An IgM murine monoclonal antibody (MAb) was obtained against the excretory-secretory antigen (ES-Ag) of in vitro reared protoscoleces of Echinococcus granulosus (Batsch, 1786). Western blotting revealed that the MAb recognised a 20.6 kDa protein of this ES-Ag. The MAb was used in sandwich enzyme-linked immunosorbent assay (s-ELISA) for selective sensitisation of the solid phase with the protoscolex-specific protein from its ES-Ag and somatic antigen (S-Ag) to examine serum samples of 108 cows from a cystic echinococcosis (CE) endemic area for specific antibodies and to compare the results with those from necropsies and an indirect ELISA (i-ELISA). The sensitivity of s-ELISA/ES-Ag, s-ELISA/S-Ag and i-ELISA/S-Ag was 48%, 52% and 62%, respectively. The low sensitivity of the ELISA was probably caused by the fact that 13 cows (62%) were infected with sterile cysts (acephalocysts and/or calcified foci) only. A relatively high specificity (80%) of s-ELISA/ES-Ag was observed in cows with fertile cysts. It also detected antibodies in the serum of two cows that had recovered from the disease according to the necropsy. The i-ELISA/S-Ag gave false results in testing sera from a healthy animal and from a cow with tubercular foci. Further analysis will be necessary to define more precisely the value of this study, because the duration of antibody elimination from the bloodstream of recovered cattle remains unknown. The solution of this problem will increase the specificity of the proposed test in monitoring herbivorous animals for CE., Aitbay K. Bulashev, Zhanbolat A. Suranshiev, Orken S. Akibekov, Zhanara Zh. Akanova, Gulmira A. Abulgazimova., and Obsahuje bibliografii
Hunting dogs have probably a higher level of exposure to Neospora caninum Dubey, Carpenter, Speer, Topper et Uggla, 1988 and Toxoplasma gondii Nicolle et Manceaux, 1908 than other canine populations for their different lifestyle. The aim of our survey was to determine the seroprevalence of N. caninum and T. gondii in hunting dogs from southern Italy and assess risk factors related to these protozoan infections. Blood samples were collected from 398 hunting dogs (19 different breeds, aged from 5 month to 14 years). The sera were screened by indirect fluorescence antibody test; a titre ≥ 50 was considered positive. Antibodies to N. caninum and T. gondii were detected in 59 (15%) dogs with titres from 50 to 3 200 and in 94 (24%) dogs with titres from 50 to 1 600, respectively, with co-infection in 25 (6%) dogs. Statistical difference (p ≤ 0.05) was found only for infection with T. gondii between two age groups: ≥ 2-4 years (16%) and ≥ 4-7 years (33%); other observed characteristics were without statistical significance. Our results suggest that the hunting dogs could play an important role in the transmission cycle of N. caninum between wild animals and livestock. This is the first detection of antibodies to T. gondii in hunting dogs in Italy., Tereza Machačová, Eva Bártová, Kamil Sedlák, Radka Slezáková, Marie Budíková, Diego Piantedosi, Vincenzo Veneziano., and Obsahuje bibliografii
The consumption of horse meat has been epidemiologically linked to clinical toxoplasmosis in humans and neosporosis that may cause clinical illness in horses. Here we determined seroprevalence of antibodies against Toxoplasma gondii Nicolle et Manceaux, 1908 and species of Neospora Dubey, Carpenter, Speer, Topper et Uggla, 1988 in horses from Italy. Blood samples were collected from 643 apparently healthy horses from 60 farms of 51 municipalities in southern Italy. The presence of antibodies against T. gondii and Neospora spp. were detected by indirect fluorescence antibody test (IFAT); a titre ≥ 50 was considered positive. The same sera were also tested for antibodies against Neospora spp. by a competitive-inhibition enzyme-linked immunosorbent assay (cELISA); samples with ≥ 30% inhibition were considered positive. Antibodies against T. gondii and Neospora spp. were detected in 19 (3.0%) and 15 (2.3%) horses by IFAT, respectively, without statistical difference between gender, age and breeds (p-value ≥ 0.05). Antibodies against species of Neospora were detected in 70 (10.9%) horses by cELISA with statistical difference in gender (6.0-18.5%, p-value ≥ 0.05) and breeds (0-19.4%, p-value ≥ 0.05). Although T. gondii infection rates were low, the risk of human infection should not be dismissed, particularly in Italy where consumption of raw or undercooked horse meat has a long tradition., Eva Bártová, Tereza Machačová, Kamil Sedlák, Marie Budíková, Ugo Mariani, Vincenzo Veneziano., and Obsahuje bibliografii
Toxoplasma gondii (Nicolle et Manceaux, 1908) is an obligate intracellular, parasitic protozoan within the phylum Apicomplexa that causes toxoplasmosis in mammalian hosts (including humans) and birds. Since meat of wild boar, Sus scrofa (Linnaeus), has been demonstrated to be a potential source of human infection, a careful evaluation of the prevalence of infection with T. gondii in hunted animals is needed to protect public health. In the Var area in southeastern France, we performed a spatio-temporal survey in order to investigate the prevalence of IgG antibodies in wild boars shot by hunters in the Canjuers military camp during two subsequent hunting seasons. Of 841 wild boars screened, antibodies (IgG) to T. gondii (modified agglutination test, cut-off 1 : 6) were found in 141 (16.8%) muscle extract samples. A significant association (p < 0.001) was found between positivity and age, but not gender, and hunting districts. The results obtained indicate that consumption of raw or undercooked meat from wild boars carries an important risk of infection with T. gondii. Wild boars may be considered as a bioindicator of parasite circulation in this ecosystem., Cédric Roqueplo, Radu Blaga, Jean-Lou Marié, Isabelle Vallée, Bernard Davoust., and Obsahuje bibliografii
Toxoplasma gondii (Nicolle et Manceaux, 1908) is an obligatory intracellular protozoan parasite prevalent in animals and humans worldwide having medical and veterinary importance on account of causing abortion or congenital disease in intermediate hosts, including man. Since T. gondii has already been identified in the milk of goats, Capra aegagrus hircus (Linnaeus), the possibility of acquiring infection by ingesting unpasteurised goat milk should be taken into consideration. Thus, the aim of the present study was to determine the presence of T. gondii DNA in goat milk. First, 73 goats (females) from 36 farms located in Poland were examined serologically by direct agglutination test (DAT) to estimate the T. gondii serological status. Milk samples from 60 selected lactating females were examined for the presence of T. gondii DNA by Real time PCR and nested PCR (B1 gene). To estimate the clonal type of detected T. gondii, multiplex PCR was performed using 6 markers. In DAT, positive results were found in 70% of 73 goats. Among examined 60 milk samples, 65% were positive in Real time PCR and 43% in nested PCR. It is noteworthy that 11 samples positive in PCR were collected from seronegative goats. The multilocus PCR analysis mostly revealed the occurrence of genotype III, which is relatively rare in Europe. The recorded high prevalence of anti-Toxoplasma antibodies in tested goats (70%), associated with a high prevalence of T. gondii DNA in goat milk samples (65%), indicates a potential risk of the parasite transmission through goat milk ingestion., Jacek Sroka, Paweł Kusyk, Ewa Bilska-Zając, Jacek Karamon, Jacek Dutkiewicz, Angelina Wójcik-Fatla, Violetta Zając, Krzysztof Stojecki, Mirosław Różycki, Tomasz Cencek., and Obsahuje bibliografii
A nonpigmenting strain of Serratia marcescens Bizio isolated from dead and apparently diseased wild apple maggot flies, Rhagoletis pomonella (Walsh), was shown to be pathogenic to healthy apple maggot flies upon ingestion. The microorganism was detected in live adult alimentary canal organs four days post ingestion but produced death in some flies within 24 h when flies fed on a cell concentration of 4.7 × 104 cfu/ml and within 8 h when flies fed on filter-sterilized culture medium that previously contained a 21 h culture of S. marcescens. Increasing the cell concentration 10,000 fold did not lead to an increased rate of kill. Young flies (7-10 days old) were more susceptible to infection leading to death than were older flies (21-28 days old). The potential use of S. marcescens cells as control agents against apple maggot flies is negated by their pathogenicity to vertebrates; however, the potential use of toxic compounds produced by this strain of S. marcescens is discussed.
Five intratesticular sex steroids (testosterone, dihydrotestosterone, androstenedione, estradiol and epitestosterone) along with six serum hormones (LH, FSH, prolactin, SHBG, testosterone and estradiol) were determined in 84 non-obstructive azoospermic men, in order to evaluate to what extent serum and testicular tissue as well as individual hormones in the same material mutually correlate. With exception of androstenedione, tight correlations were found among tissue content of sex steroids, while only weak correlation was recorded between serum and testicular concentrations of major sex steroids testosterone and estradiol. It points to importance of measurement of intratesticular steroids in combination with examination of sperm parameters for assessment of testicular function and spermatogenesis., J. Heráček, V. Sobotka, L. Kolátorová, J. Kočárek, R. Hampl., and Obsahuje bibliografii
Hepcidin is a key regulator of iron metabolism and plays an important role in many pathologies. It is increased by iron administration and by inflammation, while erythropoiesis downregulates its expression. It decreases iron availability and thus contributes to anemia of chronic diseases. The aim of the study was to measure hepcidin as a marker and pathogenetic factor in ANCA-associated vasculitis (AAV). Hepcidin plasma concentration was measured by the immunological method in 59 patients with AAV and compared to patients with non-vasculitic etiology of chronic kidney disease, patients on hemodialysis (HD), with systemic lupus erythematodes (SLE) and to healthy controls and blood donors, and was correlated with the parameters of iron metabolism, inflammation, activity of the process and kidney function. Hepcidin concentration was increased in patients with AAV, SLE and HD and correlated positively with C-reactive protein, serum ferritin and creatinine, and negatively with hemoglobin and serum transferrin. In active form of AAV it correlated with the clinical scoring system (BVAS). Hepcidin can thus be considered as a pathogenetic factor of anemia in AAV and can be used for evaluation of inflammation in AAV and as an additional marker in active forms of the disease.
High-mobility group box 1 (HMGB1) is newly discovered protein, which play a crucial role in the pathogenesis of systemic inflammation. Recent studies showed that HMGB1 is one of the important pathophysiological mechanisms in the occurrence and development of atherosclerosis. The purpose of the present study was to investigate the relationship between serum HMGB1 levels and endothelial function in patients with polycystic ovary syndrome (PCOS). Eighty newly diagnosed patients with PCOS and eighty normal women of similar age were selected. Metformin treatment (1,500 mg/day) was initiated in all patients for a period of consecutive 3 months. Serum HMGB1 levels were measured by ELISA. High resolution ultrasound was used to measure the brachial artery diameter at rest, after reactive hyperemia (flow-mediated arterial dilation, FMD) and after sublingual glyceryltrinitrate. Serum HMGB1 levels in PCOS were 24.87±14.93 ng/ml, which were significantly higher than that in controls (8.82±3.55 ng/ml, p<0.01). After 3 months treatment, serum HMGB1 levels decreased significantly (p<0.05). By dividing the distribution of HMGB1 levels into quartiles, serum HMGB1 levels were increased gradually with the increase of testosterone levels (p<0.05), whereas the FMD levels decreased (p<0.05). Multiple stepwise linear regression analysis showed that FMD (estimated coefficient β=-0.69, p=0.005), testosterone (β=0.31, p=0.045), TBARS (β=0.69, p=0.012) and hs-CRP levels (β=0.68, p=0.001) were significantly associated with HMGB1. The absolute changes in HMGB1 showed a positive correlation with the changes in testosterone (p<0.05) and negative correlation with the changes in FMD (p<0.05) in patients with PCOS during the course of metformin therapy. Serum HMGB1 levels are correlated with endothelial dysfunction in patients with PCOS. Our study suggests that HMGB1 may contribute to the early stage of atherosclerosis in patients with PCOS.