Echinococcus multilocularis genetic diversity based on isolates from pigs confirmed the characteristic haplotype distribution and the presence of the Asian-like haplotype in Central Europe.

Introduction: The aim of the study was to determine the genetic diversity of Echinococcus multilocularis in pigs in highly endemic areas in Poland, as well as to attempt to confirm the occurrence and geographical distribution of haplotypes characteristic for these areas, which were previously described on the basis of examination of adult tapeworms isolated from foxes. Material and Methods: Twenty samples of E. multilocularis larval forms were obtained from pigs' livers in four provinces of Poland. Genetic analyses were conducted on sequences of two mitochondrial genes: cox1 and nad2. Results: Seven haplotypes were found for the cox1 gene (OQ874673-OQ874679) and four haplotypes for nad2 (OQ884981-OQ884984). They corresponded to the haplotypes described earlier in foxes in Poland (some of them differing only in one nucleotide). The analysis showed the presence of the Asian-like haplotype in both the cox1 and nad2 genes. The remaining haplotypes were grouped in the European clade. The geographical distribution of haplotypes identified in the pig samples was noticed to bear a similarity to the distribution of haplotypes previously isolated from foxes in the same regions. Conclusion: The characteristic geographical distribution of E. multilocularis haplotypes in Central Europe (including the presence of the Asian-like haplotype) previously described in the population of definitive hosts (foxes) has now been confirmed by the analysis of samples from non-specific intermediate hosts (pigs).

Gelatin medium for preserving of Trichinella spp. for quality control in laboratories - estimation of larvae viability.

INTRODUCTION AND OBJECTIVE: Official food control laboratories ensure food safety using reliable, validated methods. Council Regulations (EC) No. 853/2004, 854/2004 and 882/2004 of the European Parliament established hygiene rules the production of food of animal origin, together with requirements for official controls. This leads to detailed requirements for Trichinella control set out in Commission Implementing Regulation (EU) 2015/1375 of 10 August 2015. These regulations require the laboratory to participate in proficiency testing (PT) to confirm their competence and improve the quality of testing, and require the PT Organizer to use methods for the preparation and preservation of parasite larvae in order to evaluate and improve detection. Traditional methods of preparing such larvae expose them to rapid degradation, making it necessary to simultaneously isolate the larvae and place them in meatballs to ensure quality. MATERIAL AND METHODS: We developed a technique for preserving of Trichinella spp. for quality control such as PT sample preparation. The procedure protects larvae against toxic oxygen activity and bacterial destruction via a gelatin barrier. To estimate the viability of larvae preserved by this method, gelatin capsules with 10 larvae of T. spiralis in each were stored (4-8 °C) during 45 days of an experiment. Samples were tested at 2 day intervals (3 samples each day of testing). RESULTS: In total, 75 samples were tested. Larvae remained alive up to 3 weeks. The number of living larvae diminished after 27 days through day 43, after which no living larvae were observed. CONCLUSIONS: The gelatin medium procedure facilitated easy, high-throughput sample preparation and supported 100% recovery for 3 weeks. The method allows fast, efficient and accurate PT sample preparation.

Molecular confirmation of the systemic toxoplasmosis in cat.

INTRODUCTION AND OBJECTIVE: Systemic toxoplasmosis with tissue-spread parasites occurring in intermediate hosts may also occur in immunocompromised cats (e.g., infected with FLV or FIV). To the best of our knowledge, no reports have been published on the detection and genotyping of T. gondii DNA in cats with extraintestinal toxoplasmosis in Poland. The article describes the case of the sudden death of 3 out of 4 cats in a cattery, and the detection and molecular characterization of T. gondii DNA detected in the tissues of one of the dead cats. MATERIAL AND METHODS: Samples of brain, lungs, heart, and liver of the cat that died suddenly were examined for the presence of T. gondii DNA (B1 gene) by nested PCR and real-time PCR. DNA positive samples were also genotyped at 12 genetic markers using multiplex multilocus nested PCR-RFLP (Mn-PCR-RFLP) and multilocus sequence typing (MLST). RESULTS: A total of 9 out of the 20 DNA samples were successfully amplified with nested and/or Real-time PCR. DNA from 3 out of 5 types of tested samples were genotyped (brain, heart and muscle). Mn-PCR-RFLP and MLST results revealed type II (and II/III at SAG1) alleles at almost all loci, except a clonal type I allele at the APICO locus. This profile corresponds to the ToxoDB#3 genotype, commonly identified amongst cats in Central Europe. CONCLUSIONS: To the best of our knowledge, this is the first study describing the genetic characteristics of T. gondii population determined in a cat in Poland. These data confirm the importance of this host as a reservoir for this pathogen, and demonstrate the genotypic variation of this parasite. Veterinarians should take into account that cats may develop disseminated toxoplasmosis, and that it is a systemic disease which may lead to the death of the cat, and to transmission of the pathogen to other domestic animals and to humans.

Parasitological contamination of arable soil in selected regions of Poland - preliminary study.

INTRODUCTION AND OBJECTIVE: The hygienic status of arable soils in most developed countries has been unknown. In the presented study, a preliminary investigation was undertaken to determine the contamination with eggs of parasitic nematodes in the soil of arable fields in Poland. The aim of the study was to determine whether such contamination is common enough to constitute a significant problem and what factors may influence it. MATERIAL AND METHODS: The study was conducted in 5 Polish provinces from autumn 2021 to spring 2022. The provinces differed significantly in terms of the area of agricultural land, agricultural suitability, type of soil, scale of cattle and pig breeding, production of manure and slurry, and the use of manures and organic fertilizers for fertilization. A total of 133 soil samples were collected. Parasitological examination of soil samples was carried out using the PN-Z-19006 method [1], with confirmed high sensitivity. RESULTS: Parasite eggs were found in a total of 67 samples, of which 56 samples contained eggs of roundworms of the genus Ascaris (an average of 3.29 eggs/100 g of soil), 23 contained eggs of whipworms (an average of 1.22 eggs/100 g), and 3 contained eggs of Toxocara (1 egg/100 g). CONCLUSIONS: Differences in the percentage of positive samples were found depending on the period in which the samples were taken. The percentage of positive samples collected in autumn (53.57%) was higher than the percentage of positive samples collected in spring (48.05%). Similarly, the average number of eggs of in positive samples collected in autumn (3.43 eggs/100 g) was higher than the average number of eggs in samples collected in spring (2.90 eggs/100 g). Differences in the percentage of positive samples were also found depending on the region of origin of the samples.

Comparison of the effectiveness of various parasitological methods in detecting nematode eggs in different types of soil.

INTRODUCTION AND OBJECTIVE: Natural fertilizers, sewage sludge, digestates, as well as organic fertilizers produced on their basis, can become a source of parasitological contamination of cultivated land. High concentration of invasive forms of parasites in the soil may pose a threat to human and animal health. Therefore, it is necessary to control the hygienic condition of fertilizers and fertilized soils with particular emphasis on parasites. The aim of the study was to compare the effectiveness of methods commonly used for parasitological examination of soil with own methods which were used to develop the standards. MATERIAL AND METHODS: The study was carried out using samples of sandy soil (SS), horticultural mix soil (HS) and peat-based substrate (PS). Each sample was spiked with 100 dyed Ascaris suum eggs and examined with the use of 6 methods: Vasilkova, Dada, Quinn, and 3 methods according to the Polish Standards (PN-19000, PN- 19005, PN-19006). For each variant, 8 repetitions were made. RESULTS: The largest number of A. suum eggs were found with PN-19006 (mean number of detected eggs was 21.25, 46.50, 23.00 for HS, SS, PS, respectively. Slightly lower results were obtained using PN-19005 - the mean number eggs was 21.25, 36.00, 16.75, respectively. On the other hand, the mean number of A. suum eggs found with the Dada method was about 2-3 times lower than with the PN-19006 - 15.75, 22.50, 6.50 for HS, SS, PS soil, respectively. Other methods were much less effective. CONCLUSIONS: PN-19006 method turned out to be the most effective in detecting A. suum eggs. This method can be used for parasitological examination of soils and can be the basis for developing a system of methods dedicated to testing different types of soils for the presence of nematode eggs.

Comparison Study of Four Extraction Methods Combined with PCR and LAMP for Feline Tritrichomonas foetus Detection in Fecal Samples.

Feline trichomonosis occurs worldwide, with gastrointestinal symptoms such as chronic large-bowel diarrhea and abdominal pain. The inclusion of molecular methods in diagnostic and epidemiological studies has necessitated an effective method for extracting DNA from feces. We tested four extraction commercial kits: ZR Fecal DNA MiniPrep (50 preps) (Zymo Research, Irvine, CA, USA), QIAamp® DNA Stool Mini Kit (Qiagen Inc., Valencia, CA, USA), UltraClean Fecal DNA Kit (50 preps) (MO BIO, San Diego, CA, USA), and Sherlock AX/100 isolations (A&A Biotechnology, Gdynia, Poland). We assessed the sensitivity of detection of Tritrichomonas foetus in spiked fecal samples for the four kits combined with two molecular assays: PCR and LAMP. The extraction efficacy was quantified using defined aliquots of fecal samples spiked with 5 µL of suspensions containing serial dilutions of trophozoites (0.1; 1; 10; 100; 1000; 10,000), with six replicates for each concentration. In our study, we proved that the ZR Fecal DNA MiniPrep (50 preps) kit combined with LAMP and PCR had the highest efficiency among all the compared methods for the detection of feline T. foetus from fecal samples.

Results of Proficiency Testing for Trichinella in Poland, 2015-2019.

Trichinellosis is a zoonotic meat-borne disease caused by the nematodes of the genus Trichinella. Meat containing live Trichinella larvae is a source of infection. The examination of meat for Trichinella was introduced in 1869, but the digestion method for this did not appear in Poland until the late 1970s. Nowadays, the meat of all food animals susceptible to Trichinella spp. is examined in the frame of official post mortem control with the digestion method. The majority of laboratories in Poland meet the requirements of the ISO/IEC 17025 Standard (352 field laboratories). Laboratory personnel participate in quality control programs. This paper presents the results of proficiency tests (PTs) organized within 2015-2019 in Poland. Over this period, the laboratories examined 7580 samples (contamination levels: zero, one, three, and five larvae). Each laboratory was provided with a set of samples (one negative and three positive). Over 95% of the samples were considered correct in qualitative assessments, though the results of the quantitative evaluations were slightly lower, with 89% of samples being considered correct. Based on a sample evaluation, 88% of laboratories passed the PT comparison. A slight decrease was observed in the examination of samples spiked with five larvae, and great progress was achieved in samples containing three larvae. Low levels of sample contamination are sought after in laboratories but may make evaluations difficult. For this reason, we must consider increasing the number of larvae added to the samples in the next PTs.

Molecular Confirmation of Massive Taenia pisiformis Cysticercosis in One Rabbit in Poland.

The aim of this study was to provide molecular characterization, together with phylogenetic analysis, of Taenia pisiformis cysts isolated from rabbit. On the basis of morphological features and molecular analysis, the cysticerci were identified as T.pisiformis metacestodes. PCR was performed with three different protocols to obtain partial sequences of 12S ribosomal RNA (12S rRNA), NADH dehydrogenase subunit 1 (nad1), and cytochrome oxidase subunit 1 (cox1) of Taenia spp. The products from the PCRs were sequenced. Interpretation of the sequencing results of the obtained amplicons, by comparing them with the GenBank database, proved that the causative agent, in this case, was T. pisiformis. The phylogenetic analysis of the received sequences identified a new haplotype. The received data can be used to supplement the species description. To our knowledge, this is the first molecular confirmation of T. pisiformis metacestodes infection in the rabbit, in Poland.

The First Record of Echinococcus ortleppi (G5) Tapeworms in Grey Wolf (Canis lupus).

The aim of this study is to confirm the presence and molecular identification of Echinococcus tapeworms in wolves from south-eastern Poland. An investigation was carried out on the intestines of 13 wolves from south-eastern Poland. The small intestines were divided into three equal segments. Each segment was separately examined using the sedimentation and counting technique (SCT). The detected Echinococcus tapeworms were isolated and identified by PCRs and sequencing (nad1 and cox1 genes). Additionally, DNA isolated from the feces of wolves positive for Echinococcus tapeworms was examined with two diagnostic PCRs. The intestines of one wolf were positive for E. granulosus s.l. when assessed by SCT; the intestine was from a six-year-old male wolf killed in a communication accident. We detected 61 adult tapeworms: 42 in the anterior, 14 in the middle, and 5 in the posterior parts of the small intestine. The PCRs conducted for cox1 and nad1 produced specific products. A sequence comparison with the GenBank database showed similarity to the deposited E. ortleppi (G5) sequences. An analysis of the available phylogenetic sequences showed very little variation within the species of E. ortleppi (G5), and identity ranged from 99.10% to 100.00% in the case of cox1 and from 99.04% to 100.00% in the case of nad1. One of the two diagnostic PCRs used and performed on the feces of Echinococcus-positive animals showed product specific for E. granulosus. This study showed the presence of adult E. ortleppi tapeworms in wolves for the first time.

Distribution of Parasitic Helminths in the Small Intestine of the Red Fox (Vulpes vulpes).

The aim of the study was to analyze the distribution of the main groups of parasitic helminths within the small intestine of the red fox on the example of animals coming from eastern Poland. Two hundred and sixteen red foxes shot in eastern Poland were used in the investigation. Before examination, each small intestine was divided into three equal parts: anterior (A), middle (M), and posterior (P). Each part was examined separately with the sedimentation and counting technique. Six different types of intestinal parasites were detected: Alaria alata (78.7%), Mesocestoides spp. (78.2%), hookworms (72.7%), Taenia spp. (53.2%), Toxocara/Toxascaris (43.1%), and Echinococcus multilocularis (18.5%). Alaria alata was most often found in A and in the only-A variant. Taenia spp. and Toxocara/Toxascaris occurred often in A and were the second (after A. alata) parasites in terms of frequency occurring in the only-A variant. Mesocestoides spp. was most commonly located in M. Parasites with predilection sites located mainly in M and P were E. multilocularis and hookworms. In all parasite species, the variant covering the entire intestine (A + M + P) was found in samples with a higher intensity compared to variants limited to one or two fragments. Our investigation, as one of the few of its type, conducted a comprehensive analysis of the distribution of intestinal helminths in the small intestine of the red fox. It showed significant differences in the distribution of parasitic helminths in the small intestine of the red fox. Determining typical predilection sites for parasites in the intestine can be helpful in creating effective diagnostic methods.

Tritrichomonas Foetus: A Study of Prevalence in Animal Hosts in Poland.

Tritrichomonas foetus is described as a pathogen of cattle and cats and also exhibits commensalism with pigs. In order to estimate the prevalence and determine the risk factors for parasite infection, specimens from animal hosts (cat, pigs, and cattle) from Poland were investigated. To our best knowledge, this is the first such study to examine samples from wild boars (Sus scrofa) for the presence of T. foetus. Data were collected from 117 cats, 172 pigs, 236 wild boars, and 180 cattle. The sensitivity of T. foetus identification was increased by using two molecular assays: PCR and loop-mediated isothermal amplification (LAMP). The prevalence of feline tritrichomonosis was 20.51%, and statistically significant differences were obtained between groups of animals regarding age, breed, number of cats, diarrhea, and place of living. Positive PCR and LAMP results for T. foetus were estimated for 16.28% of pigs, and the obtained data were significantly correlated with age. Conversely, no significant differences were observed concerning the farm size factor. In our survey, no cases of bovine tritrichomonosis were found, which is consistent with the data from the other countries of the European Union. Similarly, all wild boar samples were also T. foetus-negative according to LAMP and PCR.

Diversity of Trichinella species in relation to the host species and geographical location.

Trichinella nematodes still circulate in various hosts in both domestic and sylvatic environments. Recently, in Europe, the transmission of Trichinella spp. to humans has been attributed more to wild animals than to domestic animals. However, domestic animals could still be a source of human infections in some regions. Therefore, our aim was to determine the species composition of Trichinella and the prevalence and intensity of infections in animal populations from the domestic cycle, namely pigs (Sus scrofa f. domestica); the synantropic cycle, in the form of rats (Rattus norvegicus); and the sylvatic cycle, namely wild boars (Sus scrofa) and red foxes (Vulpes vulpes), in Poland. The findings showed that the nematode prevalence in pigs (0.0002 %) and wild boars (0.3 %) was lower than it was in red foxes (4 %). A very high prevalence was found in rats (23.3 %), but it must be emphasized that the investigated rat samples were collected from farms where pigs were infected with Trichinella spp. The mean larval burden was found to be higher in wild boars and pigs (11.48 lpg and 10.19 lpg) than in red foxes and rats (4.09 and 2.30). Trichinella spiralis was the predominant species in pigs (98.6 %), wild boars (77.3 %) and rats (100 %), while in red foxes, this species occurred less frequently (15.5 %). The most frequently occurring species in red foxes was Trichinella britovi (73.2 %). Moreover, in wild boar and red fox coinfections, T. spiralis/T. britovi were detected (3.1 and 9.9 %, respectively). In addition, Trichinella pseudospiralis was detected in a few wild boars (0.5 %) and Trichinella nativa was found in one red fox and one wild boar. Furthermore, different T. spiralis and T. britovi prevalence ratios in various geographical regions were found. In the wild boar population, a higher frequency of T. spiralis (70-85 % of infected animals) was observed in the western and central parts of Poland, while in the eastern part, this dominance was not as evident (46-59 %). In the red fox population, T. britovi was abundant throughout the entire territory; however, its highest prevalence was in the east (90-100 %).

First case of Trichinella spiralis infection in beavers (Castor fiber) in Poland and Europe.

BACKGROUND: This is the first report of the finding of Trichinella spiralis in beaver meat (Castor fiber) in Poland and Europe. In Poland, the beaver is a strictly protected animal species, except the few regions where high population density leads to economic losses. In these areas, the reduction culling of the animals was introduced. This uncommon hunting game animal is consumed and treated as a delicacy by hunters. However, currently, there is a lack of knowledge on possible risk factors for humans associated with the consumption of beaver meat. This paper presents the result of the study on the occurrence of nematodes of the genus Trichinella in beavers. METHODS: In total, 69 beavers were examined for the presence of Trichinella spp. The 50g samples were taken from each animal and digested separately, according to a procedure based on the EU reference method. The larva DNA was examined by PCR and sequencing. RESULTS: One of the 69 examined beavers was infected. Only one Trichinella larva was detected by the digestion method. The result of PCR confirms the presence of T. spiralis in beaver meat. CONCLUSIONS: This case further confirms the ability of these typical herbivores to be infected with Trichinella spp. This is the second confirmed case of Trichinella spp. infection in beavers in Europe and the first of T. spiralis.

Intraspecific genetic variation in Trichinella spiralis and Trichinella britovi populations circulating in different geographical regions of Poland.

Trichinella spiralis and Trichinella britovi are species of nematodes which are responsible for the majority of Trichinella infections in the world and the most prevalent in Poland. The most abundant species - T. spiralis, is considered to be more genetically homogeneous in Europe than T. britovi. The aim of the present study was to determine the genetic variability in T. spiralis and T. britovi populations based on nuclear 5S rDNA intergenic spacer region (5S rDNA) and cytochrome c oxidase 1 (COX1) gene sequences. For the study, 55 isolates of T. spiralis and 50 isolates of T. britovi isolated from wild boars, pigs, brown rat and a red fox were analyzed. Based on the analysis of both genes, the genetic variability within populations of T. spiralis and T. britovi differed. In T. spiralis, two single nucleotide polymorphisms (SNPs) were observed in the 612 bp 5S rDNA gene fragment, and one SNP was detected in the 700 bp COX1 gene fragment. In T. britovi, 17 single nucleotide variations (SNVs) were detected in the 5S rDNA gene fragment (among them 16 SNPs), while COX1 sequence analysis revealed the occurrence of 20 SNVs between the sequences tested (among them 19 SNPs). For the majority of T. spiralis isolates the investigated larvae presented uniform haplotypes. In contrast, most of the isolates of T. britovi consisted of larvae of different haplotypes. Geographical analysis showed that each region exhibited different haplotype composition and richness. Warminsko-Mazurskie and Zachodniopomorskie regions were the richest in haplotypes (15 and 16 haplotypes, respectively). We used heatmaps showing a characteristic pattern for each region graphically. This may allow to differentiate regions based on the occurrence of particular haplotypes. Furthermore, a PCA analysis on the SNP level yielded biplots that show that certain haplotypes/genotypes are associated with (clusters of) regions.

First report of Echinococcus multilocularis in cats in Poland: a monitoring study in cats and dogs from a rural area and animal shelter in a highly endemic region.

BACKGROUND: Alveolar echinococcosis is a dangerous zoonotic disease caused by larval forms of Echinococcus multilocularis. In its life-cycle, the principal definitive host is the red fox; however, domesticated carnivorous animals (dogs and cats) can also act as definitive hosts. Until now, there were no data concerning this infection in cats in Poland. The aim of this study was to estimate the prevalence of E. multilocularis in cats and dogs originating from rural areas and animal shelters in a region characterised by a high prevalence of E. multilocularis in red foxes. METHODS: Samples of faeces were collected from 67 cats and 268 dogs from a rural area (villages and animal shelters) of a highly endemic region in southeastern Poland. Samples were examined using nested PCR (E. multilocularis), multiplex PCR (E. multilocularis, Taenia spp.) and PCR [E. granulosus (s.l.)]. Additionally, faeces were examined microscopically (flotation). Moreover, intestines from 110 red foxes shot in the investigated area were examined (sedimentation and counting technique). RESULTS: Positive PCR results for E. multilocularis were obtained in 4 cats (6.0%) and 4 dogs (1.5%). There were no significant differences between groups of animals (from a shelter and with an owner) concerning the prevalence of E. multilocularis in both cats and dogs. Taenia spp. were found in 10 cats (14.9%) (Taenia taeniaeformis and T. hydatigena) and 26 dogs (9.7%) (T. hydatigena, T. serialis, T. taeniaeformis, T. crassiceps, T. pisiformis and T. ovis) and Mesocestoides litteratus was found in 4 cats (6.0%) and 3 dogs (1.1%). All samples were negative for E. granulosus by PCR. Taking into consideration PCR and flotation results, 29 cats (43.3%) and 73 dogs (27.2%) were infected with helminths (26.9 and 11.9%, respectively, were infected with tapeworms). The highly endemic status of the investigated area was confirmed by examination of red foxes: 48.2% of examined red foxes were infected with E. multilocularis. CONCLUSIONS: To the best of our knowledge, this study reports the presence of E. multilocularis in cats for the first time in Poland and confirms the role of dogs in this infection in highly endemic areas.

Parasitological contamination with eggs Ascaris spp., Trichuris spp. and Toxocara spp. of dehydrated municipal sewage sludge in Poland.

The objective of the present study was to evaluate the contamination of sewage sludge produced by municipal waste treatment plants in Poland by viable eggs of intestinal parasites of the genera Ascaris, Toxocara and Trichuris (ATT). Ninety-two municipal, mechanical-biological sewage treatment plants located within Poland were selected. These plants belonged to types of agglomerations: group 0 (large), group 1 (medium), group 2 (smaller) and group 3 (small). Samples were collected at the final stage of sewage treatment after the addition of flocculent to sludge, followed by dehydration. The samples were examined by a method adjusted to examine sewage sludge dehydrated using polyelectrolytes. The viability of the isolated eggs was evaluated based on incubation in a moist chamber. Live eggs of intestinal nematodes were found in 99% of samples. Most samples were contaminated by the eggs of Ascaris spp. (95%) and Toxocara spp. (96%). However, Trichuris spp. eggs were detected in 60% of samples. The mean number of eggs in 1 kg of dry mass (eggs/kg d.m.) was 5600 for Ascaris, 3700 for Toxocara and 1100 for Trichuris. The highest number of ATT eggs was detected in samples from sewage treatment plants located in south-eastern and central Poland. The highest number of ATT eggs was found in sewage sludge produced in large sewage treatment plants (agglomeration Groups 0 and 1), with mean values of 15,000 and 8900 eggs/kg d.m. The present study is the first parasitological investigation conducted on a large number of samples (92 samples) taken from various types of municipal sewage treatment plants located throughout Poland (16 regions) after the common introduction of polyelectrolytes during sewage sludge dehydration. The results of this study indicate that sludge produced in municipal sewage treatment plants is highly contaminated with parasite eggs.

Tritrichomonas Foetus as a Causative Agent of Tritrichomonosis in Different Animal Hosts.

Tritrichomonas foetus is a protozoan parasite that has been traditionally identified as a cause of reproductive tract disease in cattle and gastrointestinal tract infection in cats. Moreover, T. foetus is also well known as a commensal of the nasal cavity, intestines, and stomach in swine. In this review we describe T. foetus as a pathogen dangerous to more than one animal host, diagnostic and taxonomic aspects of this infection, and the extent to which isolates from different hosts share genetic identity.

Negative effect of flocculant (cationic acrylamide) on detectability of the nematode eggs in sewage sludge.

The use of sewage sludge in agriculture brings the risk of microbiological and parasitological contamination of soil, ground and surface water, as well as cultivated plants. Therefore prior the application to the soil, sewage sludge must be examined, among others, for the presence of live eggs of intestinal parasites. However, the efficiency of commonly used for this purpose parasitological methods is not satisfactory. This is probably due to the presence of flocculants in the sediments used in the dehydration process. The objective of the study was analysis of the effect of flocculant (cationic acrylamide) on the possibilities of isolation of parasite eggs from dehydrated sewage sludge. For this purpose 10 samples of sewage sludge were prepared: 5 containing flocculant and 5 without flocculant. Samples were tested by flotation method according to Quinn. From sewage sludge free of flocculant, 67.8 eggs were isolated, on average, whereas from sludge containing flocculant - only 2.8 eggs. The experiments confirmed that the isolation of eggs from sewage sludge containing flocculant (cationic acrylamide) is much more difficult than from sludge free from this substance and therefore the simple parasitological methods should not be used to examine the dehydrated sewage sludge.

Occurrence of Trichinella spp. in rats on pig farms.

INTRODUCTION: The highest risk of trichinellosis for human is considered in eating meat products containing live larvae, mostly from wild boars or pigs. Spreading of Trichinella spp. may occur in various ways, one of which is transmission by vectors. The rat is considered to be the most common vector for Trichinella parasite. The population of rats living on pig farms can play an important role in maintaining or spreading the parasite to other animals. OBJECTIVE: The aim of presented survey was to investigate the occurrence of Trichinella spp. in rats on farms with pigs infected with this parasite. MATERIAL AND METHODS: From pig farms selected for study, the muscles of collected rats were investigated by magnetic stirrer digestion method to assess occurrence of Trichinella in the rat population. Isolated Trichinella parasites were identified under stereomicroscope and multiplex PCR were performed for species identification. RESULTS: Rats infected with Trichinella spp. were discovered on three of five investigated pig farms. The mean extent of invasion in rats from the studied farms was 23.33%. The calculated medium intensity of invasion was 4.09 lpg (larvae per gram) (SD 5.41). All larvae of Trichinella discovered from rats were identified as T.spiralis. CONCLUSIONS: The results obtained indicate that in farms with a high prevalence of Trichinella invasion in pigs there are very likely to be found rats infected by this nematode. This suggests possibility to maintain the invasion in herd and spread into neighborhood farms.

Prevalence of intestinal helminths of red foxes (Vulpes vulpes) in central Europe (Poland): a significant zoonotic threat.

BACKGROUND: The red fox (Vulpes vulpes) is widely distributed in the world; in central Europe, it is the most numerous wild species of the family Canidae. It can play the role of a definitive host for many intestinal parasites, including zoonotic helminths. Poland, with its geographical location (central Europe), is an interesting area for parasitological investigations of this species. The aim of this study was to evaluate and compare the prevalence of intestinal helminths in red foxes in different regions of Poland. METHODS: Intestines of 473 red foxes from four different regions were examined using the sedimentation and counting technique (SCT). In addition, 344 samples of faeces were examined using flotation. RESULTS: Overall, intestinal helminths were found in 98.9% of red foxes. The average prevalence of detected parasites was as follows: Mesocestoides spp. (84.1%); hookworms (67.9%); Alaria alata (61.5%); Toxocara/Toxascaris (49.5%); Taenia spp. (42.5%); Echinococcus multilocularis (25.6%); and Trichuris vulpis (2.3%). The prevalence of the majority of parasite species was similar in each region. Significant differences between regions were observed in the case of E. multilocularis: a low prevalence in the south-western and northern regions (0% and 0.9%, respectively) and a high prevalence in the south-east and northeast (39.3% and 42.7%, respectively). In the case of A. alata, important differences were found between northern (96.5% and 93.7% in northern and northeast regions, respectively) and southern regions (15.2% and 24.7% for south-western and south-east regions, respectively). The percentage of positive samples obtained with coproscopic examination (except for Trichuris) was significantly lower than that obtained with SCT. Analysis of the prevalence estimated in individual regions with the use of both methods (flotation and SCT) showed a high correlation for all parasite species (except for Mesocestoides spp.). The flotation method also allowed us to detect the eggs of the lung nematode Eucoleus aerophilus (syn. Capillaria aerophila) (76.2% of positive foxes). CONCLUSIONS: This study showed a very high percentage of red foxes infected with intestinal helminths in different parts of Poland. Depending on the location, some differences were observed regarding the prevalence of dangerous zoonotic parasites, which should be considered in the assessment of infection risk for humans.