Isoelectric Focusing of Proteins in The pH Gradient As a Tool for Identification of Species Origin of Raw Meat.

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.

First case of Trichinella nativa infection in wild boar in Central Europe-molecular characterization of the parasite.

The examination of wild boars gained in Poland shows for the first time occurrence of Trichinella nativa, freeze-resistant species of Trichinella in this host from the central Europe region. This finding is not only one of several cases of T. nativa invasion in wild boars all over the world but also one of the very few cases of T. nativa detected so far beyond the known boundary of occurrence of this species. The molecular characterization of discovered larvae based on analysis of partial genes: 5s rDNA-ISR and CO1 confirm the findings. Moreover, the analyzed DNA sequences of both genes present new haplotypes of T. nativa in comparison to that described previously.

Genetic diversity of Echinococcus multilocularis in red foxes in Poland: the first report of a haplotype of probable Asian origin.

The aim of the present study was to estimate the genetic diversity of the cestode Echinococcus multilocularis Leuckart, 1863 in Poland based on sequence analysis of the mitochondrial genes of worms isolated from red foxes, Vulpes vulpes (Linnaeus). Overall, 83 adults of E. multilocularis from the same number of foxes in different parts of Poland were used for analysis. Sequences of the three mitochondrial genes, cytochrome b (cob), NADH dehydrogenase subunit 2 (nad2) and cytochrome c oxidase subunit 1 (cox1), were analysed. Seventy-four individual biological samples were successfully sequenced. Combined sequence analysis of these three genes exhibited fifteen Polish haplotypes (EmPL1-EmPL15). Most isolates (n = 29; 39%) were classified to the EmPL1 haplotype, which occurred mainly in the east, north and centre of Poland. Haplotype EmPL4 (n = 14; 19%) and other haplotypes appeared predominantly in the south and west area. Fourteen haplotypes were grouped in the European clade. One Polish haplotype (EmPL9) (n = 7, 10%) was assigned to the Asian clade with haplotypes from Japan and Kazakhstan. This haplotype was found only in northeast Poland and this is the westernmost report of haplotype of E. multilocularis belonging to the Asian clade in Europe. The investigation demonstrated that populations of E. multilocularis in Poland (and probably also in eastern Europe) included not only different European haplotypes but also those of the Asian origin.

The prevalence of Echinococcus multilocularis in red foxes in Poland--current results (2009-2013).

The aim of the study was to determine the prevalence of Echinococcus multilocularis in red foxes (Vulpes vulpes) in Poland. Overall, 1,546 intestinal samples from 15 of the 16 provinces in Poland were examined by the sedimentation and counting technique (SCT). The mean prevalence of E. multilocularis in Poland was 16.5% and was found in 14 of the 15 examined provinces. The mean intensity of infection was 2,807 tapeworms per intestine. Distinct differences in prevalence were observed between regions. In some provinces of eastern and southern Poland, the level of prevalence was 50.0% (Warminsko-Mazurskie), 47.2% (Podkarpackie), 30.4% (Podlaskie) and 28.6% (Malopolskie), while in other provinces (west and south-west), only a few percent was found: 2.0% (Dolnoslaskie), 2.5% (Wielkopolskie) and 0.0% (in Opolskie). The border between areas with higher and lower prevalence seems to coincide with a north-south line running through the middle of Poland, with prevalence from 17.5 to 50.0% in the eastern half and from 0.0 to 11.8% in the western half. The dynamic situation observed in the prevalence of this tapeworm indicated the necessity of continuing to monitor the situation concerning E. multilocularis in red foxes in Poland.

Trichinella species circulating in wild boar (Sus scrofa) populations in Poland.

Hunting in Poland has a long tradition and became more popular after 1990. Each year over 60,000 wild boar are hunted. Some of them may act as Trichinella carriers thus all carcasses of wild boar are systematically sampled in game-handling establishments as part of the post-mortem examination. The aim of the study was to determine the species of Trichinella and to evaluate the year to year differences in the occurrence of those species in the populations of wild boar in Poland. Samples for the study were provided by the Veterinary Inspection Service. Wild boar carcasses were examined using a digestion method. Only samples recognized as positive for Trichinella in these examinations were sent to the National Reference Laboratory (NRL) for confirmation of genus identity. Samples from 450 animals were obtained for the study (380 muscle samples and 70 larval isolates preserved in 90% ethyl alcohol). Tissue samples were digested to isolate larvae. Extracted larval DNA was amplified using a modified multiplex PCR protocol to identify the species of Trichinella. Five larvae from each sample were examined by PCR. The study revealed that Trichinella spiralis and Trichinella britovi are present in wild boar in Poland in a ratio of 3:1. Mixed infections with T. spiralis and T. britovi were found in 1% of the animals.