Herpesvirus diseases of domestic animals and game species in the Slovak Republic.

Herpesviruses are DNA viruses that cause serious latent infections in humans and animals. These pathogens significantly influence the animal health and economy of animal husbandry. The reduction of production parameters, abortions, birth of weak individuals and by costs associated with the elimination and monitoring of herpesvirus diseases are among the most serious harms caused by herpesviruses. In our review we focused mainly on herpesvirus diseases in pigs and cattle (Aujeszky's disease, bovine infectious pustular vulvovaginitis and balanoposthitis, infectious bovine rhinotracheitis) and herpesvirus diseases in red deer, dogs, and carps. In the past, these herpesvirus diseases have caused considerable economic losses in livestock. At present, several of these diseases are eliminated in the Slovak territory. Currently, a continuous monitoring is carried out not only in populations of domestic animals, but also in wild animals, which are the main reservoirs of the mentioned herpesviruses. Keywords: herpesvirus; Aujeszky's disease; animals; fish; eradication; Slovak Republic.

Detection of Mycobacterium avium subsp. paratuberculosis in Slovakian wildlife.

Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of infectious enteritis called paratuberculosis that has a high economic impact on the worldwide livestock production. A central important question arises: Can wildlife animals serve as a reservoir for transmission of MAP to domestic ruminants? With this in mind, we devised a study to detect MAP in various Slovakian wildlife species found in the areas where paratuberculosis had been documented in domestic ruminants. The samples of parenchymatous organs (intestines, ileocecal valve and mesenteric lymphatic nodes) from 83 wildlife animals representing 13 species, inclu- ding 7 herbivorous, 5 carnivorous and 1 omnivorous species were collected during a four-year period. The clinical and pathological examinations failed to demonstrate any manifestations of paratuberculosis in any of the wildlife samples. The detection of MAP was done by widely used tests, i.e. cultivation and the PCR analysis. The bacterial cultures revealed the growth of Mycobacterium spp. colonies in 58 (70%) of all of the wild animals, but the PCR testing demonstrated paratuberculosis only in one (7.69%) of the roe deer population.

The roundworm Strongyloides stercoralis in children, dogs, and soil inside and outside a segregated settlement in Eastern Slovakia: frequent but hardly detectable parasite.

A comparative study was carried out to evaluate the Strongyloides stercoralis infections in children and dogs inside and outside the segregated settlement in Medzev, Eastern Slovakia, and a survey of the soil within the settlement was included. Applying the Koga agar plate (KAP) culture method and microscopy examination of stool samples collected from 60 Roma and 21 nonRoma children, no larvae of S. stercoralis were detected but eggs of three nematodes (Ascaris lumbricoides, Trichuris trichiura, and Enterobius vermicularis) and cysts of two protozoan endoparasites (Giardia duodenalis and Cryptosporidium spp.) were often found. However, immunoenzymatic assay (ELISA) for the evidence of IgG antibodies against S. stercoralis showed 33.3% seroprevalence in Roma children and 23.8% prevalence in children from the majority population, attending the same school. Eosinophilia was regularly present in children with exclusive infection of S. stercoralis (eight cases) as well as in individuals suffering from mixed infections of S. stercoralis and some of the above listed parasites (16 cases); high eosinophil counts sometimes, but not always, occurred in parasitized children lacking S. stercoralis antibodies. A comparison of S. stercoralis in dogs from the settlement (40 dogs) and from a distant dog shelter (20 dogs) did not reveal remarkable differences: the direct microscopy of faecal samples revealed rhabditiform larvae in 13.3% of the dogs from the settlement (4/30) and in 10.0% of the dogs from the shelter (2/20). Out of blood samples collected from the second dog group, 55% of the dogs contained antibodies against S. stercoralis. In the soil collected from 14 various locations within the settlement, S. stercoralis larvae were observed in two samples (14.3%); however, 13 samples (92.9%) were positive for human or dog endoparasites of the genera Ancylostoma, Ascaris, Toxocara, Toxascaris, Trichuris, and Hymenolepis.

Giardia duodenalis and Giardia enterica in children: first evidence of assemblages A and B in Eastern Slovakia.

For an overview on the occurrence of Giardia assemblages in children in Eastern Slovakia, we examined 259 faecal samples of children from the segregated settlement in Medzev, 30 samples of children from the orphanage in Medzev and 40 samples of children with autism from the Special Elementary School in Kosice. Thirty-eight samples (14.67 %) from the segregated settlement, 19 samples (63.33 %) from the orphanage and two samples (5.0 %) from the Special Elementary School were positive for Giardia by flotation. The initial microscopic diagnostics were completed by the genotyping of the triosephosphate isomerase-gene loci (tpi genes) which revealed the existence of two Giardia assemblages in Slovak population, namely Giardia duodenalis (assemblage A) and Giardia enterica (assemblage B). These results represent the first evidence of A and B assemblages in children in Slovakia. Epidemiological significance and the impact on the public health of Giardia infection are highlighted.

[Immunomodulatory effect of levamisole and administration of amitraz in dogs with uncomplicated generalized demodicosis]. / Imunomodulacný efekt levamizolu pri pouzití amitrazu u psov s nekomplikovanou generalizovanou demodikózou.

The immunomodulatory effect of levamisole (Decaris tbl.) in the course of acaricide therapy with amitraz (Taktic) on the functional activity of blood neutrophils (% of phagocytizing cells and ingestion capacity) and lymphocytes (blastogenic response to Con A) in dogs with uncomplicated generalized demodicosis (NGD) was studied. The level of examined parameters was evaluated before treatment, week 3 and 7 after the first application of these preparations; and compared with the values of NGD dogs treated only with amitraz and with those in clinically healthy dogs. In comparison with healthy dogs the initial level of examined activities of both cell populations was significantly depressed. A significantly earlier (4 weeks earlier) increase (when compared with values before treatment) of investigated activities of neutrophils and lymphocytes occurred in dogs treated with amitraz and levamisole in comparison with those in dogs treated only with amitraz. It was manifested especially significantly in phagocytosis, the ingestion capacity of neutrophils at this time of therapy has reached the level of those in healthy dogs. Functional activity of lymphocytes in both groups of NGD dogs has not reached a comparable value with that in healthy dogs either at the end of observation. The presented results indicate that significantly earlier improvement of functional activity of phagocytes and lymphocytes in demodectic dogs treated with amitraz and levamisole was connected with the immunorestorative effect of levamisole.

Lymphocyte blastogenesis to concanavalin A in dogs with localized demodicosis according to duration of clinical disease.

The aim of the study was to determine the degree of Con A induced lymphocyte blastogenesis in dogs with localized demodicosis (LD) within 1-3 and 6-8 weeks from appearance of the clinical signs. Ethidium bromide fluorescence assay was used for evaluation. In observation 9 clinically normal dogs, 6 dogs with LD a 4 dogs with generalized demodicosis (GD) were used. The results showed a statistically significant depression (P < 0.01) of blastogenesis in the LD dogs in comparison with that in the healthy dogs (Fig. 1). Responses to Con A were normal in dogs with LD in 1-3 weeks. However, a significantly depressed response to Con A (P < 0.025-0.001) was demonstrated in the LD dogs in 6-8 weeks (average 6.7 weeks) and it was comparable with that in the GD dogs with the duration of clinical disease on average for 8.7 weeks (Fig. 2; Tab. II). Thus, immunosuppression is not a necessary condition for dogs to develop spontaneous clinical LD and immunosuppression develops with the clinical signs of disease.

The immunomodulatory effect of the soluble fungal glucan (Pleurotus ostreatus) on delayed hypersensitivity and phagocytic ability of blood leucocytes in mice.

The effect of fungal and yeast glucan on different immune functions in mice was examined and compared. The simultaneous administration of glucan and a sensitizing dose of DNFB on the different sites significantly stimulated delayed-type hypersensitivity (DTH) response only when using fungal glucan. Both glucans tested, when administered before sensitization, significantly increased DTH response, but with a significantly higher level at the beginning of the investigation (on day 7) when using fungal glucan. The increase in phagocytic activity by the blood leucocytes started in the 1st week after fungal-glucan treatment, and in the 2nd week after yeast-glucan treatment, and took longer after administration of fungal glucan. The values of the phagocytic-activity index were significantly influenced only after fungal-glucan injection. The results of the study indicate that fungal glucan isolated from Pleurotus ostreatus could be a prospective immunomodulating substance.

Evaluation of canine lymphocyte blastogenesis prior and after in vitro suppression by dog demodicosis serum using ethidium bromide fluorescence assay.

The mitogen induced blastogenic response of lymphocytes from normal dogs and dogs with generalized demodicosis (GD) was measured by the ethidium bromide (EB) fluorescence assay. Serum from GD dogs significantly suppressed the in vitro reactivity to Con A of peripheral blood lymphocytes (PBL) from normal dogs and GD dogs, however with a different percentage of suppression 40.6 and 81.2%, respectively. As a result, the degree of lymphocyte blastogenesis suppression in GD dogs did not parallel the immunosuppressive potency of their serum (Tab. IV). The data indicate that PBL obtained from GD dogs did not respond to Con A as well in the presence of serum from normal dogs as did PBL from normal dogs (Tab. IV). In one third of examined GD dogs a similar situation was described also by Hirsh et al. (1975). The basis for this cellular modified response is unknown. It does not appear that the age or the chronicity of the disease are related to this observation. Further studies are necessary to elucidate this relation. The GD dogs showed not only a significant depression of the lymphocyte response to Con A but also enhancement of the ability of unstimulated cells to proliferate was also observed (Tab. IV). Similar observation was reported by others (Barriga et al., 1992). The meaning of this is not clear at present. This finding is discussed in the light of proposed different effects of the parasite or the host's reactivity to the parasite on different subsets of lymphocytes. No significant difference of PBL responsiveness to Con A between healthy dogs with respect to the age (Tab. III) and the time of examination (compare results in Tabs. I and IV) was observed. Autologous serum showed a better responsiveness of normal canine lymphocytes to Con A than fetal calf serum (FCS). It is suggested that the use of FCS might lead to an erroneous judgement (Tab. I). Both lectins, Con A and PHA induced cell proliferation of healthy dogs in very similar amount (Tab. II). Our results indicated that EB fluorescence assay is a useful method for detection a respondence of canine lymphocyte blastogenesis.

[Evaluation of indicators of cellular immunity in experimental infectious bovine rhinotracheitis virus infection in calves treated with glucan]. / Hodnotenie niektorých ukazovatel'ov bunecnej imunity po experimentálnej infekcii vírusom IBR u teliat osetrených glukánom.

Some parameters of cell-mediated immunity (phagocytic activity PA and phagocytic index PI, metabolic activity and migration-inhibitive factor MIF of blood leucocytes) were investigated in calves experimentally infected with IBR virus (group II) and in calves premedicated with glucan seven days before infection (group I). A significant increase in the percentage of phagocytizing cells was observed in both groups of animals on days 2-4 of the experiment (P < 0.01) in comparison with the values before infection. If the two groups of animals were confronted with each other, there was a significant difference in the percentage of phagocytizing cells on days 1 and 2 after infection (P < 0.05), with the higher average values in the calves which were only infected. An evaluation of PI showed a statistically significant difference only in the glucan-treated calves on day 7 after its administration (P < 0.05), i.e. on day zero of the experiment. The inhibition of leucocyte migration ability was found on day 1 after infection, and it persisted for four days, without any significant differences between the groups. An evaluation of the metabolic activity of leucocytes (INT-test) showed a decrease in the calves of both groups in comparison with the initial value, without any significant differences when the values were confronted with each other.

[The effect of an insoluble fungal glucan on indicators of immunity in calves]. / Vplyv nerozpustného fungálneho glukánu na niektoré ukazovatele imunity teliat.

Immunomodulative effect of insoluble fungous glucan (IFG) was observed on nine accidentally chosen calves, which were in the age from 4.5-5.5 months. The following items were observed: T, B lymphocytes, white blood cell count, index and activity of phagocytosis, induction and determination of interferon from leukocytes. In the experimental group there was a decrease in B-rosseting cells observed. On the 28-th day and on the 42-th day of experiment duration a significant increase in B-rosseting cells in comparison with control group has been observed. Average values were significantly different on the levels P < 0.01 and P < 0.05. Percentages of T-rosseting cells were almost the same during the whole experiment. In the experimental group on the 7-th and on the 14-th day the decrease has been observed. By the Student's t-test a significant difference in favour of group, where IFG was applicated, has been observed. Percentage of white blood cell count was from 47 to 72 in both groups. When phagocytic activity was evaluated, there was the higher percentage of phagocytic cells in the 7th, 14-th, 21-st, 28-th and 42-th day of experiment when compared with the 0-day value. Concerning the phagocyte index a statistical difference was observed, when compared to the initial value in favour of the experimental group. The initial disbalanced values of interferon, which have been observed during the period from the 7-th to the 21-st day, increased on the 28-th and the 42-nd day of experiment which was statistically confirmed on the following levels: P < 0.05 and P < 0.01.

[The effect of fungal and yeast glucan and levamisole on the level of the cellular immune response in vivo and leukocyte phagocytic activity in mice]. / Vplyv fungálneho a kvasinkového glukánu a levamizolu na úroven celulárnej imunitnej odpovede in vivo a fagocytárnu aktivitu krvných leukocytov u mysí.

The level of cell-mediated immune response in vivo was investigated using the test of delayed hypersensitive reaction (DHR) to DNFB, along with the phagocytic activity (PA) of blood leucocytes in mice after subcutaneous implantation of fungal and yeast glucan and levamisol in dependence on the dose and administration schedule. The soluble form of fungal glucan (Pleurotus ostreatus) potentiated the DHR significantly at a dose of 10 mg/kg (but not at a dose of 50 mg/kg) while it was administered during DNFB sensitization (P < 0.05)-Tab. I and when its pre-medication effect was investigated (days -7 and -14; P < 0.05) with regard to the time of sensitization (Tab. II). The identical dose of glucan also had a positive effect (P < 0.05 or 0.01) on the percentual proportion of phagocytic cells (PC) reaching the maximum in the 2nd and 3rd week of investigation, as well as on the phagocytic activity index (P < 0.05; 3rd week) and percentage of neutrophil granulocytes (P < 0.05; 2nd week)-Tab. III. Yeast glucan (Saccharomyces cerevisiae) showed a potentiating effect on the DHR to DNFB only in the case of its pre-medication use; its soluble form was effective at both doses (10 mg and 50 mg/kg) in days -7 and -14 (P < 0.05), and its corpuscular form at a dose of 50 mg/kg on days -7, -14 and -21 (P < 0.05 or 0.01)-Tab. II. PA parameters of blood leucocytes displayed a stimulative effect only on the PC percentage. The most significant effects in this case were observed in the soluble form (both doses) in the 2nd and 3rd week (P < 0.01 and 0.05, resp.) and in the insoluble form (both doses) in the 3rd and 4th week of observation (P < 0.05 and 0.01, resp.). An increase in the number of neutrophil granulocytes was significant in the 2nd (P < 0.05 or 0.01; corpuscular form) and 3rd week of the experiment (P < 0.01; soluble form)-Tab. III. Levamisol affected both investigated parameters (DHR and PA) only at a dose 20 mg/kg (10 mg/kg-no effect). Its potentiating effect on the DHR level was observed both for its administration at the time of sensitization (P < 0.05) and for its administration on days 7 (P < 0.05) and 14 (P < 0.01) before DNFB sensitization (Tabs. I and II). A statistically significant increase in PC was recorded in weeks 2, 3 and 4 (P < 0.05 or 0.01), a statistically significant increase in the number of neutrophil granulocytes in the 3rd week of investigation (P < 0.05). The phagocytic activity index was not affected.

[The effect of administration of non-soluble fungal glucan on the level of delayed skin hypersensitivity and phagocytic activity of leukocytes in the blood of calves]. / Vplyv aplikácie nerozpustného fugálneho glukánu na úroven oneskorenej koznej precitlivelosti a fagocytárnu aktivitu krvných leukocytov u teliat.

The level of delayed skin hypersensitivity (DSH) was evaluated in vivo after dinitrofluorobenzene induction (3rd and 7th week of the experimental) while the level of phagocytic activity (FA) of blood leucocytes (in weekly intervals during six weeks of observation) was evaluated in calves (4.5 to 5.5 months of age) after administration of particular fungal glucane (from oyster mushroom) at a dose of 10 mg/kg live weight. In calves which were administered glucane (abbr. G), a significantly stronger cell-mediated immune response was recorded by means of DSH test in the 3rd week of observation, namely in comparison with its starting value (P < 0.01) and also with the value of control animals (P < 0.01; K group)-Tab. I. In the seventh week of the experiment there was not a statistical difference in the average values of DSH any more (Tab. I). But at that time 100 percent of calves showed a skin reaction above 6.5 mm (mean = 7.64 +/- 0.55) evoked by the used sensibilizer in the G group; this reaction was significantly higher (P < 0.01) in comparison with the value of DSH in a majority (80%) of calves of the K group (mean = 6.15 +/- 0.21)-Fig.1. The immunostimulating effect of the used glucane on the FA of blood leucocytes was less significant.(ABSTRACT TRUNCATED AT 250 WORDS)