Bożena Moskwa

The first detection of Neospora caninum DNA in the colostrum of infected cows

Limited data is available on the vertical transmission of Neospora caninum via the colostrum. The results of our previous research revealed the presence of N. caninum DNA in the milk of seropositive cows. The aim of the present work is to demonstrate parasite DNA in colostrum samples. A polymerase chain reaction using Np21 and Np6 primers was applied to DNA isolated from the colostrum sediment in order to amplify the corresponding genomic Nc-5 region. The expected 328-bp product was obtained in colostrum samples collected both on the calving day and the day after. This is the first detection of N. caninum DNA in the colostrum of seropositive cows, and these findings implicate the possibility of N. caninum transmission through the colostrum.

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.

Trichinella spiralis: in vitro cytotoxicity of peritoneal cells against synchronous newborn larvae of different age

Abstract Two different immune sera obtained from mice infected with muscle larvae (ML) of Trichinella spiralis (oral infection) or injected with 20-h-old newborn larvae (NBL) via retro-orbital venous plexus (intraocular injection) were compared in antibody-dependent cellular cytotoxicity test. Four synchronous stages of NBL, 0–2 h old, 22–24 h old, 46–48 h old and 6 days old (sNBL) were used to study susceptibility of larvae to the cytotoxic reaction of peritoneal cells. Peritoneal cells adhered to and destroyed sNBL of T. spiralis only in the presence of both immune sera. Living sNBL, living sNBL covered by peritoneal cells and dead sNBL were observed in all experimental groups. The lowest percentage of dead sNBL was detected in the 22- to 24-h-old groups. A dramatic increase in mortality was observed in older groups. A greater susceptibility of sNBL to serum obtained from mice infected per os with ML was observed. Serum obtained after oral infection showed higher levels of IgG1 isotype specific to ML than serum obtained after ocular injection.

First report of Trichinella pseudospiralis in Poland, in red foxes (Vulpes vulpes)

Nematode worms of the genus Trichinella are one of the most widespread zoonotic pathogens. Natural transmission between hosts can only occur through the ingestion of infected meat. To date, two Trichinella species are known to be etiological agents of disease among domestic animals and wildlife in Poland: T. spiralis and T. britovi. In the last decades, since the administration of an oral vaccination against rabies, the red fox population in Poland has increased exponentially. The study area covers the Nowy Targ region: a mountainous area (585–1138 m above the sea) in southern Poland. Of 24 red foxes examined in the study, four were infected with Trichinella isolates: three were identified as T. britovi and one as T. pseudospiralis. The muscle of red foxes infected with T. britovi harboured 2.75, 3.11, 4.4 LPG and with T. pseudospiralis 0.36 LPG. Trichinella larvae were identified at species level by genomic and mitochondrial multiplex PCR, the products of which were sequenced for comparison with other sequences available in GenBank. The sequences obtained from the Polish T. pseudospiralis isolate, deposited in GenBank under the accession numbers JQ809660.1 and JQ809661.1, matched sequences already published in GenBank. Sequence comparison showed a 100% match with the large subunit ribosomal RNA gene of T. pseudospiralis isolate ISS 013, and a 96–95% match with those of T. pseudospiralis isolates ISS 141 and ISS 470. This is the first report of the identification of T. pseudospiralis larvae from red fox in Poland.

In vitro isolation and identification of the first Neospora caninum isolate from European bison (Bison bonasus bonasus L.).

Peripheral blood from European bison (Bison bonasus bonasus L.) living in Białowieża Forest, north-east Poland, were investigated for the presence of antibodies to Neospora caninum and isolation of parasite. Out of 23 animals three of them showed a strong positive response to N. caninum (13%). The white blood cells from two positive and two negative bison were loaded on monolayer Vero cells culture. The first viable tachyzoites were detected only in positive samples at days 60 and 70 after incubation. For the purpose of the identification the isolates, tachyzoites were evaluated by PCR and sequence analysis of fragment of the Nc-5 region. The sequences of N. caninum-specific Nc5 region were found to be identical to those of other N. caninum isolates found in the public database. The isolate was subsequently named NC-PolBb1 and NC-PolBb2. This is the first record of in vitro isolation of N. caninum from naturally infected European bison (Bison bonasus bonasus L.) and the first isolate obtained from the peripheral blood.

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.

Detection of specific antibodies anti-Neospora caninum in the fallow deer (Dama dama).

Sera from 335 farmed fallow deer (Dama dama) at the breeding station in Kosewo Górne in the Mazurian Lake District, North-East Poland, were investigated for the presence of antibodies against Neospora caninum. The distribution of age groups was as follow: >4 years - 154 animals; 2 years - 76 animals; 1 year - 105 animals. Ten sera with the optical density exceeding 0.159 absorbance units (i.e., cut-off value) in ELISA test were also analyzed by Western blot. Western blot analysis revealed seroreactivity against immunodominant N. caninum antigens of 37, 25, and 16kDa; however, in some sera additional bands were also visible. This is the first screening studies for antibodies against N. caninum in farmed fallow deer in Poland, in the region where neosporosis was confirmed in cattle and in farmed and free-ranging European red deer (Cervus elaphus).

The repeatability of faecal egg counts in Polish Wrzosówka sheep

The number of nematode eggs in the faeces was estimated in Polish Wrzosówka sheep, in the spring and autumn of 1993 and again in 1994. The sheep had been naturally infected. The dominant species were Haemonchus contortus and Teladorsagia circumcincta, but Trichostrongylus spp., Cooperia curticei, Nematodirus spp. and Chabertia ovina were also present. Anthelmintics were not used. Egg counts were skewed, with a range of 0-4100 eggs g-1 (EPG); most sheep had egg counts below 100 EPG. Egg counts were approximately four times higher in spring than in autumn. Repeatability values within a season were all significant and positive. The repeatability of egg counts between seasons was estimated from the correlation between the mean transformed value in spring and in autumn, and was 0.52 in 1993 (P < 0.001) and 0.41 in 1994 (P < 0.05). The results show that animals with higher than average values in spring are likely to have higher than average values in autumn, and suggest that similar mechanisms regulated egg counts in both seasons even though egg counts were much lower in autumn.

The comparison of different ELISA procedures in detecting anti-Trichinella IgG in human infections.

The aim of this study was to compare the performance of four different ELISA procedures in diagnosing Trichinella spp. infections in human outbreaks. The main differences between the various procedures were: antigen concentration, dilutions of human serum sample, the type of conjugate used, and time of conjugate incubation. Serum samples were collected from 23 individuals involved in an outbreak. Results obtained in the four tested ELISA procedures differed in the rate of positive results obtained for the examined outbreak. Western blot confirmed the ELISA-positive results. Reactions to the specific 41-45kDa and 55kDa bands were obtained with all positive sera.