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Open access

Mirosław Różycki, Ewa Chmurzyńska, Ewa Bilska-Zając, Jacek Karamon and Tomasz Cencek

Abstract

Introduction

Health, religious, and commercial aspects justify the need for meat species identification. The lack of officially approved methods prompts the undertaking of research on validation of isoelectric focusing of proteins (IEF) for official purposes.

Material and Methods

Samples were prepared from pigs (Sus scrofa ferus domestica), cattle (Bos taurus), and poultry (Gallus gallus domesticus). Meat mixtures were made by blending 50%, 25%, 10%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.2% meat of other species. Samples were examined on ultrathin polyacrylamide gels with pH 3–9 gradient.

Results

The results of the study confirmed the stable and reproducible pattern of meat protein bands. The detection limit of raw meat admixtures from pigs, cattle, and poultry mostly ranged from 2% down to 0.2% (0.2% for poultry). However, the IEF method can be used to detect the addition of pig meat to bovine meat in an amount higher than 3%. At the significant mixture level (i.e at least 5% addition of meat of another species) IEF proves itself with 100% specificity, sensitivity, and accuracy.

Conclusion

The achieved detection limits provide a basis for recommending the IEF method for routine tests in laboratories detecting the species origin of meat.

Open access

Ewa Bilska-Zając, Mirosław Różycki, Ewa Chmurzyńska, Jacek Karamon, Jacek Sroka, Ewelina Antolak, Marek Próchniak and Tomasz Cencek

Abstract

Introduction: The paper describes identification of Trichinella species isolated from wild boars (Sus scrofa) in the most popular hunting region of the West Pomeranian Province of Poland.

Material and Methods: The Trichinella larvae were identified by digestion method. For species identification of the larvae, multiplex PCR was used according to the European Reference Laboratory for Parasites Multiplex PCR protocol. The results were confirmed by molecular amplification of 5S rDNA gene and sequence analysis.

Results: Prevalence of 0.54% Trichinella–positive wild boars in the West Pomeranian Province was recorded. Examination of the larvae showed the occurrence of T. spiralis in 79 %, T. britovi in 16.5 %, mixed infection with T. spiralis/T. britovi in 3.5%, and T. pseudospiralis in 1.0% of the boars.

Conclusion: This is the first record of wild boar infected with non-encapsulated larvae of T. pseudospiralis in Poland. The species is very difficult to determine, especially using trichinoscopic method. The discovery of the larvae in the animals which may be intended for human consumption confirms that digestion technique should be the only method used for the inspection of meat, especially that from wild boars..

Open access

Ewa Cisak, Angelina Wójcik-Fatla, Jacek Sroka, Violetta Zając, Ewa Bilska-Zając, Ewa Chmurzyńska and Jacek Dutkiewicz

Abstract

Serum samples from 123 cattle, 95 wild boars, and 43 deer (red deer, roe deer, and fallow deer) from the territory of eastern Poland were examined by the ELISA for the presence of specific antibodies against tick-borne encephalitis virus (TBEV). The rates of positive response in the animals were 4.1%, 16.8%, and 11.6%, respectively. Examination of 37 blood samples from deer with RT-PCR revealed only one positive result in a roe deer (2.7%). The relatively high serologic response rate in wild boars was due to a very high response rate (35.7%) in the Chełm district, which accounted for 94% of the total positive results. These findings seem to indicate that the Chełm district is most probably an endemic area of TBEV.

Open access

Jacek Karamon, Maciej Kochanowski, Joanna Dąbrowska, Jacek Sroka, Mirosław Różycki, Ewa Bilska-Zając and Tomasz Cencek

Abstract

The aim of the study was to estimate the current prevalence of E. multilocularis in selected populations of red foxes in Poland and to evaluate the changes in prevalence of this parasite by comparison with the results obtained in the same area during earlier surveillance. The investigations were performed in the area of four Polish provinces: 2 eastern/south-eastern (Lubelskie and Podkarpackie) and 2 south-western (Śląskie and Opolskie). Five hundred red foxes coming from the investigated areas were examined between 2013 and 2014 to estimate the current situation in selected provinces. Moreover, 550 red foxes from the same areas examined between 2007 and 2013 were used for comparison of differences in E. multilocularis prevalences in time. Intestines were examined with the use of the sedimentation and counting technique. Among 500 foxes examined in the current study, 118 were positive for E. multilocularis. There were differences in prevalence between individual provinces: Podkarpackie Province - 54.6%, Lubelskie Province - 18.9%, Śląskie Province - 11.7%, and Opolskie Province - 3.9%. Statistical analysis demonstrated that in most cases there were no differences in prevalence between the current results and the results from previous studies. Only in Opolskie Province was a statistically significant increase observed between 2010 and 2014. A stable degree of infection in the region with high prevalence of this parasite was demonstrated. However, a significant increase in the region with very low prevalence of E. multilocularis points out the necessity to monitor this infection during the coming years to control the progress of the disease

Open access

Angelina Wójcik-Fatla, Jacek Sroka, Violetta Zając, Jacek Zwoliński, Anna Sawczyn-Domańska, Anna Kloc, Ewa Bilska-Zając, Robert Chmura and Jacek Dutkiewicz

Abstract

Introduction

Exposure to zoonotic factors in veterinary practice is closely related to the nature of the work. The main aim of the study was to determine the risk of selected zoonotic infections among the occupational group of veterinarians in Poland.

Material and Methods

Blood samples of 373 veterinarians (162 males and 211 females) from 12 provinces of Poland were collected by the venipuncture of a forearm for serological tests. Commercial immunoenzymatic tests (ELISA) were used for detection of specific IgG antibodies to Echinococcus granulosus, IgM and IgG to Leptospira spp., and IgM, IgA, and I and II phase IgG to Coxiella burnetii. Enzyme-linked fluorescence assays (ELFA) were used to detect IgM and IgG antibodies to Toxoplasma gondii.

Results

Positive results were found in 209 (56.0%) veterinarians for at least one of the examined diseases. The overall proportion of participants found to have specific Toxoplasma gondii antibodies in the IgM and/or IgG assays amounted to 44.5%. The presence of Coxiella burnetii antibodies was found in 16 (4.3%) subjects, while Leptospira spp. antibodies were detected in 63 (16.9%) veterinarians. Among the 373 veterinarians examined, no Echinococcus granulosus antibodies were found.

Conclusion

Results of the study seem to indicate a slightly elevated risk of Toxoplasma gondii infection and a moderate risk of infection with Leptospira spp. and Coxiella burnetii in veterinarians.

Open access

Jacek Karamon, Jacek Sroka, Tomasz Cencek, Mirosław Różycki, Ewa Chmurzyńska, Ewa Bilska-Zając, Jolanta Zdybel, Piotr Nowak, Jolanta Kędzierska and Piotr Dębiak

Abstract

The aim of the study was to optimise selected PCR methods for identification of T. solium, and to compare their effectiveness and usefulness. The investigation concerned three PCR methods described earlier: PCR I (specific to oncospherespecific protein Tso31 gene), PCR II (specific to large subunit rRNA gene), and PCR III (cytochrome c oxidases ubunit 1 gene). Each of them needed optimisation in connection with some changes in the procedures. Among the examined procedures, PCR I was found to be the most useful, requiring the least corrections during optimisation - only a higher concentration of polymerase was necessary. Testing an optimised PCR II method showed strong unspecific reactions with E. granulosus and T. saginata. This method was not considered diagnostically useful in distinguishing T. solium. PCR III method yielded products only when annealing temperature was lowered by 2 C. Under such conditions, there were no unspecific reactions with three others Taenidae parasites; however, annealing at a temperature only 1oC lower generated a distinct unspecific PCR product from T. saginata DNA. Therefore, this method was of limited usefulness. Comparison of the effectiveness of the two selected methods (PCR I and III) in detection of T. solium in successive DNA dilutions showed a large difference between them: in the same DNA sample, PCR I showed positive results in a sample diluted 1:3200, while PCR III failed at dilutions greater than 1:50. The results showed that among the three different methods used in the investigations, the most specific and effective for identification of T. solium was PCR I.

Open access

Jacek Karamon, Małgorzata Samorek-Pieróg, Bożena Moskwa, Mirosław Różycki, Ewa Bilska-Zając, Jolanta Zdybel and Magdalena Włodarczyk

Abstract

Introduction: The aim of the study was to determine the prevalence of intestinal helminths in red foxes (Vulpes vulpes) and raccoon dogs (Nyctereutes procyonoides) in the Augustów Primeval Forest (north-eastern Poland), with particular regard to zoonotic parasites.

Material and Methods: Intestines from 53 raccoon dogs and 66 red foxes were examined with the use of sedimentation and counting technique (SCT). Samples of faeces from 51 red foxes and 50 raccoon dogs were examined with the use of flotation method.

Results: Parasitic helminths were found by SCT in 98.5% of red foxes and 96.2% of raccoon dogs. Both species were infected with: Alaria alata (93.9% and 94.3%, respectively), hookworms (68.2% and 83.0%), Apophallus spp. (7.6% and 15.1%), Mesocestoides spp. (57.6% and 24.5%), Taenia spp. (40.9% and 1.9%), and Toxocara/Toxascaris nematodes (33.3% 15.1%). Echinococcus multilocularis was detected only in red foxes (6.1%), but trematodes Echinostomatidae and nematodes Molineus spp. only in raccoon dogs (18.9% and 41.5%, respectively). Additionally, Capillaria spp. eggs were detected by flotation method in 78.4% of foxes and 20.0% of raccoon dogs.

Conclusion: The study showed a very high percentage of red foxes and raccoon dogs infected with intestinal helminths in the Augustów Primeval Forest. Moreover, dangerous zoonotic parasites also were found, which should be taken into consideration in the assessment of infection risk for humans in this region.

Open access

Jacek Sroka, Zygmunt Giżejewski, Angelina Wójcik-Fatla, Krzysztof Stojecki, Ewa Bilska-Zając, Jacek Dutkiewicz, Tomasz Cencek, Jacek Karamon, Violetta Zając, Paweł Kusyk, Joanna Dąbrowska and Maciej Kochanowski

Abstract

The purpose of this study was to assess the possible influence of beavers on the contamination of lake water with zoonotic parasites Giardia duodenalis and Cryptosporidium spp., with respect to the risk to human health. A total of 79 water samples were taken around the habitats of beavers from 14 localities situated in the recreational Masurian Lake District (north-eastern Poland). Water was sampled in the spring and autumn seasons, at different distances from beavers’ lodges (0-2, 10, 30, and 50 m). The samples were examined for the presence of (oo)cysts of zoonotic protozoa Giardia duodenalis and Cryptosporidium spp. by direct fluorescence assay (DFA) and by nested and real time PCR. By DFA, the presence of Giardia cysts was found in 36 samples (45.6%) and the presence of Cryptosporidium oocysts in 26 samples (32.9%). Numbers of Giardia cysts, Cryptosporidium oocysts, and summarised (oo)cysts of both parasites showed a significant variation depending on locality. The numbers of Giardia cysts significantly decreased with the distance from beavers’ lodges while the numbers of Cryptosporidium oocysts did not show such dependence. The amount of Giardia cysts in samples collected in spring was approximately 3 times higher than in autumn. Conversely, a larger number of Cryptosporidium oocysts were detected in samples collected in autumn than in spring. By PCR, Giardia DNA was found in 38 samples (48.1%) whereas DNA of Cryptosporidium was found in only 7 samples (8.9%). Eleven Giardia isolates were subjected to phylogenetic analysis by restriction fragment length polymorphism PCR or sequencing which evidenced their belonging to zoonotic assemblages: A (3 isolates) and B (8 isolates). In conclusion, water in the vicinity of beavers’ lodges in the tested region was markedly contaminated with (oo)cysts of Giardia duodenalis and Cryptosporidium spp., which confirms the potential role of beavers as a reservoir of these parasites and indicates a need for implementation of appropriate preventive measures to protect tourists’ health.