaca and in the bursa of Fabricius or the respiratory tract; clinical disease or weight gain reduction can be observed after oral inoculation; the genetic distance for the examined gene between C. baileyi and the isolate is similar in magnitude to that observed between most closely related Cryptosporidium species.

Canine ocular onchocercosis in Hungary.

An adult male mongrel dog that had spent its entire life in Hungary, was found to have infection with filaroid nematodes of the genus Onchocerca. The gravid male and female parasites were embedded in bean-sized granulomatous masses on the conjunctiva and the sclera of both eyes. The cuticle of females consisted of two separated layers in longitudinal sections, the external layer bearing ridges and the internal layer showing striations. The ridges were marked, rounded in shape, and the ratio of body diameter to the distance between ridges varied between 7:1 and 10:1. At midbody of the worms, two striations could be seen between each pair of ridges: one under every ridge and one between neighbouring ridges. Numerous exceptionally small (96.4 microm x 6.4 microm) microfilariae were seen in the uteri of females and the surrounding tissues and isolated from skin biopsy materials. The morphology and location of the parasite and histopathological lesions of the Hungarian case were similar to that described in dogs in the United States. This case is the first documented ocular Onchocerca infection in dogs outside the western United States. Thus, onchocercosis should be considered in the differential diagnosis of ocular and periocular nodules in dogs also in Europe.