Validation process of automatic DNA extraction from bone material using a new advanced protocol for the EZ2 Connect instrument.

Identification of human remains using genetic methods is an important task of forensic science. DNA markers are proving essential in the identification of unknown human remains. However, environmental factors can lead to poor preservation of DNA, including in bone material. The aim of this study was therefore to compare two methods of DNA isolation from bone material: the traditional organic method and the new protocol using the EZ2 Connect instrument. The study involved three types of bone material, namely molars/premolars, petrous parts of the temporal bone and femurs, all with an estimated PMI of 70-80 years. Importantly, the biological material was obtained from three different environments, categorized as preserving, neutral and degrading, based on basic physico-chemical tests and the potential impact on the bone. The results obtained show that the DNA was best preserved in the petrous bone, followed by the teeth, and the femur. DNA extraction using the EZ2 Connect instrument with a new protocol gave slightly better results for the petrous bone, comparable results for the teeth and worse results for the femur compared to the organic method. Several protocol modifications were tested and optimal conditions for DNA isolation were proposed for the EZ2 protocol. Furthermore, the use of an automated method facilitated the effective accumulation of isolates and increased the chances of successful identification of unknown human remains.

Genetic differences in variants of the AMD virus at the site of a disease outbreak.

Aleutian Mink Disease Virus (AMDV) results in mink breeding losses due to frequent abortion, low fecundity and high juvenile mortality. Due to the high persistence of pathogen in the environment and lack of causative treatment there is a need for research on alternative methods to eliminate the pathways of the spread of the virus and extinguish current outbreaks. The aim of the study was to investigate molecular variation of AMDV on a farm where mass deaths of mink took place. The material for the research was obtained from a mink farm located in Latvia. Mass deaths had occurred on the farm among symptoms typical of Aleutian disease. Spleen samples were collected from the dead animals during post-mortem examination. Sequencing and bioinformatic analysis made it possible to distinguish the variants occurring in the groups. The presence of the genetic material of the virus was confirmed by PCR and qPCR in each of the spleen samples. The isolates were divided into two main groups: the dominant group A, with more than 83% of all isolates, and group B. Comparison of the variants with the nonpathogenic strain AMDV-G revealed that isolates from group A were more than 95% similar to that strain, whereas the similarity of group B isolates was just over 86%. The average viral load in both groups was 108 copies; no differences in viral load were noted between groups. Testing based on serological analysis produces fairly effective screening results, but these methods do not enable complete elimination of the virus from a population. Only their use in combination with modern testing techniques as tools for identification of vectors and the directions of the spread of the AMD virus can make it possible to block the routes of its spread and to extinguish its current outbreaks.

Meat Nutritional Value of Pulawska Fattening Pigs, Polish Large White × Pulawska Crossbreeds and Hybrids of DanBred.

The nutritional quality of meat and the health-promoting properties of intramuscular fat from randomly selected rearing pigs of the native Pulawska breed (PUL, n = 15) and its crossbreeds with the Polish Large White breed (PLW × PUL, n = 16) and the DanBred hybrids (DAN, n = 17) were compared. The typical commercial fattening period was carried out up to 80 days of age. The initial body weight of PUL weaners was 30.55 kg (±3.98 kg), that of PLW × PUL weaners was 30.00 kg (±4.29 kg) and that of DAN weaners was 30.70 kg (±3.40 kg). The protein content and energy value of the complete feeds (Grower and Finisher) were 165 and 155 g/kg, and 12 and 11.8 MJ/kg, respectively. The research material consisted of samples of Longissimus lumborum and Semimembranosus. The chemical analyses included the determination of proximate composition, fatty acid content and amino acid concentration. The lowest fat content was found in the meat of the Pulawska pigs, while the highest was found in the DanBred fattening pigs. The highest content of SFAs and MUFAs was found in the meat of DanBred hybrids, while it was significantly lower in Pulawska pigs. The genetic group did not affect the content of PUFAs, including n-3 and n-6. Significantly lower and more favourable atherogenic and thrombogenic indices were found for the intramuscular fat of Pulawska pigs. The very limited effect of the genetic group on amino acid content was observed, except for that of serine, histidine and lysine. In general, the meat of pigs of the Pulawska breed showed the highest nutritional value and the most favourable health-promoting properties.

The Problem of Antimalarial-Drug Abuse by the Inhabitants of Ghana.

Introduction: Malaria is still a huge social and economic health problem in the world. It especially affects the developing countries of Africa. A particular problem is the misuse and abuse of over-the-counter antimalarials. This problem could lead to the emergence of drug-resistant strains and the subsequent elimination of more antimalarials from the list of effective antimalarials in Ghana. Methods: During the implementation of the study, an original questionnaire was used to collect data among Ghanaians on their knowledge of malaria, attitude towards antimalarials and their use of antimalarials. Results: The proportion in the analyzed subgroups was compared using the chi-square test. The analysis was conducted using TIBCO Software Inc., Krakow, Poland (2017) and Statistica (data analysis software system), version 13. In total, 86.29% of respondents knew the symptoms of malaria (p = 0.02) and 57.2% knew the cause of malaria (p < 0.001). Respondents with higher education were significantly more likely to know the symptoms of malaria (96%) p < 0.001. In the study group, only 74.59% of the respondents consulted medical personnel before taking the antimalarial drug (p = 0.51) and only 14.2% of the remaining respondents performed a rapid diagnostic test for malaria. Conclusions: The awareness of Accra and Yendi native inhabitants about the causes and symptoms of malaria and alternative ways of prevention is quite high. People's education very significantly influences the way Accra residents deal with suspected malaria. Widespread public education and awareness and accessibility to places where antimalarial drugs are sold play a very important role in the proper use of antimalarial drugs.

Effect of the addition of chokeberry leaf extract on the physicochemical and sensory properties of burgers from dark cutting veal.

The occurrence of the DFD defect shortens microbiological stability and reduces consumer acceptance of meat. The effect of a chokeberry leaf extract (ChLE) addition at concentrations of 0.01 %, 0.05 % and 0.1 % on the sensory and physicochemical properties of burgers prepared from DFD meat during refrigerated storage under vacuum for 10 days was assessed. The pH, water activity, texture, colour, degree of lipid oxidation, and content of α-tocopherol were measured. The extract at concentrations of 0.05 % and 0.1 % significantly influenced lower levels of lipid oxidation and higher content of α-tocopherol on each tested period. Moreover, these concentrations had a beneficial effect on the instrumental and sensory evaluation of texture parameters and on overall quality. It was concluded that the addition of 0.1 % extract was the most beneficial over the entire 10-day storage period. In summary, use of ChLE may reduce the loss of raw meat caused by DFD defect.

The Effect of Low Doses of Acetylsalicylic Acid on the Occurrence of Rectal Aberrant Crypt Foci.

Background and Objectives: Aberrant crypt foci (ACF) are one of the earliest putative preneoplastic and, in some cases, neoplastic lesions in human colons. Many studies have confirmed the reduction of ACFs and colorectal adenomas after treatment with acetylsalicylic acid (ASA) commonly referred to as ASA; however, the minimum effective dose of ASA and the duration of use has not been fully elucidated. The objective of our study was to assess the significance of low dose ASA (75-mg internally once daily) to study the chemopreventive effect of ASA in ACF and adenomas development in patients taking this drug for a minimum period of 10 years. Materials and Methods: Colonoscopy, combined with rectal mucosa staining with 0.25% methylene blue, was performed on 131 patients. The number of rectal ACF in the colon was divided into three groups: ACF < 5; ACF 5−10; and ACF > 10. Patients were divided into two groups: the "With ASA" group (the study group subjects taking ASA 75-mg daily for 10 years); and "Without ASA" group (control group subjects not taking ASA chronically). The incidence of different types of rectal ACF and colorectal polyps in both groups of subjects was analysed and ascertained. Results: Normal ACF was found in 12.3% in the study group vs. 87.7% control group, hyperplastic 22.4% vs. 77.6%, dysplastic 25% vs. 75%, mixed 0% vs. 100%. Treatment with ASA affects the occurrence of colorectal adenomas. The amount of dysplastic ACFs was lower in the study group than in the control group. The increase in dysplastic ACFs decreases with age in both groups, with the increase greater in those not taking ASA. Conclusions: Patients who take persistent, chronic (>10 years) low doses of ASA have a lower total number of all types of rectal ACFs and adenomas compared to the control group.

Genotypes and Variants of BKPyV in Organ Donors after Brain Death.

Kidney transplantation from a donor with latent BKPyV might be the cause of serious complications, such as BK virus-associated nephropathy. The aim of the study was to determine the prevalence of BKPyV infection in donors after brain death (DBDs), to analyse the molecular variation of BKPyV and to compare clinical and inflammation parameters of DBDs infected with various genotypes of BKPyV. BKPyV was investigated in blood and urine samples of 103 DBDs using PCR followed by sequencing and bioinformatic analysis, and the viral load was assessed by qPCR. Clinical parameters, including cellular markers of inflammation were assessed. The results confirm high prevalence of BKPyV (48%),and genotype IV (49%) over genotype I (43%) and the co-infection with genotypes I and IV in 8.2%. Viral load ranged from 102 to 107 copies/mL, with an average of 1.92 × 106 copies/mL. No specific markers for BKPyV infection were detected among the parameters tested. Infection with genotype I may be associated with the adverse impact on thekidney function, while infection with genotype IV was associated with the anemia Not only the viral load but also the genotype of BKPyV may have an impact on the course of infection.

Effect of heat treatments on the physicochemical and sensory properties of the longissimus thoracis muscle in unweaned Limousin calves.

The aim of the study was to assess the effect of methods of heat treatment on selected quality parameters of longissimus thoracis muscle of Limousin calves, subjected to grilling, steaming, and sous vide cooking. The type of heat treatment did not significantly affect shear force or water activity. Cooking loss in the grilled and steamed meat was significantly higher than in the sous vide. While the content of vitamin E was significantly lower in the cooked meat, it was retained to the greatest degree in the grilled meat and least in the steamed meat. The TBARS index significantly increased during all cooking methods with the highest level in steamed meat. The sensory analysis revealed a preference for the sous vide and grilled meat, while the steamed meat received the lowest scores. Although veal is commonly assumed to be highly nutritious and palatable, its quality can be significantly influenced by the type of heat treatment.

Molecular Epidemiology and Variation of the BK Polyomavirus in the Population of Central and Eastern Europe Based on the Example of Poland.

The BK polyomavirus (BKPyV) is a widespread pathogen in humans. Polymorphism of the region encoding the VP1 protein of BKPyV provides the basis for classifying the virus into types and subtypes, whose frequency varies depending on geographic location. The aim of our study was to determine the frequency of BKPyV in the Polish population and to assess its variation by analysing polymorphism in the typing region. The study was conducted on 168 healthy, Polish volunteers, whose blood (plasma) and urine were sampled. The virus was detected using PCR, products, sequenced and subjected to bioinformatic analysis. In addition, viral load was assessed by qPCR. The presence of the genetic material of the BK virus was noted in 61/168 urine samples but in none of the plasma sample. Sequencing and phylogenetic analysis confirmed that the BKPyV isolates were of types I and IV, dominant in Europe (63.93% and 36.07%, respectively). All isolates from genotype I belonged to subtype Ib-2, showing polymorphism at position 1809 with a frequency of 61.54% (G1809A) and 38.46% (G1809C). To the best of our knowledge, this is the first study of this magnitude on the genetic variation of BKPyV among healthy volunteers in Poland.

BK Polyomavirus-Biology, Genomic Variation and Diagnosis.

The BK polyomavirus (BKPyV), a representative of the family Polyomaviridae, is widespread in the human population. While the virus does not cause significant clinical symptoms in immunocompetent individuals, it is activated in cases of immune deficiency, both pharmacological and pathological. Infection with the BKPyV is of particular importance in recipients of kidney transplants or HSC transplantation, in which it can lead to the loss of the transplanted kidney or to haemorrhagic cystitis, respectively. Four main genotypes of the virus are distinguished on the basis of molecular differentiation. The most common genotype worldwide is genotype I, with a frequency of about 80%, followed by genotype IV (about 15%), while genotypes II and III are isolated only sporadically. The distribution of the molecular variants of the virus is associated with the region of origin. BKPyV subtype Ia is most common in Africa, Ib-1 in Southeast Asia, and Ib-2 in Europe, while Ic is the most common variant in Northeast Asia. The development of molecular methods has enabled significant improvement not only in BKPyV diagnostics, but in monitoring the effectiveness of treatment as well. Amplification of viral DNA from urine by PCR (Polymerase Chain Reaction) and qPCR Quantitative Polymerase Chain Reaction) is a non-invasive method that can be used to confirm the presence of the genetic material of the virus and to determine the viral load. Sequencing techniques together with bioinformatics tools and databases can be used to determine variants of the virus, analyse their circulation in populations, identify relationships between them, and investigate the directions of evolution of the virus.

Detection of mink astrovirus in Poland and further phylogenetic comparison with other European and Canadian astroviruses.

Mink astrovirus infection remains a poorly understood disease entity, and the aetiological agent itself causes disease with a heterogeneous course, including gastrointestinal and neurological symptoms. This paper presents cases of astrovirus infection in mink from continental Europe. RNA was isolated from the brains and intestines of animals showing symptoms typical of shaking mink syndrome (n = 6). RT-PCR was used to amplify astrovirus genetic material, and the reaction products were separated on a 1% agarose gel. The specificity of the reaction was confirmed by sequencing fragment coding RdRP protein (length of sequencing product 170 bp) from all samples. The presence of astrovirus RNA was detected in each of the samples tested. Sequencing and bioinformatic analysis indicated the presence of the same variant of the virus in all samples. Comparison of the variant with the sequences available in bioinformatics databases confirmed that the Polish isolates form a separate clade, closely related to Danish isolates. The dissimilarity of the Polish variant to those isolated in other countries ranged from 2.4% (in relation to Danish isolates) to 7.1% (in relation to Canadian isolates). Phylogenetic relationships between variants appear to be associated with the geographic distances between them. To our knowledge, this work describes the first results on the molecular epidemiology of MAstV in continental Europe. The detection of MAstV in Central Europe indicates the need for further research to broaden our understanding of the molecular epidemiology of MAstV in Europe.

Effectiveness of various methods of DNA isolation from bones and teeth of animals exposed to high temperature.

In the event of fires, natural disasters, and other events associated with high temperature, bones and teeth are the only source of genetic material for identifying human or animal carcasses. To obtain reliable final results of identification tests, the use of appropriate nucleic acid extraction methods is crucial. Therefore, the main objective of this research was to evaluate the effectiveness of selected methods of DNA isolation from animal burnt bones and teeth. In addition, the effect of the duration of high temperature on the stability of nuclear and mitochondrial DNA in these tissues was determined, as well as the possibility of using the genetic material obtained for species identification of remains of unknown origin. Bones and teeth collected during necropsy of dogs were burnt in a laboratory oven at 400 °C (752 °F; 673.15 K) for 5, 10, 15, 30, 45 and 60 min. DNA was isolated according to four different protocols, using three commercial kits, i.e. the PrepFiler® Forensic DNA Extraction Kit from Applied Biosystems, the QIAamp® DNA Investigator Kit from QIAGEN, and the DNA Mini Kit from Syngen, as well as a classic organic method. The effectiveness of these methods was compared by assessing the amount of isolated DNA using Real-Time PCR and its purity using a NanoDrop™ spectrophotometer. Each isolate was also subjected to PCR with primers designed to amplify fragments of dog mitochondrial DNA. The effectiveness of species identification was assessed for the method showing the best DNA recovery and for the organic method, considered the gold standard for analysis of difficult material. The QIAamp® DNA Investigator Kit showed the highest efficiency of DNA isolation from bones and teeth burnt for 15 min (the longest burning time for which DNA could still be recovered from bones and teeth). The results of the experiment clearly indicate that DNA stability in hard tissues depends on how long they burn. In the case of exposure to 400 °C, reliable genetic testing, including species identification, is possible when the burning time does not exceed 15 min. Among the hard tissues examined, bones proved more suitable than teeth for identification purposes. It was also concluded that identification of bone remains with extreme heat damage should be based on mitochondrial DNA analysis.

The Effect of Smoking on the Number and Type of Rectal Aberrant Crypt Foci (ACF)-First Identifiable Precursors of Colorectal Cancer (CRC).

BACKGROUND: The problem of smoking and its influence on the occurrence of precursors and advanced colorectal cancer is often discussed in the medical literature. Tobacco smoke can provide a non-nutritional source of polycyclic hydrocarbons and other substances which, when combined with an incorrect diet, may play a role in promoting carcinogenesis at the level of the genetic control mechanism. The aim of our study was to assess the effect of smoking on the frequency and type of aberrant crypt foci (ACF) in the rectum and polyps in the large intestine in people who smoke more than 20 cigarettes a day for more than 20 years. METHODS AND FINDINGS: A colonoscopy combined with rectal mucosa staining with 0.25% methylene blue was performed in 131 patients. Each of the study participants gave informed consent to participate in the study. Three bioptates were collected from the foci defined macroscopically as ACF; in cases where there were fewer foci, the number of collected foci was respectively lower. On the colonoscopy day, patients completed the questionnaire regarding epidemiological data used for analysis of factors affecting the occurrence of ACF in the study group. The number of ACF in the colon was divided into three groups: -ACF < 5, 5 < ACF < 10, ACF > 10. In the statistical analysis, numerical data were presented and real numbers, range of arrhythmic means, mean standard deviation, and results of probability distribution. The Student's test, U test, and chi2 were applied in order to determine the significance of differences of means and frequency of events in both groups. The level of significance was set at α = 0.05. Statistica 7.1 and Excel 2010 were used. Most smokers are in the age groups between 51-70 years. In the youngest (31-40 years), single ACF appear first (ACF <5) ACF in the number of 5-10 appear a little later (around 50 years of age) and dynamically increase, reaching a maximum at the age of 60-65.ACF in the number >10 appear at the latest age (55 years old) and their number gradually increases with age (linear growth). The probability of occurrence of ACF in all groups is greater in smokers, and the difference for the ACF group 5-10 and ACF >10 is statistically significant with a significance level of p < 0.05. Apart from ACF normal, all types of ACF are more likely in this group than in non-smokers and these differences are statistically significant with p < 0.05. CONCLUSIONS: Smoking has a significant impact on the number and type of rectal ACF. Smokers have a greater number of ACFs in the rectum than non-smokers, and the most common type is hyperplastic ACF. Smokers are more likely to develop polyps in all sections of the colon compared to non-smokers.

Effect of the Neurofeedback-EEG Training During Physical Exercise on the Range of Mental Work Performance and Individual Physiological Parameters in Swimmers.

The aim of the study was to demonstrate the effects of the Neurofeedback-EEG training during physical exercise on the improvements in mental work performance and physiological parameters. The study examined seven swimmers based on the following anthropometric measurements: body height, body mass and body composition. The Kraepelin's work curve test, EEG and EMG during physical exercise were also performed. The athletes followed 20 Neurofeedback-EEG training sessions on the swimming ergometer for 4 months. Most mean indices of partial measures of the work curve were significantly modified (p < 0.05) following the Neurofeedback-EEG training. Mean level of maximal oxygen uptake in study participants was over 55 ml/kg/min, with statistically significant differences documented between the first and the second measurements. No significant differences were found in the fatigue rate between the measurements 1 and 2. The improved mental work performance following the Neurofeedback-EEG training facilitates optimization of psychomotor activities.

Correction to: Effect of the Neurofeedback-EEG Training During Physical Exercise on the Range of Mental Work Performance and Individual Physiological Parameters in Swimmers.

The original version of the article unfortunately contained an error in the co-author details such as family name and e-mail address.

Rectal aberrant crypt foci (ACF) as a predictor of benign and malignant neoplastic lesions in the large intestine.

BACKGROUND: The importance of ACF is not fully explained, however, their number may be a good predictor of synchronous and metachronic adenoma or other polyps whose removal reduces the risk of CRC. Due to the epidemiological and genetic association of ACF with pre-cancer lesions, they may be a potential CRC biomarker. The aim of our study was to show that the number and type of rectal ACF may be a good predictive factor for the presence of polyps located proximally from the splenic flexure and that the type and number of ACF can correlate with the number and specific types of polyps in the large intestine. METHODS: The study included 131 patients who underwent colonoscopy combined with rectal mucosa staining with 0.25% methylene blue. The number of rectal ACF was determined and bioptats were sampled for histopathological examination to assess the type of ACF. Endoscopic ACF assessment criteria given by L. Roncucci were used. The obtained material was subjected to statistical analysis using probability distribution, U-test, t-student test, and chi 2 as well as the Statistica 7.1 software package. RESULTS: The study population was divided into three subgroups according to the number of ACF observed, i.e. ACF < 5, 5-10 and > 10. ACF < 5 were found in 35 patients (29.41%), 5-10 ACF in 70 (58.82%) and ACF > 10 in 14 individuals (11.76%). The study revealed the presence of normal ACF (p = 0.49), hyperplastic ACF (p = 0.34), dysplastic ACF (p = 0.11), and mixed ACF (p = 0.06). A single type of ACF was most commonly observed (n = 88, p = 0.74). In the researched group a larger number of ACF is concurrent with adenomas and hyperplastic polyps. The number of ACF clearly correlates with the dysplasia advancement in the adenoma and the number of polyps found. CONCLUSIONS: Rectal ACF are a useful marker for the presence of cancerous lesions in the proximal and distal sections of the large intestine.

Microsporidia Nosema spp. - obligate bee parasites are transmitted by air.

Microsporidia Nosema are transferred among bees via the faecal-oral route. Nosema spp. spores have been detected on flowers and transferred to hives along with the bee pollen. The aim of the present study was to determine whether Nosema microsporidia are transferred by air in an apiary, in a control area (without the presence of bee colonies), and/or in a laboratory during cage experiments with artificially infected bees. The novel way of transmission by air was investigated by the volumetric method using a Hirst-type aerobiological sampler located on the ground in the apiary, in the Botanical Garden and on the laboratory floor. Concurrently, the mean rate of Nosema infections in the foragers in the apiary was estimated with the Bürker haemocytometer method. Spore-trapping tapes were imaged by means of light microscopy, Nomarski interference contrast microscopy and scanning electron microscopy. The highest concentration of Nosema spores per 1m3 of air (4.65) was recorded in August, while the lowest concentration (2.89) was noted in July. This was confirmed by a Real-Time PCR analysis. The presence of N. apis as well as N. ceranae was detected in each of the tested tapes from the apiary. The average copy number of N. apis was estimated at 14.4 × 104 copies per 1 cm2 of the tape; whereas the number of N. ceranae was 2.24 × 104 copies per tape per 1 cm2. The results indicate that Nosema microsporidia were transferred by the wind in the apiary, but not in the Botanical Garden and laboratory by air. This was confirmed by genetic analyses. DNA from immobilised biological material was isolated and subjected to a PCR to detect the Nosema species. A fragment of the 16S rRNA gene, characteristic of Nosema apis and N. ceranae, was detected. Our research adds knowledge about the transfer of Nosema spp. microsporidia in the natural environment and indicates the season associated with the greatest risk of a bee colony infection with Nosema spp.

Breeding parameters on a mink farm infected with Aleutian mink disease virus following the use of methisoprinol.

Aleutian mink disease virus is one of the greatest threats to modern mink farming. The disease reduces fecundity and causes high mortality among kits. The aim of this study was to evaluate the effectiveness of methisoprinol in counteracting the effects of Aleutian disease, both by inhibiting replication of the virus and by mitigating the harmful effects of the disease on the fecundity and weight of infected animals. The study included 300 individuals with confirmed infection, divided according to antibody titres into three experimental groups, which received a 20% methisoprinol solution, and three control groups, which did not receive the immunostimulant. In the mink from the experimental groups, the number of copies of the genetic material of the virus in the spleens and lymph nodes was one order of magnitude lower than in the case of the control groups. Mink receiving the supplement also showed higher fecundity (on average 5.83 in the experimental groups and 4.83 in the control groups), and the weight of their offspring before slaughter was over 200 g higher. Given the lack of effective methods for immunoprophylaxis and treatment, methisoprinol supplementation can be an effective means of counteracting the effects of AMDV on persistently infected farms.

Aleutian Mink Disease Virus in the breeding environment in Poland and its place in the global epidemiology of AMDV.

Aleutian mink disease (AMD) leads to an increase in mortality of animals and causes losses in mink farming. The study investigated the presence of AMDV in tissue and environmental samples from farmed mink in Poland, and selected samples were genetically characterized. Blood, spleens and swabs from the breeding environment were collected on 27 farms in seven voivodeships in Poland (n = 250). DNA was isolated, amplified by PCR and subsequently subjected to sequencing to reveal information on the molecular epidemiology of the samples. A qPCR method was used to determine the viral load in test samples. The presence of AMDV was confirmed in tissues and the farm environment on 26 of the 27 farms. The average viral load in spleens was 108 copies. The virus was also present in the blood (average - 105 copies) and the farm environment (average - 103 copies). Isolates from the West Pomeranian Voivodeship showed high similarity within the voivodeship (over 99%). Variants from the Lublin and Podlaskie Voivodeships differed 5% from any of the AMDV isolates present in the NCBI database. Isolates from the Greater Poland, Pomeranian, Podkarpackie and Lesser Poland Voivodeships formed heterogeneous clades, showing over 97% similarity to variants previously isolated in Poland, the Netherlands and Lithuania. A high degree of genetic variation was identified among the majority of the samples, which indicates that AMDV has been introduced to Poland multiple times. However, the results within one area showed high identity between isolates, suggesting that one common ancestor was the source of these outbreaks.

A rare case of the simultaneous location of Echinococcus multilocularis in the liver and the head of the pancreas: case report analysis and review of literature.

BACKGROUND: Echinococcosis multilocularis (Hydatid Disease - HD) is a zoonotic disease caused by the larval form of Echinococcus multilocularis (EM). The main sites for this zoonosis are the Middle East, China, India, Alaska, and Siberia. It is rather rare in Europe. In Poland, the Warmian-Masurian Province is the endemic region for Echinococcus multilocularis. The clinical manifestation of the disease is dependent on the location, the size of the cyst and the development stage of the parasite. Considering the uncommon character of echinococcosis in Central Europe, especially such located in the areas outside the liver and lungs, the authors would like to present a case of coexistence in one patient of two EM foci in the liver and the head of the pancreas. CASE PRESENTATION: We present a clinical case of a 32-year-old man who was diagnosed with a cystic lesion with septa and calcification in the sixth segment of the liver and a suspicious change in the head of the pancreas. ELISA Em 2 plus test was positive, Western Blot method - the P-5 pattern showed an image that is characteristic of an EM infection. The sixth liver segment with a tumour and a tumour from the head of pancreas were excised by means of laparotomy. On the 6th day after the surgery the patient was discharged from hospital without complications and in good condition. Currently, he is under the control of a parasitic and zoonotic clinic. He takes an 800 mg daily dosage of Albendazole. CONCLUSIONS: The presented clinical case shows that if we have a patient with cystic / tumour change in the pancreas and positive immunological tests, CT and MRI of the abdominal cavity are usually sufficient in order to fully diagnose and to qualify such a person for surgery. The most effective treatment is surgical treatment supplemented with pre- and postsurgical treatment with Albendazole.