Anna M. Pyziel

The Annual Cycle of Shedding Eimeria Oocysts by European Bison (Bison bonasus) in the Bialowieza Primeval Forest, Poland

abstract:  Between April 2008 and March 2009, we analyzed the pattern of coccidian oocysts present in the feces of the European bison (Bison bonasus L., 1758) and found 4 species (Eimeria bovis, E. canadensis, E. ellipsoidalis, E. zuernii) previously reported from this host and 3 species (Eimeria alabamensis, E. cylindrica, E. pellita) that are new host and locality records. All the species occurred in bison females, and only 4 occurred in males; E. bovis was the most prevalent in both sexes. The overall prevalence of Eimeria spp. invasion reached 34.7% in cows and 13.9% in bulls. The highest prevalence was noted in early spring, with a peak in April, and the lowest in late autumn and winter. The oocyst count per gram of feces (OPG) varied from 50 to 1,350; no symptoms of clinical coccidiosis were observed. We found a significant influence of winter aggregations of bison on shedding of coccidian oocysts. The prevalence and OPG values were higher in bison congregating in large numbers around winter-feeding sites in comparison to other sites. We suggest that the coming together of cows during the growing season impacts the gender-related differences in prevalence and the number of coccidian species involved. This observation probably results from an increased production of oocysts by sub-clinically infected individuals in high-density bison populations.

The parasitic fauna of the European bison (Bison bonasus) (Linnaeus, 1758) and their impact on the conservation. Part 1 The summarising list of parasites noted

During the current century, 88 species of parasites have been recorded in Bison bonasus. These are 22 species of protozoa (Trypanosoma wrublewskii, T. theileri, Giardia sp., Sarcocystis cruzi, S. hirsuta, S. hominis, S. fusiformis, Neospora caninum, Toxoplasma gondii, Cryptosporidium sp., Eimeria cylindrica, E. subspherica, E. bovis, E. zuernii, E. canadensis, E. ellipsoidalis, E. alabamensis, E. bukidnonensis, E. auburnensis, E. pellita, E. brasiliensis, Babesia divergens), 4 trematodes species (Dicrocoelium dendriticum, Fasciola hepatica, Parafasciolopsis fasciolaemorpha, Paramphistomum cervi), 4 cestodes species (Taenia hydatigena larvae, Moniezia benedeni, M. expansa, Moniezia sp.), 43 nematodes species (Bunostomum trigonocephalum, B. phlebotomum, Chabertia ovina, Oesophagostomum radiatum, O. venulosum, Dictyocaulus filaria, D.viviparus, Nematodirella alcidis, Nematodirus europaeus, N. helvetianus, N. roscidus, N. filicollis, N. spathiger, Cooperia oncophora, C. pectinata, C. punctata, C. surnabada, Haemonchus contortus, Mazamastrongylus dagestanicus, Ostertagia lyrata, O. ostertagi, O. antipini, O. leptospicularis, O. kolchida, O. circumcincta, O. trifurcata, Spiculopteragia boehmi, S. mathevossiani, S. asymmetrica, Trichostrongylus axei, T. askivali, T. capricola, T. vitrinus, Ashworthius sidemi, Onchocerca lienalis, O. gutturosa, Setaria labiatopapillosa, Gongylonema pulchrum, Thelazia gulosa, T. skrjabini, T. rhodesi, Aonchotheca bilobata, Trichuris ovis), 7 mites (Demodex bisonianus, D. bovis, Demodex sp., Chorioptes bovis, Psoroptes equi, P. ovis, Sarcoptes scabiei), 4 Ixodidae ticks (Ixodes ricinus, I. persulcatus, I. hexagonus, Dermacentor reticulatus), 1 Mallophaga species (Bisonicola sedecimdecembrii), 1 Anoplura (Haematopinus eurysternus), and 2 Hippoboscidae flies (Lipoptena cervi, Melophagus ovinus). There are few monoxenous parasites, many typical for cattle and many newly acquired from Cervidae.

The first report of Angiostrongylus vasorum (Nematoda; Metastrongyloidea) in Poland, in red foxes (Vulpes vulpes)

Angiostrongylus vasorum belongs to the superfamily of Metastrongyloidea. This nematode occurs in foxes, dogs and other predators. The Nematode A. vasorum place themselves in the pulmonary artery and its branches, and in the right ventricle and atrium of the heart. Numerous species of land snails are the intermediate hosts of the parasite. In 2013, lungs and hearts of 76 foxes shot in the Forest District Głęboki Bród in Augustowska Primeval Forest were parasitologically necropsied. Four of the examined foxes were infected with the nematode A. vasorum, a prevalence of 5.2%. In one fox pericardium there were 6 male and 6 female nematodes. In the remaining three foxes nematodes were localized in the pulmonary artery. In two foxes 2 specimens of nematodes were detected (male and female, and two females) while 1 female was detected in the other fox. This is the first report of the presence of the nematode A. vasorum in fox in Poland.

The parasitic fauna of the European bison (Bison bonasus) (Linnaeus, 1758) and their impact on the conservation. Part 2 The structure and changes over time

During the last century the recorded parasite fauna of Bison bonasus includes 88 species. These are 22 species of protozoa, 4 trematode species, 4 cestode species, 43 nematode species, 7 mites, 4 Ixodidae ticks, 1 Mallophaga species, 1 Anoplura, and 2 Hippoboscidae flies. There are few monoxenous parasites, the majority of parasites are typical for other Bovidae and Cervidae species and many are newly acquired from Cervidae. This is an evident increased trend in the parasite species richness, in both the prevalence and intensity of infections, which is associated with the bison population size, host status (captive breeding or free-ranging) and the possibility of contact with other ruminant species. In light of the changes to parasite species richness during the last decades, special emphasis shall be given to new parasite species reported in European bison, their pathogenicity and potential implications for conservation.

The first detection of nematodes Ashworthius sidemi in elk Alces alces (L.) in Poland and remarks of ashworthiosis foci limitations

Ashworthius sidemi, a nematode belonging to the family of Trichostrongylidae, is a primary parasite of the Asian deer, mainly sika deer (Cervus nippon), with which it was introduced to Ukraine, as well as Slovakia, the Czech Republic and France. Migrating red deer carried this parasite from neighboring countries to Polish territory. Until now, in Poland, this parasite has been recorded in European bison, red deer, roe deer and fallow deer. As a result of post-mortem examinations of 10 elk, 2 of them from the Augustów Forest and Biebrza Marshes, A. sidemi were found in abomasa for the first time in Poland. The intensity of the invasions was 120 and 7 specimens, respectively. This finding of Ashworthius sidemi in elk indicates a further expansion of the focus of ashworthiosis in BiaŁowieża towards the north into the Biebrza Marshes and the Augustowska Forest. The growth of the elk population and their tendency for long distance migrations can contribute to the spread of the parasitosis in much greater distances than deer. On the basis of our own research and data from the literature, the current spread of ashworthiosis in Poland is discussed.

The prevalence and distribution of Dirofilaria repens in dogs in the Mazovian Province of Central-Eastern Poland

The aim of this study was to determine the prevalence and distribution of canine dirofilariosis from different districts of the Mazovian Province in central-eastern Poland. 462 dogs aged from 1.5-14 years were examined for dirofilariosis. Blood samples were examined for the presence of microfilariae using the Knott method, as well as the method of Kingston and Morton, after centrifugation in haematocrit microtubes in order to determine the intensity of infection as the number of microfilariae in 60 microliters of blood. The species of microfilariae found were determined after staining on the basis of the morphological characters. Samples were also examined using Canine Heartworm Antigen Test Kit SNAP HTWM (IDEXX, USA) that allows the detection of circulating antigens of females of Dirofilaria immitis. The positive samples were examined using multiplex PCR assay for species confirmation. Microfilariae belonging to the species D. repens were found in the blood samples of dogs originating from the city of Warsaw and from 18 districts of Mazovian Province. The mean prevalence of that species observed in the province was 25.8%. The range of intensity counted with the number of microfilariae found in 60 microliters of blood amounted to between 1-150; median intensity 9 microfilariae. The highest prevalence, reaching 52.9% and the highest median intensity of infection reaching 28 microfilariae, was found in Radom district. The lowest prevalence, amounting to 4.2%, was registered in Grójec district; however, the lowest median intensity reaching 6 microfilariae was noted in Żyrardów district. The infected dogs had not been outside of Poland, which means that this is an autochthonous infection. Microfilariae and antigens of D. immitis were not found in any examined blood samples. Results of the presented study show that autochthonous dirofilariosis caused by nematodes D. repens commonly occurs in the area of the Mazovian Province.

Immune responses in rats and sheep induced by a DNA vaccine containing the phosphoglycerate kinase gene of Fasciola hepatica and liver fluke infection.

Immune responses of rats and sheep following vaccination with cDNA encoding phosphoglycerate kinase of Fasciola hepatica (cDNA-FhPGK/pCMV) and F. hepatica infection were investigated in the present study. cDNA-FhPGK/pCMV vaccinated female Sprague-Dawley rats were better protected by vaccination than their male counterparts–48% reduction in fluke burden for females and no protection for males when compared with appropriate infection control groups. Moreover, male rats developed marked leukocytosis during the study with higher neutrophil, eosinophil and monocyte responses than females. Additionally, dynamics of eosinophil and monocyte responses varied between sexes. Increased titres of anti-FhPGK IgG1 and IgG2a correlated with the protective effect of vaccination that was observed among female rats. In the case of male sheep, no differences in worm burdens and in the course of the immune response were observed following vaccination. Titres of specific antibodies detected were low, and cellular responses were not significant. Apparently, sheep immune responses induced by cDNA-FhPGK/pCMV vaccination are not effective at controlling F. hepatica infection. Poor immunogenicity of DNA vaccines in large animals is still a major obstacle of this technology that has to be overcome.

Prevalence of Dirofilaria repens in dogs in central-eastern Poland and histopathological changes caused by this infection

Abstract The aim of the study was to determine the prevalence of canine dirofilariosis in four provinces of central-eastern Poland and to describe pathological changes occurring during the course of this parasitosis. A total of 188 dogs aged from one and a half up to 16 years were examined for dirofilariosis. Blood samples were analysed for the presence of microfilariae using Knott method, as well as the method of Kingston and Morton. Pathomorphological examination of two dogs was also performed. Histological sections were stained with H&E, AB-PAS, van Gieson, and von Kossa methods. Microfilariae of Dirofilaria repens were found in blood samples of dogs that originated from the Mazowieckie, Lubelskie, and Podlaskie provinces. The mean prevalence of that species was 12.7%. Autopsy revealed the adult nematodes in the subcutaneous connective tissue. Microscopic examinations indicated the presence of microfilariae in the liver, kidneys, lungs, heart muscle, and intestines. Necrotic foci, thrombosis, and infiltrations of different intensity with eosinophils in internal organs were also observed. Significant lesions in different internal organs together with the presence of numerous microfilariae suggest their important role in the disease process.

Molecular analysis of lungworms from European bison (Bison bonasus) on the basis of small subunit ribosomal RNA gene (SSU)

Dictyocaulosis (Nematoda: Trichostrongyloidea) is a widespread parasitosis of the European bison (Bison bonasus) inhabiting Bialowieza Primeval Forest. Bearing in mind the current coexistence of bison with wild cervids, and with domestic ruminants in the 19th and 20th century, the need arose for molecular identification of lungworm species. Molecular analysis was done on adult lungworms that were obtained from the respiratory track of four free-roaming bison euthanized as a part of the population health control program. As the result of the study four identical small subunit-ribosomal RNA gene sequences from the lungworms were obtained and deposited in GenBank as sequence, 1708 bp long (GenBank KC771250). Comparative analysis of the SSU rRNA sequences revealed the European bison to be a host for the bovine lungworm Dictyocaulus viviparus.

Interrelationships of Dictyocaulus spp. in Wild Ruminants with Morphological Description of Dictyocaulus cervi n. sp. (Nematoda: Trichostrongyloidea) from Red Deer, Cervus elaphus

Abstract Lungworms from the genus Dictyocaulus cause parasitic bronchitis (dictyocaulosis) characterized by coughing and severe lung pathology in both domestic and wild ruminants. In this study we investigated the interrelationships of Dictyocaulus spp. from European bison (Bison bonasus L.), roe deer (Capreolus capreolus), and red deer (Cervus elaphus) by nucleotide sequence analysis spanning the 18S RNA gene (small subunit [SSU]) and internal transcribed spacer 2 (ITS2) regions of the ribosomal gene array as well as the mitochondrial cytochrome c oxidase subunit 1 (cox1). Molecular analyses of sequence data obtained partly with novel primers from between 10 and 50 specimens from each host were carried out. Bayesian inference analysis revealed that each host species was infected with different genotypes. Analysis of cox1 sequence data showed a diverse genetic background and high evolutionary potential of Dictyocaulus taxa. Data from lungworms of European bison revealed a distinct genotype of Dictyocaulus viviparus, whereas Dictyocaulus capreolus was only found in roe deer. In contrast, red deer were infected with a taxon with unique SSU, ITS2, and cox1 sequences. These results indicate the occurrence of a novel genotype from red deer, which differs significantly from the National Center for Biotechnology Information reference sequence of Dictyocaulus eckerti. The molecular evidence was consistent with a morphological study with description and imaging of Dictyocaulus cervi n. sp. recovered from red deer. Dictyocaulus cervi n. sp. can be distinguished from D. eckerti on the basis of the absence of cervical papillae, the occurrence of a single ring of 4 symmetrical submedian cephalic papillae, length of the tail in females, morphometry of the female reproductive system, and measurements of gubernacula in males. In conclusion, our findings further strengthen the idea that the genetic complexity and diversity among Dictyocaulus lungworms infecting wildlife ruminants is larger than previously believed and warrants further investigation.