Post-harvest Salmonella spp. prevalence in turkey carcasses in processing plant in the northeast part of Poland.

Turkeys carcasses at selected point after slaughter on dressing line in poultry were sampled and analyzed for Salmonella. These slaughter turkeys came from the northeast part of Poland. The examinations were carried out in each month of 2009. Three hundred turkeys were selected at random from a commercial slaughter line, immediately after completing the cooling process. The percentage of these 300 turkeys from which Salmonella spp. were isolated was relatively high (8.3%; Salmonella positive results were observed in 25 cases). The lowest Salmonella spp. rate (1.3 %) for slaughter birds was found in the fourth quarter, and the highest contamination rate at 18.6% was found, in the third quarter. The serological types of Salmonella spp. isolated from the whole turkey carcasses were S. Saintpaul, S. Senftenberg, S. Anatum, S. Heidelberg, S. Hadar, S. Typhimurium and S. Infantis.

Seronegative hepatitis C virus infection in patients with lymphoproliferative disorders.

It has been reported that hepatitis C virus (HCV) RNA may be present in serum and/or lymphoid cells in the absence of specific circulating antibodies. The current study analysed seronegative HCV infection in patients with lymphoproliferative disorders. We studied 77 anti-HCV-negative patients (45 male and 32 female, mean age 54.8 ± 14.2 years) with various lymphoproliferative disorders. HCV-RNA was detected by RT-PCR in plasma, peripheral blood mononuclear cells (PBMC) and bone marrow. Furthermore, the presence of viral nonstructural protein 3 (NS3) was determined in PBMC and bone marrow by immunostaining. HCV-RNA was detectable in at least one compartment in 27 (35.1%) patients. Viral RNA was found in bone marrow in 22 patients (28.6%), in PBMC in 13 (16.9%) and in plasma in 10 (13%) patients. In nine patients, evidence of infection was confined to the bone marrow compartment. Viral load in HCV-RNA-positive plasma ranged from 15 to 1.17 × 10(3) IU/mL. NS3 was detected in all but two HCV-RNA-positive bone marrow samples and in all but one HCV-RNA-positive PBMC samples. All 27 HCV-RNA-positive patients remained anti-HCV-negative when tested again after 6-12 months, but only four remained HCV-RNA positive. In conclusion, among patients with lymphoproliferative disorders, HCV can be present in plasma, PBMC and bone marrow despite the lack of circulating specific antibodies. Further studies are required to analyse the phenomenon of seronegative infection and to determine whether such patients are infectious.

Hepatitis C virus 5' untranslated region variability correlates with treatment outcome.

Hepatitis C virus (HCV) variability affects viral-host interactions. We analysed HCV 5'untranslated region (5'UTR) in sera and peripheral blood mononuclear cells (PBMC) from chronic hepatitis C patients undergoing antiviral treatment. We studied 139 patients treated with pegylated interferon and ribavirin. The primary endpoint was a sustained virological response (SVR) defined as negative HCV RNA level 24 weeks after the end of therapy. 5'UTR was analysed by single-strand conformational polymorphism (SSCP) and sequencing. The pretreatment SSCP pattern in serum and PBMC differed in 26 (18.7%) patients. During therapy, the SSCP pattern remained stable in 65 (60.8%) patients, number of bands declined in 16 (15.0%), and in 18 (16.8%) patients, changes were qualified as 'shift' indicating change in band positions. In univariate analysis, there was a significant (P ≤ 0.05) positive association between SVR and pretreatment serum and PBMC dissimilarities, initial viral load <10(6) IU/mL, IL-28B CC genotype of the rs12979860 single nucleotide polymorphism and change in the SSCP band pattern (either 'shift' or decline) In multivariable analysis, only low initial viral load, IL-28B genotype, and changes in the SSCP band pattern were independent factors associated with SVR. In conclusion, stability of 5'UTR correlated with infection persistence, while changes correlated with SVR.

Variability of hepatitis C virus hypervariable region 1 (HVR-1) during the early phase of pegylated interferon and ribavirin therapy.

PURPOSE: Genetic variability of hepatitis C virus (HCV) is considered to be an important factor defining viral pathogenesis, persistence and resistance to treatment. The aim of the present study was to characterize HCV genetic heterogeneity within a hypervariable region 1 (HVR-1) before and during the early period of pegylated interferon alfa (PEG-IFN-α) and ribavirin treatment in correlation with treatment outcome. MATERIAL AND METHODS: The study involved 24 patients treated with PEG-IFN-α and ribavirin whose sera were collected before (baseline) and at 7, 14, 21 28 and 56 day of treatment. HCV HVR-1 region was amplified by nested RT- PCR and subjected to SSCP (single strand conformational polymorphism) analysis. SSCP changes of HCV HVR-1 over time in each patient were compared to treatment outcome results. RESULTS: In 2/11 (18%) SVR+ and 8/13 (62%) SVR- treated patients, HVR-1 genetic changes manifested by new SSCP bands (new genetic variants) and were significantly more frequent in nonresponders (P <0.05). CONCLUSIONS: Our results indicate that HCV HVR-1 variability during the early phase of PEG-IFN-α and ribavirin therapy may be predictive of treatment outcome.

Neoplastic lesions in slaughter animals in Warminsko-Mazurskie voivodship (Poland) area during the years 2001-2007.

During the period of 7 years, 2001-2007, 110,273 heads of cattle and 3,055,654 heads of pigs were examined by veterinarians of the Polish Veterinary Inspection in Warminsko-Mazurskie voivodship. As a result of pre- and post-slaughter examination, 311 heads of cattle and 23 heads of pigs representing 0.279% and 0.00075% of the population respectively were considered unsuitable due to neoplastic lesions. The most frequently found lesions in cattle were fibroneuromas, neuromas and liposarcomas while in case of pigs those were various types of sarcomas, fibromas, cancers, nephromas and melanomas.

Selection of different 5' untranslated region hepatitis C virus variants during post-transfusion and post-transplantation infection.

BACKGROUND: Hepatitis C virus (HCV) translation is initiated in a cap-independent manner by an internal ribosome entry site (IRES) located within the 5' untranslated region (5'UTR). Sequence changes in this region could affect translation efficiency and presumably viral replication. AIM: To determine translation efficiency of 5'UTR variants developing during post-transfusion hepatitis C in two immunocompetent subjects and in two immunosuppressed liver recipients with recurrent HCV. METHODS: Sequential samples were screened for 5'UTR changes by single-strand conformation polymorphism followed by cloning and sequencing whenever band pattern suggested sequence changes. 5'UTR variants were tested for IRES activity using a bicistronic dual luciferase expression plasmid transfected into HepG2 and Huh7 cell-lines. RESULTS: In the transfused patients, translation efficiency of 5'UTR variants from early post-transfusion samples was 5.1- to 13.7-fold higher than that of predominant variants found in late follow-up samples. Post-transplant variants in the other two patients had 2.6- to 5.9-fold higher translation efficiency than those present only in pretransplant samples. CONCLUSION: In the immunocompetent host there may be selection of low translation efficiency HCV variants over the course of infection. However, in immunosuppressed subjects the opposite seems to be true as low translation efficiency variants are superseded by high translation efficiency variants.

The effect of polyphosphates on streptococci isolated from mastitis cases.

The effect of 0.01, 0.02, 0.03, 0.05, 0.6, 0.8 and 0.9% of polyphosphate mixture on 5 strains of Streptococcus agalactiae, 2 strains of Streptococcus dysgalactiae and 3 strains of Steptococcus uberis in blood agar in vitro and in milk was examined. It was found that the growth of all strains of the above mentioned bacteria was completely inhibited by 0.03% phosphate mixture in blood agar. In milk, however, the amount of polyphosphate mixture necessary for total inhibition was, from 0.6 to 0.9%, depending on the strain.

The legal requirements concerning transport of slaughter animals in Poland.

Conditions of trading and handling of animals during transportation are regulated by numerous norms which cover the humanitarian, sanitary, veterinary as well as technical and construction requirements to transportation means. Veterinary services play a particular role in trade of slaughter animals due to their controlling authority. They execute sanitary supervision over animal trading according to the regulations. Polish legislation related to the transport of animals is adjusted to the European Union standards.

Natural history of acute symptomatic hepatitis type C.

BACKGROUND: Spontaneous clearance of hepatitis C virus (HCV) after acute hepatitis C, and the course of chronic HCV infection in patients who did not clear the virus, were studied. PATIENTS AND METHODS: Patients with acute C or non-A, non-B hepatitis who were hospitalized between 1988 and 1998 were called for evaluation in 2001. They were tested for anti-HCV, serum HCV-RNA, HCV-RNA in peripheral blood mononuclear cells (PBMC) and liver enzymes. A liver biopsy was performed on chronically infected patients. The course of acute hepatitis C was compared between HCV-RNA-positive and negative subjects to look for factors that might influence spontaneous viral clearance. Factors influencing more progressive liver disease were analyzed in chronic hepatitis C. RESULTS: Out of 159 acute hepatitis C patients, 77 (48.4%) participated in the study, and the median observation time was 8 years. Spontaneous clearance of serum HCV was found in 23 subjects (29.9%), but in two cases HCV-RNA was detected in peripherical blood mononuclear cells (PBMC). Only three patients negative for HCV-RNA in serum and PBMC lost anti-HCV. Severity of acute HCV infection and previous alcohol abuse seemed to influence resolution. In non-alcoholic patients, older age at time of primary infection was a significant predictor of virus clearance. In chronic hepatitis C, more than 75% of patients had minimal or mild activity in biopsy, but 40% had advanced fibrosis. Older age at infection, male gender, alcohol abuse, and higher iron content were connected with advanced fibrosis. CONCLUSION: Studies on HCV infection resolution should include at least PBMC testing for HCV-RNA. A healthy carrier state of HCV can be discussed. A longer observation time increased the likelihood of seroreversion. Fibrosis in chronic hepatitis C probably is not a direct result of inflammatory activity.

Elimination of Salmonella spp. by lactic acid.

The aim of the present study was to determine the elimination of Salmonella by different lactic acid concentrations in microbiological media and on turkey carcass elements. The average bacteria counts in the control samples without lactic acid were: 1.8 x 10(8), 1.1 x 10(8) and 2.3 x 10(8), for S. Enteritidis, S. Anatum and S. Typhimurium, respectively. The concentration of lactic acid of 0.1% in the agar media completely inhibited the growth of all Salmonella strains. At 0.05% lactic acid concentration, the bacteria count was 2 log cycles lower and at a 0.03% solution it was 1 log cycle lower than that in the respective control samples. However, the examined bacteria developed in the presence of 0.02% and 0.01% lactic acid concentrations and their counts fell into the same log brackets. An analysis of the experimental results obtained from turkey carcass elements immersed in the lactic acid solution showed that the Salmonella identification rate was determined by the bacteria inoculum spread over the turkey carcass surface. The contamination of 10(1) CFU of Salmonella spread onto the turkey carcass was completely eliminated by immersing the carcasses in 1% or 2% lactic acid solutions. The contamination of turkey carcass elements with 10(2) CFU of S. Enteritidis and their immersion in 2% lactic acid solution for 15 min resulted in the reduction of the number of samples with Salmonella compared to the number of control samples with Salmonella. At contaminations of 10(3) CFU on the carcass surfaces, the immersions in 1% and 2% lactic acid solutions did not reduce Salmonella counts.

Changes in hepatitis C virus population in serum and peripheral blood mononuclear cells in chronically infected patients receiving liver graft from infected donors.

BACKGROUND: We have previously studied hepatitis C (HCV)-infected recipients of livers from HCV-infected donors and found that either the donor's strain or the recipient's strain predominate in serum. The current study was undertaken to determine whether these changes are complete and whether they are reflected in the population of virus associated with peripheral blood mononuclear cells (PBMCs). METHODS: We analyzed HCV ribonucleic acid from sequential serum and PBMC samples from 11 and 8 patients, respectively. The relatively stable NS5 region was chosen for analysis because it allowed for dependable identification of donor and recipient strains. Viral sequences were analyzed by direct sequencing and by sensitive strain-specific polymerase chain reaction assays. These assays were capable of detecting the minor sequence present at a concentration 1:104-10-7 below that of the major sequence. RESULTS: Five patients retained their original infecting strain; the donor strain was detected only transiently. In the remaining six patients, recipient strain was detected for the first few weeks, after which only the donor strain was consistently present. However, in one patient the second nondominant strain was detected from the background of the major strain on a single occasion 8 months after transplantation. All changes in serum were closely paralleled by those occurring in PBMCs. CONCLUSIONS: Viral population changes in the setting of liver transplantation from HCV-infected donors to HCV-infected recipients occur simultaneously in PBMCs and serum. The takeover of one strain by another in PBMC- and serum-derived viral populations seemed to be complete and long lasting.

Occurrence of Salmonella spp. in consumption eggs in Poland.

The purpose of this study was to determine the prevalence of Salmonellae on egg shells in markets in Olsztyn, Poland. An investigation carried out by the Sanitary and Epidemiological Station into cases of food poisoning caused in Poland by Salmonella spp. in recent years showed that the largest number of outbreaks is connected with consuming foods containing hens eggs which had not undergone heat treatment, such as mayonnaise, creams, ice-cream and other products. The world egg production amounts to 400 billion, in Poland it reaches the level of around 8 billion per year. A total of 1200 eggs were purchased in 40 local markets in Olsztyn were examined for the presence of Salmonella between June 1997 and December 1998. Salmonella was not found on the shell or inside the eggs. From this study it would appear that the incidence of Salmonellae on eggs from Olsztyn shops is very low.

Exposure of hepatitis C virus (HCV) RNA-positive recipients to HCV RNA-positive blood donors results in rapid predominance of a single donor strain and exclusion and/or suppression of the recipient strain.

We have analyzed three cases of hepatitis C virus (HCV)-infected recipients who received blood from HCV-infected donors. Two recipients were exposed to two different HCV RNA-positive donors, and one was exposed to a single donor. All parental genomes from the actual infecting units of blood and the recipients were defined, and their presence in the follow-up serum samples was determined using sensitive strain-specific assays. The strain from one of the donors was found to predominate in all recipients' serum samples collected throughout the follow-up period of 10 to 30 months. In two recipients exposed to two infected donors, the strain from the second donor was occasionally found at very low level. However, the original recipients' strains were not detected. Our observations show that HCV-infected individuals can be superinfected with different strains, and this event may lead to eradication or suppression of the original infecting strain. Furthermore, our findings demonstrate that simultaneous exposure to multiple HCV strains may result in concomitant infection by more than one strain, although a single strain could rapidly establish its dominance. The results of the present study suggest the existence of competition among infecting HCV strains which determines the ultimate outcome of multiple HCV exposure.

Exposure of hepatitis C virus-negative recipients to > or =2 infected blood donors.

This study analyzed 4 cases of hepatitis C virus (HCV)-naive transfusion recipients who developed hepatitis after receiving blood from >1 HCV-infected donor. One recipient was exposed to 4 donors, 2 were exposed to 3 donors, and 1 was exposed to 2 donors. For 3 recipients, the strain from 1 of the donors predominated in all follow-up samples collected for 8-40 months. For 2 recipients, the strain from the second donor was occasionally detectable with sensitive strain-specific assays. For the fourth recipient, the initially dominant strain was later supplanted by a strain from the other donor. Simultaneous exposure to multiple HCV strains may result in concomitant infection by >1 strain, although a single strain rapidly establishes its dominance. These observations are compatible with the presence of competition among infecting HCV strains that results in the dominance of 1 strain and competitive exclusion or suppression of other strains.

Hypertonicity of dialysis fluid suppresses intraperitoneal inflammation.

We studied acute and chronic intraperitoneal inflammation during dialysis performed in rats injected with phosphate-buffered saline alone (PBS) or PBS supplemented with glucose (Glu) or with mannitol (Man). In acute experiments, the result of a first dialysis with PBS in every rat (dialysis I) was compared with a second dialysis performed 24 hours later (dialysis II) using a different dialysis fluid: either PBS with 3.86 g/dL Man or PBS with 3.86 g/dL Glu. In rats exposed to hypertonic dialysis solutions (both Glu and Man), inflammatory reaction was decreased (cell count: p < 0.05; nitric oxide secretion: p < 0.05; protein in dialysate: p < 0.05). In control animals treated only with PBS (dialysis I and dialysis II), the inflammatory reactions during dialysis I and dialysis II were comparable. In chronic experiments, rats were dialyzed with the tested fluids for four weeks. Weekly, dialysate samples were taken and analyzed. At the end of the study, cell counts and protein losses were higher in the PBS-treated rats than in the other groups (cell count: p < 0.05, Glu vs PBS, and p < 0.05, Man vs PBS; protein in dialysate: p < 0.001, Glu vs PBS, and p < 0.01, Man vs PBS). We conclude that hypertonicity of the dialysis fluid inhibits the inflammatory reaction in the peritoneal cavity during peritoneal dialysis.

Detection of active hepatitis C virus and hepatitis G virus/GB virus C replication in bone marrow in human subjects.

We have analyzed the presence of hepatitis C virus (HCV) and hepatitis G virus (HGV) sequences in bone marrow and serum samples from 48 patients of a hematologic outpatient clinic. HCV RNA was detected in 18 (38%) and 15 (31%) and HGV RNA was detected in 6 (13%) and 9 (19%) of serum and bone marrow samples, respectively. In 3 patients, HGV RNA was detectable in bone marrow but not in the serum; 2 of these patients were negative for the presence of specific antibodies. Using a highly strand-specific Tth-based reverse transcriptase-polymerase chain reaction (RT-PCR), the presence of HCV RNA and HGV RNA negative strand was demonstrated in 4 and 5 bone marrow samples, respectively. Our study shows that HCV and HGV can replicate in bone marrow; in the case of HGV, analysis of serum may underestimate the true prevalence of infection. (Blood. 2000;95:3986-3989)

Hepatitis C virus in lymphoid cells of patients coinfected with human immunodeficiency virus type 1: evidence of active replication in monocytes/macrophages and lymphocytes.

It has been reported that hepatitis C virus (HCV) may be lymphotropic in the setting of human immunodeficiency virus type 1 (HIV-1) coinfection. The present study was undertaken to determine the phenotype of lymphoid cells harboring replicating HCV in HIV-1-positive subjects. By means of highly strand-specific thermostable enzyme Tth-based reverse-transcriptase polymerase chain reaction, the presence of viral RNA-negative strand was sought in different subpopulations of peripheral blood mononuclear cells from 10 HIV-positive patients. HCV RNA-negative strand was most commonly present in monocytes/macrophages (4 cases), followed by CD8+ and CD4+ lymphocytes (2 cases) and CD19+ cells (1 case). In 2 cases that were further analyzed, viral-negative strand remained detectable in monocytes/macrophages cultured for 3 weeks. Moreover, monocyte/macrophage- and serum-derived viral sequences differed in the 5' untranslated region. These findings imply that, in HIV-infected subjects, HCV may replicate in the same cells as HIV-1, which raises the possibility of direct interactions between these pathogens.

Uneven distribution of hepatitis C virus quasispecies in tissues from subjects with end-stage liver disease: confounding effect of viral adsorption and mounting evidence for the presence of low-level extrahepatic replication.

We have found differences among the populations of hepatitis C virus sequences in serum, peripheral blood mononuclear cells (PBMCs), and various tissues in patients with chronic hepatitis C. These results are compatible with the existence of independent viral compartments in the infected host. Our results also suggest that PBMCs, and probably various tissues, can selectively adsorb viral subpopulations differing in the E2 region.