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Gastrointestinal helminths of Norway rat (Rattus norvegicus) from the Belgrade area were studied as a part of a wider ecological research of rats in Serbia (data on the distribution, population ecology, economic and epizoothiological-epidemiological importance, and density control). Rats were captured from May 2005 to July 2009 at both urban and suburban-rural sites. Of a total of 302 trapped rats 48% were males and 52% females, with 36.5% and 38.8% of juvenile-subadult individuals, per sex respectively. Intestinal helminth infection was noted in 68.5% of rats, with a higher prevalence in male hosts and in adult individuals. Higher numbers of infected juveniles-subadults were noted in suburban-rural habitats, while an opposite tendency was noted in adult rats. Seven helminth species were recovered, of which five were nematode (Heterakis spumosa, Nippostrongylus brasiliensis, Capillaria sp., Trichuris muris and Syphacia muris) and two cestode species (Hymenolepis diminuta and Rodentolepis fraterna). The most prevalent parasites were Heterakis spumosa (36.7%) and Hymenolepis diminuta (30.5%). Sex and habitat-related differences were noted in the prevalence of infection with Capillaria sp. and Trichuris muris, while there were no age-related differences in the prevalence of infection with any individual helminth species. Significantly higher prevalence of infection was noted in summer as compared to spring or winter, with a tendency to be higher in autumn as compared to spring. The only significant difference in the prevalence of infection between habitat-related was noted during spring. H. spumosa was most prevalent in summer, while H. diminuta and N. brasiliensis in autumn. The mean intensity of infection with H. spumosa, R. fraterna, S. muris and T. muris was higher in autumn than in the other seasons, while N. brasiliensis and Capillaria sp. occured in winter. No more than four helminth species were found in one host.

Warfarin (4-hydroxy coumarin) is Vitamin K (VK) antagonist that inhibits Vitamin K-dependent (VKD) processes, such as blood coagulation. It exerts influence on some non-VKD-related activities as well. In this study, the effect of subacute (30-day) oral warfarin (2 mg L-1 and 1 mg L-1) intake on hematological parameters was examined in two rat strains, Albino Oxford (AO) and Dark Agouti (DA), that differ in their sensitivity to certain chemicals. Greater susceptibility to anticoagulant effect of 2 mg L-1 of warfarin was observed in AO rats and was associated with an increase in relevant hematological parameters in this strain. Although both strains responded to 2 mg L-1 of warfarin with quantitative changes in peripheral blood leukocytes differential bone marrow and lung responses were observed. Strain-related differences in proinflammatory activity of peripheral blood granulocytes and in mononuclear cell IFN-γ production were observed. Recognition of differences in quantitative and qualitative effects of oral warfarin on processes other than hemostasis might be of relevance for those humans who are on warfarin therapy. [Projekat Ministarstva nauke Republike Srbije, br. 173039]

Background/Aim. Multiple sclerosis (MS) is an immunemediated central nervous system disease characterized by inflammation, demyelination and axonal degeneration. Cytokines are proven mediators of immunological process in MS. The aim of this study was to investigate whether there is a difference in the production of the tumor necrosis factor alpha (TNF-alpha) and interleukin-4 (IL-4) in cerebrospinal fluid (CSF) and plasma in the MS patients and the controls (other neurological non-inflammatory diseases) and to determine a possible difference in these cytokines in plasma and CSF in different clinical forms of MS. Methods. This study involved 60 consecutive MS patients - 48 patients with relapsing-remitting MS (RRMS) and 12 patients with secondary progressive MS (SPMS). The control group consisted of 20, age and sex matched, nonimmunological, neurological patients. According to the clinical presentation of MS at the time of this investigation, 34 (56.7%) patients had relapse (RRMS), 14 (23.3%) were in remission (RRMS), while the rest of the patients, 12 (20.0%), were SPMS. TNF-alpha and IL-4 concentrations were measured in the same time in CSF and plasma in the MS patients and the controls. Extended disability status score (EDSS), albumin ratio and IgG index were determined in all MS patients. Results. The MS patients had significantly higher CSF and plasma levels of TNF-alpha than the controls (p < 0.001 for both samples). IL-4 CSF levels were significantly lower in the MS patients than in the controls (p < 0.001), however plasma levels were similar. The patients in relapse (RRMS) and with progressive disease (SPMS) had higher concentrations of CSF TNF-alpha levels than the patients in remission (p < 0.001). IL-4 CSF levels in relapse (RRMS) and SPMS groups were lower than in the patients in remission. The patients in remission had an unmeasurable plasma TNF-alpha level and the patients with SPMS had significantly lower IL-4 levels in plasma than the patients in relapse and remission (p < 0.001). The only significant correlation between cytokine level with either EDSS, or albumin ratio, or IgG index, was found between CSF TNF-alpha levels and albumin ratio in the patients with relapse (R square = 0.431, p < 0.001). Conclusion. According to the obtained data MS relapse was characterized by high concentrations of TNF-alpha in CSF and plasma and low concentrations of IL-4 in CSF. Remission was characterized by high concentrations of IL-4 and low concentrations of TNF-alpha both in CSF and plasma. SPMS was characterized with lower concentrations of TNF-alpha and IL-4 compared to relapse, both in CSF and plasma.

The nematode Calodium hepaticum and the cestode Taenia taeniaeformis are zoonotic helminths primarly found in the liver of common wild rats. Most reports on these helminth species with cosmopolitan distribution are from Asia, and there is paucity of data for Europe. Wild Norway rats (Rattus norvegicus) from urban and suburban habitats of the Belgrade area were examined for the presence of Calodium hepaticum and Taenia taeniaeformis larvae liver infections. The presence of visible cysts and a histomorphology of parasite-related inflammatory liver responses were sought as signs of infection. The total prevalence of infection was 10.9% (C. hepaticum) and 29.9% (T. taeniaeformis), with no differences between the sexes. No difference in the annual prevalence of both helminth species was noted. Data obtained in this study provide new information relevant to wild Norway rats as sources of C. hepaticum and T. taeniaeformis liver infection in this geographic area, and, in a wider context, in Europe. .

Basic immune-related health-relevant changes (total and differential white blood cell counts and activity, leukocyte tissue infiltration, and related pathohistology) were assessed in wild Norway rats from urban habitats. Comparative measurements were conducted in individuals of several laboratory strains of Norway rat in order to gain insight into environmental effects on the health of wild rats. Changes in leukocyte counts and activity along with tissue infiltration were noted only in wild rats, indicating systemic as well as tissue inflammation in these animals. Coincidence of these changes with chronic inflammatory pulmonary and kidney disease was observed in the majority of affected rats.

Basic parameters of spleen immune activity (spleen weight, histomorphology of splenic compartments, and mitogen-induced splenocyte proliferative capacity in vitro) were evaluated in adult individuals of wild Norway rats from urban habitats and compared to the same data obtained in laboratory rat strains. A wider range of relative spleen mass and differential histomorphological characteristics, together with differences in the level and pattern of responsiveness of splenocytes to exogenous stimulation, were noted in spleens of wild Norway rats. Evidence of both enhanced and low-level immune-relevant spleen activity in wild rats demonstrates the complexity of changes in spleen immune activity in rats from natural populations.

The helminth fauna of the house mouse (Mus musculus Linnaeus, 1758) was studied on the basis of 429 host individuals from the suburban area of Belgrade. Eleven helminth species were recorded: three cestode species - Catenotaenia pusilla, Rodentolepis fraterna, and Cysticercus (= Strobilocercus) fasciolaris [larval stage of Taenia taeniaeformis (Batsch, 1821)]; and eight nematode species - Heligmosomoides polygyrus, Syphacia sp., Aspiculuris tetraptera, Syphacia obvelata, Heterakis spumosa, Trichuris muris, Mastophorus muris, and Gongylonema sp. Within the general helminth fauna, H. polygyrus was found to be the most prevalent species (39.2%) and caused the highest infection intensity. Prevalences of A. tetraptera, C. pusilla, and S. obvelata ranged from 12.8% to 6.1%, while the remaining species showed prevalences ranging from 4.9% (for Syphacia sp.) to 0.2% (for Gongylonema sp.). All the species found in males were also present in females, with the exceptions of M. muris and Gongylonema sp. No significant differences were found between males and females regarding prevalence (P%), mean infection intensity (MI), or mean abundance (MA).

In this study, the presence of toxigenic and pathogenic fungi was detected in individuals of natural populations of Norway rats from semiagricultural habitats. The presence of fungi was noted in 19 out of 30 (63%) individuals examined. Six fungal species were isolated and identified, of which majority belonged to Hyphomycetes (Deuteromycotina) and Mucor racemosus from Zygomycotina. All of detected species are of public health importance and some of them might influence animals health.

In this study, histopathology of local pulmonary response following intranasal inoculation of different doses of Aspergillus fumigatus conidia in laboratory rats was evaluated. Development of response was evaluated on days seven and twenty one, following conidia inoculation by cell infiltration and by presence of A. fumigatus conidia in homogenates of lung tissue. Total and differential peripheral blood leukocyte counts and state of leukocyte adhesion/aggregation were monitored to estimate the presence of systemic response in infected individuals. Mycological examination revealed the presence of conidia in lung tissue homogenates of infected animals, with high number of non-germinating spores on day twenty one, following the inoculation of lower conidia dose. Histopathological examination revealed the presence of lymphocytes perivascularly and in vascular lumen in the lungs. Presented data demonstrate pulmonary immune response following A. fumigatus conidia administration.

In this study systemic response to intravenous administration of Aspergillus fumigatus conidia was investigated. The intensity of response was evaluated by a survival rate and by histopathological tissue analysis. Administration of all doses (106 - 5x107) of Aspergillus fumigatus conidia caused mortality, but the highest mortality and the shorter time of survival were noted at higher doses applied. At the highest applied dose, the presence of spores and hyphae was noted in lungs and kidneys. Histological analysis revealed the presence of intense inflammatory reaction in lungs, kidneys and spleen. Functional and histological changes observed provide means to study both mechanisms and drug interventions in systemic Aspergillus infection.

Background. Cytokines are involved in almost every aspect of reproduction and recent studies suggested a relationship between cytokines and male/female infertility. In the present study, the levels of interleukin-8 (IL-8) and interleukin-1 receptor antagonist (IL-1Ra) were determined in the cervical mucus of fertile and infertile women. Methods. Groups of patients were formed according to the results of the standard procedure for infertility investigation, including postcoital test and the presence of antispermatozoid antibodies in the sera of both partners and in seminal plasma, by mixed antiglobulin reaction (MAR) test and Kibrick agglutination test. IL-8 and IL-1Ra levels were determined in solubilized (ultrasonographic sonicaton) cervical mucus sampled in the midcycle by commercial ELISA kits and expressed as pg/mg proteins. Results. The groups were designated as fertile (n=20) and infertile (n=48). The latter was divided into two subgroups, one consisting of infertile women with positive postcoital test and without antispermatozoid antibodies (n=30), and the other designated as infertile women with negative postcoital test (n=18). This subgroup was composed of women with negative postcoital test and without antibodies (n=10) and the women with negative postcoital test with antibodies (n=8). Similar levels of IL-8 and IL-1 Ra were noted in the cervical mucus of in fertile women and women with positive postcoital test and without antispermatozoid antibodies. A tendency of decrease (p=0,052) and significant decrease in IL-8 levels (p

Background. Contact hypersensitivity (CHS) is a local inflammatory response of the skin following challenge of hapten-sensitized animals. It is the consequence of cell infiltration of derm and the release of inflammation mediators, among which Tumor necrosis factor-alpha (TNF-α) is one of the most important factors. The intensity of the inflammation could be quantified by ear swelling which is the classical manifestation of the reaction. This study was testing the working hypothesis that levels of TNF-α in skin organ culture medium should correlate with the intensity of CHS reaction measured in vivo by ear swelling assay, and with the density of dermal infiltrate in ear skin samples. In order to test the working hypothesis, the intensity of inflammatory reaction following challenge was evaluated by classical measurements of ear swelling, by the determination of TNF-α levels in culture fluids of ear skin following epicutaneous application of dinitrochlorobenzene (DNCB) into the ears of sensitized animals. Methods. Animal model of CHS reaction to DNCB in Albino Oxford rats was used as described. Ear swelling was quantified in percentage terms as the difference in thickness between the challenged and nontreated ears of the same animal. Dermal infiltrate density in histopathologically analyzed samples of ear skin was evaluated by computer-assisted image analysis. Ear skin samples were cultured in standard medium for 24 h, and TNF-α concentration in the conditioned medium was subsequently determined with ELISA test. Results. Dose-dependent increase in the density of the dermal infiltrate and in TNF-α in CM were noted following the application of 0.65%, 1.3% and 2.6% of DNCB to the ears of previously sensitized rats. The correlation between ear swelling and the levels of TNF-α (r=0.933, p