Differential Activity of Human Leukocyte Extract on Systemic Immune Response and Cyst Growth in Mice with Echinococcus Multilocularis Infection After Oral, Subcutaneous and Intraperitoneal Routes of Administration.

Alveolar echinococcosis (AE) caused by the larval stage of Echinococcus multilocularis is serious parasitic diseases associated with the host´s immunosuppression. The effects of human non-immune dialyzable leukocyte extract (DLE) on immune cells in blood and spleen and parasitic cysts weight in Balb/c mice after oral (PO), subcutaneous (SC) and intraperitoneal administration (IP) were compared. The reduction in cysts weight (p < 0.01) was recorded after PO route, whereas moderate reduction was found after SC and IP routes. The elevation of lymphoid populations in blood and spleen was found after PO administration (p < 0.01) in parallel with reduced myeloid population. Infection-elicited decline in B220+B cells was partially abolished by PO route, but DLE routes did not influence the CD3+ T cells. The proportions of CD3+CD4+Th lymphocytes were moderately upregulated, whereas CD3+CD8+Tc populations were reduced after all DLE routes (p < 0.01). PO administration increased CD11b+MHCIIhigh blood monocytes, CD11b-SigleF+ cell, but not CD11b+Si-glecF+ eosinophils in the blood, stimulated after SC and IP routes. DLE induced downregulation of NO production by LPS-stimulated adherent splenocytes ex vivo. Con A-triggered T lymphocyte proliferation was associated with the elevated IFN-γ production and transcription factor Tbet mRNA expression. The alleviation of Th2 (IL-4) and Treg (TGF-ß) cytokine production by lymphocytes ex vivo paralleled with downregulation of gene transcription for cytokines, GATA and FoxP3. Reduction of myeloid cells with suppressive activity was found. The SC and IP routes affected partially the cysts weights, diminished significantly gene transcription, NO levels and Th2 and Treg cytokines production. Results showed that PO route of DLE administration was the most effective in ameliorating immunosuppression via stimulation of Th1 type, reducing Th2 and Treg type of immunity and CD3+CD8+Tc lymphocytes in the blood and spleens during E. multilocularis infection in mice.

Probiotic Bacteria can Modulate Murine Macrophage's Superoxide Production in Trichinella Spiralis Infection.

The effect of probiotic strains (Enterococcus faecium EF55, E. faecium CCM7420, E. faecium CCM8558, E. durans ED26E/7, Lactobacillus fermentum CCM7421, L. plantarum 17L/1) on the production of superoxide anion (O2 -) in peritoneal macrophages of Trichinella spiralis infected mice was examined. E. faecium EF55 and E. faecium CCM8558 strains increased the O2 -production prior to parasitic infection,at the day7of application.A significant inhibition of the O2 - production caused by T. spiralis infection on day 5 post infection (p.i.) was prevented by all examined strains. Lactobacilli stimulated metabolic activity of macrophages during intestinal and early muscular phase (from day 5 to 25 p.i.) of trichinellosis. Enterococci increased the O2 - production in early intestinal phase (day 5 p.i.) and during the muscular phase of trichinellosis (days 25 and 32 p.i.). Respected increase in macrophage's metabolic activity induced by probiotic treatment in the intestinal phase of trichinellosis augmented the host antiparasite defence (damage and killing of newborn larvae with reactive oxygen species from macrophages).

The Anti-parasitic Effect of Probiotic Bacteria via Limiting the Fecundity of Trichinella Spiralis Female Adults.

A potential protective effect of probiotic strains against zoonotic Trichinella spiralis infection was investigated in the framework of a new therapeutic strategy aimed at using probiotics to control parasitic zoonoses. The study was focused on the impact of six selected probiotic (bacteriocinogenic) strains on the intensity of T. spiralis infection and female fecundity ex vivo and in vitro. Bacterial strains of different origin (Enterococcus faecium EF55, Enterococcus faecium 2019 = CCM7420, Enterococcus faecium AL41 = CCM8558, Enterococcus durans ED26E/7, Lactobacillus fermentum AD1 = CCM7421, Lactobacillus plantarum 17L/1) were administered daily in a dose of 109 CFU/ml in 100 µl, and mice were infected with 400 T. spiralis larvae on day 7 of treatment. Female adults of T. spiralis were isolated on day 5 post infection (p.i.) and subsequently were used in fecundity test ex vivo. E. faecium CCM8558, E. faecium CCM7420 and E. durans ED26E/7 strains significantly reduced the number of adults in the intestine. The application of L. fermentum CCM7421, L. plantarum 17L/1, E. faecium CCM8558 and E. durans ED26E/7 caused a significant decrease in the number of muscle larvae. The treatment with E. faecium CCM8558 and E. durans ED26E/7 showed the highest inhibitory effect on female fecundity (94 %). The number of newborn larvae (NBL) was also significantly decreased after administration of L. fermentum CCM7421 and L. plantarum 17L/1 (80 %). A direct impact of probiotic strains on female reproductive capacity was examined in vitro in females isolated from untreated infected mice on day 5 p.i. A correlation was found between the inhibitory effect and the concentration of probiotic strains. The reduction effects of the strains manifested as follows: L. fermentum CCM7421 (93 %), E. faecium CCM8558, L. plantarum 17L/1, E. faecium EF55 (about 80 %), E. faecium CCM7420 and E. durans ED26E/7 (about 60 %).

Small mammals: paratenic hosts for species of Toxocara in eastern Slovakia.

Toxocara spp., an aetiological agent of a serious helminthozoonosis, is a common roundworm of domestic and wild carnivores worldwide. The study aimed to estimate the seroprevalence of Toxocara in small mammals from different localities in eastern Slovakia. Anti-Toxocara antibodies were detected in 6.4% out of 2140 examined animals trapped in eastern Slovakia. Due to their high density and observed high seroprevalence of toxocariasis, Apodemus agrarius, A. flavicollis, Myodes glareolus and Mus spicilegus (10.9, 4.2, 3.6 and 11.2%, respectively) represent important sources of the infection. A significant correlation between type of food and Toxocara positivity was detected: granivores (7.2%) and invertebratophages (7.1%) were positive more frequently than herbivores (2.1%). In the years monitored, cyclic changes of seroprevalence were observed. A higher prevalence of antibodies in the spring was followed by a decrease in summer. In autumn, seroprevalence started to rise and stayed at a similar level through the winter. Seroprevalence of the examined animals confirms their contact with Toxocara spp. and demonstrates the presence of the aetiological agent in the monitored locality. Areas with a high prevalence of infected animals present constant infectious pressure on definitive hosts, thus also increasing infection risk for humans and paratenic hosts. The study confirmed the contact of small mammals with Toxocara spp. and demonstrated the presence and circulation of an aetiological agent in the localities monitored in eastern Slovakia.

Anti-tumour necrosis factor-alpha activity in Ixodes ricinus saliva.

Tumour necrosis factor-alpha (TNF-alpha) is one of the most prominent inflammatory mediators playing a central role in starting off the inflammatory reactions of the innate immune system. We identified a TNF-alpha-inhibitory activity in the saliva and salivary gland extract (SGE) from partially fed Ixodes ricinus ticks. Using mouse and human TNF-alpha specific ELISA, we showed that tick saliva or SGE markedly reduced the level of detectable cytokine. Both saliva and SGE inhibited the cytotoxic effect of TNF-alpha in a bioassay. Elimination of the TNF-alpha-inhibitory activity in SGE by trypsin digestion demonstrated that the anti-TNF-alpha factor is a protein. Fast protein liquid chromatography fractionation of SGE showed one peak of TNF-alpha-inhibitory activity corresponding to a protein with estimated molecular mass 23 kDa. The likely mechanism of the inhibitory effect is a direct binding of the cytokine. The TNF-alpha-inhibitory molecule seems to play an important role in the anti-inflammatory effect of tick saliva at the tick feeding site, providing a gateway to the host for tick-borne pathogens.

Immunomodulative effect of muramyldipeptide in mice with larval toxocarosis.

The phagocytic ability of polymorphonuclear leukocytes, the metabolic activity of peritoneal macrophages, the proliferative response of T- and B-cells, and the production of specific anti-Toxocara antibodies were studied in paratenic hosts with experimental larval toxocarosis after treatment with the immunomodulator muramyldipeptide for 119 days. Peroral infection of mice with 2,500 Toxocara canis eggs partially reduced the numbers of cells and inhibited the activity of all cellular immune-response parameters studied. During most of the experiment the greatest suppression was recorded for the peritoneal-macrophage metabolic activity. Parenteral administration of muramyldipeptide to mice at two doses prior to and after infection restored and significantly stimulated the parasite-inhibited phagocytic ability of blood polymorphonuclear leukocytes, metabolic activity of macrophages, and T-lymphocyte proliferative response. It also elevated the production of specific circulating anti-Toxocara antibodies. The immunomodulator significantly reduced the count of migrating T. canis larvae in the host organism (30.6%) and decreased by half the percentage of larvae in the brain.

The effect of cadmium on the immune behaviour of guinea pigs with experimental ascariasis.

The subchronic effect of cadmium on selected immunological parameters was studied in guinea pigs with experimental ascariasis. Cadmium chloride given orally for 28 days caused a considerable suppression of T- and B-cells in the lymphoid organs of intoxicated animals, of the metabolic activity of their peritoneal macrophages and a moderate decline in the level of complement CH50 and AH50 from day 1 to day 28 of the experiment, in comparison with the initial value. After a subsequent infection of the subchronically intoxicated guinea pigs the values for both the cell populations and the macrophage metabolic activity remained considerably suppressed, compared with infected control animals. The phagocytic and metabolic activity of macrophages as well as the CH50 and AH50 level, however, increased conspicuously for a short time after application of heavy metal and after infection but it did not reach the level of the infected group. The level of specific circulating antibodies was not affected by intoxication. Compared with the controls, the mean intensity of infection with Ascaris larvae migrating in the lungs of intoxicated animals increased by 20%. The results have established the negative effect of cadmium on the immunological parameters of the cellular immunity and humoral immunity studied and the impairment of the organism's response to the antigenic stimulus after an A. suum infection.

Effects of concurrently administered copper and mercury on phagocytic cell activity and antibody levels in guinea pigs with experimental ascariasis.

The subchronic effect of copper and mercury on selected immunological parameters in guinea pigs with experimental ascariasis was studied. Cupric sulphate and mercuric chloride administered for 28 days did not suppress the levels of specific circulating antibody in a subsequent Ascaris suum infection. Intoxication of animals significantly inhibited the phagocytic activity of peritoneal macrophages throughout the experiment. Ascaris suum infection elevated macrophage phagocytic activity but values remained significantly lower in comparison with infected non-intoxicated animals. Both the concurrently administered metal compounds changed the phagocytic ability of polymorphonuclear leucocytes in the blood. The intensity of infection assessed by the number of Ascaris larvae migrating in the lungs of animals currently intoxicated with both the metal compounds increased by 28.6%, compared with controls.