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Open access

Jacek Karamon, Jacek Sroka, Tomasz Cencek, Maciej Kochanowski and Joanna Dąbrowska

Abstract

The aim of the study was to estimate the effectiveness of intestinal scraping technique (IST) in the detection of Echinococcus multilocularis. The experimental assessment of the limit of detection and comparison with “gold standard” (sedimentation and counting technique - SCT) was also performed. Samples of fox small intestines experimentally enriched with known numbers of E. multilocularis tapeworms, were used. Twenty four samples containing 10, 30, 60, and 90 E. multilocularis tapeworms were prepared. Moreover, in order to compare IST with SCT, 127 intestines of foxes were examined using both methods. The limit of detection was estimated at 30 E. multilocularis tapeworms per sample of the intestine. Moreover, mean number of Echinococcus found by IST were several dozen times lower than the real content of these tapeworms in the samples (on average only 2 to 3.2% of worms were recovered). Among 127 intestinal samples examined with the use of two methods, eight samples (8.2%) were positive by SCT and only two (1.6%) when IST was used. A relatively high limit of detection estimated experimentally in the first part of the study, as well as, the results obtained in field investigations showed clearly that IST method could significantly decrease the reliability of the results of investigations, especially carried out in regions where a very low prevalence of E. multilocularis occurs or in countries, which want to demonstrate that they are free from this parasite.

Open access

Jolanta Zdybel, Tomasz Cencek, Jacek Karamon and Teresa Kłapeć

Abstract

The objective of the study was to determine the degree of municipal wastewater contamination with intestinal parasite eggs of the genera Ascaris, Toxocara, and Trichuris at individual stages of treatment, and indication of potentially weak points in the hygienisation of sewage sludge. The study was conducted in 17 municipal mechanical-biological wastewater treatment plants which, to a slight degree, differed in the technological process of wastewater treatment and the method of hygienisation of sewage sludge. The selected treatment plants, located in seven regions, included five classified as large agglomerations (population equivalent - PE >100 000), ten as medium-size (PE 15 000-100 000), and two as smaller size with PE 10 000 - 5000. The largest number of viable eggs of Ascaris spp., Toxocara spp., and Trichuris spp. was found in the sewage sludge collected from the primary settling tank. A slightly lower number of the eggs were found in the samples of excess sludge, which indicates that the sedimentation process in the primary settling tank is not sufficiently long to effectively separate parasites’ eggs from the sewage treated. The number of eggs of Ascaris spp. and Toxocara spp. in the fermented sludge was nearly 3 times lower than that in the raw sludge. The effectiveness of hygienisation of dehydrated sewage sludge by means of quicklime was confirmed in two wastewater treatment plants, with respect to Ascaris spp. eggs, in three plants with respect to Toxocara spp. eggs, and in one plant with respect to Trichuris spp. eggs. The mean reduction of the number of eggs was 65%, 61%, and 100%, respectively. In one wastewater treatment plant, a reduction in the number of viable eggs of Ascaris and Trichuris species was also noted as a result of composting sludge by 85% and 75%, respectively. In the remaining treatment plants, no effect of hygienisation of sewage sludge was observed on the contents of viable eggs of these nematodes.

Open access

Małgorzata Samorek-Pieróg, Jacek Karamon and Tomasz Cencek

Abstract

Taenia solium is a parasite causing porcine cysticercosis and human taeniosis and cysticercosis, parasitic zoonoses with a serious public health and economic influence. It has been globally ranked as the top foodborne parasite by the Food and Agriculture Organisation of the United Nations (FAO) and the World Health Organisation (WHO). This parasite is transmitted mainly in countryside regions where animals are free roaming, having access to human faeces, and infected pork is widely available. More developed countries eliminated cysticercosis; nonetheless, there are insufficient data about the current endemicity status of T. solium, due to increased human migration from endemic areas. Formally submitted statistics on cysticercosis in pigs are extremely inadequate. This is the result of not reporting all cases of the disease by some countries and lack of molecular verification during identification of the parasite. There is a need to develop diagnostic tests with increased sensitivity and specificity. The purpose of the present review is to summarise current knowledge about diagnostic and control methods concerning T. solium infection. The article does not address the diagnostics of human cysticercosis, since there is a distinct medical field which should be discussed separately. The paper focuses mainly on identifying the sources of T. solium infection, presenting the methods to detect and control porcine cysticercosis and taeniosis in humans.

Open access

Tomasz Cencek, Jacek Karamon, Jacek Sroka and Jolanta Zdybel

Abstract

This article presents the essential stages leading to development of a modified method for Hypoderma bovis protein transfer, and estimation of the usefulness of semi-quantitative densitometric analysis of western-blottig results. The principal of the method was to incubate the gel obtained with the native electrophoresis method in buffer containing SDS prior to transfer of the separated proteins of L1 H. bovis onto nitrocellulose membrane. In two experiments, the authors estimated the efficacy of the new method and the possibility of using this method for semi-quantitative densitometric investigations.

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

Jacek Karamon, Maciej Kochanowski, Tomasz Cencek, Magdalena Bartoszewicz and Paweł Kusyk

Abstract

The aim of the study was to estimate the prevalence of gastrointestinal parasites in raccoons with particular regard to zoonotic parasites. Fifty-five raccoons, hunted or found dead on roads, were examined. The small and large intestines were collected from all raccoons and, additionally, the stomach was collected from 43 animals. The samples were examined with the use of sedimentation and counting technique. The intestines and stomach were examined separately. Samples of raccoon faeces were collected from their environment localised in Słubice district, Lubuskie province (Poland). The samples were collected once a month in 2012. In total, 154 faecal samples were obtained and examined with the use of McMaster flotation technique. The following parasites were detected in the intestinal and stomach contents: tapeworms Mesocestoides sp. (67.3%), Echinostomatidae flukes (34.5%), and nematodes Capillaria sp. (25.5%). Moreover, Acanthocephala were found in the intestines of three raccoons. The highest intensity of infection were observed in case of Mesocestoides sp. Mesocestoides sp. and Echinostomatidae were found statistically more often in the intestines than in the stomach. In the case of these two parasites, there was positive correlation between the intensity of infection in the intestines and the presence of the same parasites in the stomach. Moreover, significantly higher prevalence and intensity of Mesocestoides sp. in males than in females were also observed. Faecal samples contained Baylisascaris procyonis eggs (mean 60 epg). These eggs were found in three samples collected in November and December. Furthermore, in some faecal samples eggs of flukes, Capillaria sp., Mesocystoides sp., and coccidian oocysts were found. It is one of rare reports concerning Baylisascaris procyonis in Poland confirming the presence of this dangerous parasite in Polish raccoon population.

Open access

Maciej Kochanowski, Jacek Karamon, Joanna Dąbrowska, Arkadiusz Dors, Ewelina Czyżewska-Dors and Tomasz Cencek

Abstract

Introduction: The aim of study was to estimate the prevalence and intensity of intestinal parasite infections in pigs in Poland and evaluate the influence of factors related to the production system on the infection intensity.

Material and Methods: A total of 70 pig farms of all Polish provinces, differing in the herd size and production system, were selected for the study. Fresh faecal samples were collected from all age groups: suckling piglets, weaners, fatteners, and lactating sows. Moreover, data were obtained regarding the size of the herd, the use of paddock and all-in/all-out system, the presence of diarrhoea, and the type of flooring.

Results: Parasite eggs or oocysts were detected in 57 of the 70 examined pig farms. Oesphagostomum spp. eggs were found in the largest number of farms (68.6%). Moreover, coccidia (42.9%), Ascaris suum (28.6%), Trichuris suis (21.4%), and Strongyloides spp. (11.4%) were detected. The highest prevalence of coccidia and Strongyloides spp. was found in suckling piglets, A. suum and T. suis in fatteners, and Oesphagostomum spp. in sows. Higher prevalence of parasites was detected in small farms than in medium and large farms, except the prevalence of coccidia, which was the highest in medium farms. Simultaneous infection with several parasites was more often detected than with one parasite. Odds ratio of parasites occurrence was higher in farms with paddock and litter floor and in farms which do not use all-in/all-out system.

Conclusion: Relatively high prevalence of intestinal parasites was found in pigs in Poland. Moreover, specific distribution of parasites in different age groups and farms of different size was observed. Influence of breeding factors on parasite prevalence was identified.

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

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

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..