Experimental oral and ocular Encephalitozoon cuniculi infection in rabbits.

Encephalitozoon cuniculi is an obligate intracellular pathogen that has a wide host distribution, but primarily affects rabbits. The aim of this study was to characterize both the cell-mediated and the antibody response in rabbits after experimental infection using 2 different infection routes: oral and ocular. SPF rabbits were infected with low (10³ spores) and high (107 spores) infection doses. Monitored parameters included clinical signs, detection of spores in urine, antibody response detected with ELISA, and cell-mediated immunity detected by antigen-driven lymphocyte proliferation. At week 13 post-infection, half of the rabbits in each group were suppressed by intramuscular administration of dexamethasone. At week 18 post-infection, animals were euthanized. Clinical signs were mild with exacerbation after immunosuppression. Spores in urine and antigen-specific cell-mediated immunity were detected from weeks 5 and 4 post-infection, respectively. Specific IgM was detected 1 week after infection, and IgG antibodies followed 1 week later in rabbits infected with the high dose. Immunological responses were dose dependent. The authors can conclude that both oral and ocular experimental infection with E. cuniculi resulted in an immune response of the infected animals. Rabbits could be used as an experimental model for the study of ocular microsporidiosis.

Involvement of the protein tyrosine phosphatase SHP-1 in Ras-mediated activation of the mitogen-activated protein kinase pathway.

Ubiquitously expressed SH2-containing tyrosine phosphatases interact physically with tyrosine kinase receptors or their substrates and relay positive mitogenic signals via the activation of the Ras-mitogen-activated protein kinase (MAPK) pathway. Conversely, the structurally related phosphatase SHP-1 is predominantly expressed in hemopoietic cells and becomes tyrosine phosphorylated upon colony-stimulating factor 1 treatment of macrophages without associating with the colony-stimulating factor 1 receptor tyrosine kinase. Mice lacking functional SHP-1 (me/me and me(v)/me(v)) develop systemic autoimmune disease with accumulation of macrophages, suggesting that SHP-1 may be a negative regulator of hemopoietic cell growth. By using macrophages expressing dominant negative Ras and the me(v)/me(v) mouse mutant, we show that SHP-1 is activated in the course of mitogenic signal transduction in a Ras-dependent manner and that its activity is necessary for the Ras-dependent activation of the MAPK pathway but not of the Raf-1 kinase. Consistent with a role for SHP-1 as an intermediate between Ras and the MEK-MAPK pathway, Ras-independent activation of the latter kinases by bacterial lipopolysaccharide occurred normally in me(v)/me(v) cells. Our results sharply accentuate the diversity of signal transduction in mammalian cells, in which the same signaling intermediates can be rearranged to form different pathways.

Commercially available rabbit anti-human polyclonal antisera as a useful tool for immune system studies in veterinary species.

We have used selected rabbit anti-human polyclonal antibodies as an example of useful and easily available tools for studies on immune system structure and development in important veterinary species, many of which also represent animal models in biomedicine. The cocktail of anti-human Igkappa-FITC/anti-Iglambda-RPE F(ab')(2) fragments was used for two-colour and, in combination with the cross-reactive anti-CD79alpha monoclonal antibody HM-57, for three-colour flow cytometry of canine, feline, bovine and porcine peripheral B-cells. A possible application of such immunoreagents in studies on primary B-cell differentiation has been suggested in pigs; the same approach can be used in other species of interest. Rabbit anti-human lactoferrin-FITC F(ab')(2) fragment was used for visualizing neutrophils in dogs, pigs and cattle and an application for two-colour immunophenotyping of canine granulocyte subsets has been designed. Affinity isolated rabbit anti-human CD3 and anti-human TdT have been shown to represent a ready-to-use tool for in situ studies on primary T-lymphopoiesis in pigs with possible extensions both to the B-lineage development in pigs and other animal models. Altogether, our study show that carefully selected polyclonal antibodies available on the market may possess broad cross-reactivity with important applications in veterinary research.

Superovulation in cattle: Effect of goat anti-PMSG serum.

Sixteen heifers were superovulated using 5 000 i.u. PMSG on days 9 to 11 of the oestrous cycle (day 1 of the experiment) followed by two injections of 500 mug Estrumate 48 and 54 hours later. Eight of them were injected with goat anti-PMSG serum 5 hours after the first signs of oestrus were observed. Compared with the control group, the treatment with anti-PMSG serum resulted in a shorter heat period (25.8 vs. 51.3 hours), a higher mean number of ovulations (22.1 vs. 18.0) and a lower number of follicles over 10 mm in diameter (4.1 vs. 22.3). The mean numbers of eggs recovered in the experimental and control groups were 17.8 and 6.9, of which 70.2 and 42.0 per cent, respectively, were viable embryos. The concentrations of progesterone and 17-beta oestradiol in the blood plasma showed no significant differences between the experimental and control animals. A higher oestradiol in the control group on day 9 of the experiment was in keeping with the histological picture of the target organs and with a significantly higher number of follicles at slaughter on days 12 to 14 of the experiment.

Regulation of bovine labor with a long-acting carba-analog of oxytocin: a preliminary report.

The long-acting oxytocin (OT) analog 1-desamino-1-monocarba-E12-Tyr(OMe)]-OT(dCOMOT) was given IV to 13 pregnant cows near to term, but not in actual labor. The animals were para 1 to 5. Of these cows, four were treated with 20 mg of dexamethoasone 48 hours before the peptide was injected; the remaining nine animals were given no other medication. The animals usually were given a single injection of 5 mg of dCOMOT into the jugular veiw (5 to 7 micrograms/kg of body weight). In those instances where delivery was not complete within six hours, a second injection of peptide was given. In all instances, the first injection initiated labor as judged by the behavior of both uterus and cow. In most instances, there was also intermittent spurting of colostrum from the udder over a six-hour period after injection. In the four cows treated first with dexamethasone, the mean duration of induced labor was 4.35 hours after dCOMOT injection. In the nine non-treated cows, the mean duration of induced labor was 14.25 hours. The difference between the two values was significant. For both groups separately, and all data together, there was a linear inverse relation between the size of the external ostium uteri at injection and the duration of labor after peptide injection. All calves were healthy with no signs of hypoxia and the dose rates used did not result in any instance of uterine tetany or tachyphylaxis.

[Enzymatic activity in the placenta in cows in relation to the course of parturition]. / Enzymatická aktivita v placentomech krav ve vztahu k prûbehu porodu.

The activity of aminopeptidases and cathepsins was determined in placentoma homogenates; placentomas of cows were extirpated immediately after parturition and in four and eight hours. In cows with afterbirth retention (a. r.) following induced parturition, the activity of these enzymes was always higher than in cows without a. r., no matter if after induced and spontaneous parturitions; it was at a similar or slightly higher level than in the eighth month of pregnancy. The content of total proteins in placentomas of cows with a. r. was also higher. These findings point to the insufficient ripening, or aging of placental tissue, which is related to a release of lysosomal enzymes. The enzymes are active through hydrolysis in the separation of foetal placenta from the maternal one and in the modification of proteins participating in intercellular linkages.

[Activity of the ovarian follicle system in cows with various puerperal courses]. / Aktivita folikulárního systému ovarií krav pri rozdílném prubehu puerperia.

The presence of the various categories of follicles (less than 0.05; 0.5-1.0; 1.0-1.5; less than 1.5 cm) in the ovaries was examined by palpation, endoscopically and post mortem in cows with a physiological puerperium (n = 5), with puerperal endometritis (n = 5), and with retention of afterbirth (RS, n = 5) from the second to the 20th day post partum (p.p.). The largest proportion (82.6%) of the smallest follicles (less than 0.5 cm) was recorded between the fourth and seventh day p.p. in clinically healthy cows; a marked decrease was then observed until the 15th day p.p. In cows with endometritis the proportion of the smallest follicles was only 60%. The average number of follicles 0.5 to 1.0 cm in size increased 1.6 times in the healthy cows between the 7th and 15th day p.p., but in cows with pathological puerperium their number declined. The largest follicles (greater than 1.5 cm) occurred in the cows with pathological puerperium as late as at the end of the period of study, whereas in healthy cows their occurrence was fairly frequent sooner--between the 8th and 11th day p.p. In all groups of cows the average number of follicles of all size groups declined until the 12th day p.p. which is an indication of the finished first growth wave. Occurrence of the first follicle larger than 1 cm in diameter was observed in 73.3% of cases in the contralateral ovary, opposite to the originally gravid uterine horn. It is assumed that delayed and asynchronous growth of follicles in cows with pathological puerperium is a consequence of disturbed repairing processes of endometrium and of endocrine dysfunction.

[Histology of the endometrium in cows in normal and pathologic puerperium]. / Histologie endometria krav v prubehu fyziologického a patologického puerperia.

There were repeated histological bioptic examinations of the endometrium during the period from the second to twentieth day post partum (p.p.) in cows with afterbirth retention (n = 5), with developing puerperal endometritis (n = 5), and with a physiological course of puerperium (n = 5). In cows with a physiological course of puerperium, a finished process of epithelization of endometrium with a morphological and functional consolidation of the epithelium could be observed on the 20th day p.p. In cows with disturbed puerperium, the surface epithelium still exhibited signs of disintegration and a low secretion activity. The endometria of both groups of cows with pathological puerperium had a high degree of infiltration with neutrophil granulocytes: in cows with retained afterbirth this picture was characteristic already the second day p.p. On the other hand, the endometrium of cows with physiological puerperium was characterized by a higher occurrence of folliculoid nodules as early as on the tenth day p.p., and a rapid increase in the number of fat cells and activated histiocytes--macrophages could be observed in the endometrium of these cows between the 15th and 20th day p.p. The dynamics of occurrence of mobile cells in the studied groups of cows confirmed their importance for a physiological course of the restitution of endometrium during the puerperium of cows.

[The effect of repeated administration of Oestrophan (SPOFA) on the quality of sexual function in breeding stock]. / Stanovení vlivu opakovaného podání Oestrophanu (SPOFA) na kvalitu pohlavních funkcí plemenic.

The objective of the study was to check a contingent negative effect of repeated treatment of heifers by PGF2 alpha--Oestrophan (SPOFA)--on the course and quality of their ovarian activity and external heat signs. Changes in progesterone concentrations were evaluated three times a week. Three heifers were subjected to treatment eight times and two heifers nine times. Of 42 applications of Oestrophan (SPOFA) the response was positive in 95.2% (forty heats). When Oestrophan (SPOFA) was administered on the fourth and sixth day of the sexual cycle, in two cases no heat nor luteolysis of corpus luteum was detected; this was demonstrated by hormonal investigation. Of the forty evaluated heats, 82.5% were classified as strongly and medium expressive, the remaining were weakly expressive. The onset of heat was found to be on the average 72.6 hours after Oestrophan (SPOFA) administration, in 9.1% of the cases the heat started about 48 hours after administration, 12.4% of cows showed the heat signs between 72nd and 96th hours.

[Histologic and histochemical study of placentomes in cows after induced labor]. / Histologické a histochemické vysetrení placentomu krav pri indukovaných porodech.

The placentomes were extirpated from 16 cows after parturition induced with 750 micrograms cloprostenol or 20 mg dexamethasone on the 277th day of gravidity, on an average, from 9 cows after spontaneous parturition, and from 7 cows after hysterectomy in the eighth month of gravidity. In the cows with induced calving the foetal placenta was not expelled within 12 hours after calving in 68.7% of the cases whereas in the spontaneous parturitions this proportion was only 22.2% of cases. The placentomes obtained immediately after calf expulsion, and then after four and eight hours, were subjected to histological and histochemical examination. In the terminal crypts of the placentome in cross sections obtained from cows which expelled the placenta in time after natural and induced parturitions, the number of binuclear cells of the fetal syncytium and of cells of the dam epithelium (P less than 0.001) was found to be significantly lower than in the cases of afterbirth retention (1.2 and 3.9; 6.4 and 18.5). The cells of the cow's epithelium of the expelled placentae had a higher activity of acid phosphatase and lipids and the foetal syncytium had a higher activity of non-specific esterase. Increased alkaline phosphatase activity was characteristic of the cow's epithelium in the cases of subsequent retention of afterbirth. These findings should be taken into account in efforts for developing new methods of the induction of parturition if the undesired occurrence of afterbirth retention is to be reduced.

Light and scanning electron microscopy of endometrium of ovariectomized cows treated with oestradiol.

Histomorphological and histochemical investigations showed that a single intramuscular dose of 3 mg or oral dose of 10 mg oestradiol were sufficient to induce physiological oestrogenization of endometrium in ovariectomized cows. The first signs of endometrial activation were observed already on post-treatment Day 2. The resumption of secretory activity of surface epithelium cells and epithelial cells of proximal segments of uterine glands, active hyperaemia, slight oedema of lamina propria, and infiltration with cells responsible for non-specific resistance of endometrium (lymphocytes, lymphoid cells, monocytes, macrophages, mast cells) were the typical signs of oestrogenic stimulation. Morphological and activation signs of the effects of oestradiol persisted up to post-treatment Day 9. High doses of oestradiol (10 mg intramuscularly or intravenously, 20 mg orally) induced non-physiological hyperoestrogenization of uterine mucosa.

[The effect of estrogens on phagocytic activity of blood leukocytes in heifers and cows]. / Vliv estrogenu na fagocytární aktivitu krevních leukocytu jalovic a krav.

Phagocytic activity of peripheral leucocytes (PA) was measured in heifers during the luteal phase and oestrus and after administration of various doses of oestradiol, in ovariectomized heifers and cows in the early post-partum period. PA was demonstrated in 22.63 +/- 2.49% and 50.61 +/- 3.76% of phagocytes in the luteal phase and oestrus respectively (P less than 0.01, Fig. 1). The phagocytic index (PI) rose parallelly from 2.26 +/- 0.31 to 6.55 +/- 0.64 particles per cell (Fig. 2). Intramuscular administration of a single dose 3 mg of oestradiol dipropionate resulted in an increase of PA from 28.93 +/- 3.34 to 69.60 +/- 3.32 on post-treatment day 3 (P less than 0.05, Fig. 3). A nonsignificant increase of PA was observed in heifers treated with 10 mg oestradiol. Increases of PA and PI in postparturient cows, treated with various doses of oestradiol, were nonsignificant (Fig. 5, 6) owing to a wide variance of the values obtained, which might be due to individual differences in endocrine and metabolic status or to uterine bacterial contamination. The most marked of haematological changes was the increase of eosinophil counts from 317 to 525.10(6).l-1 in a group of cows treated with 10 mg oestradiol and a decrease of the lymphocyte: neutrophil ratio (Tab. I). Both endogenous and exogenous oestrogens stimulate PA of peripheral leucocytes and 3 mg of oestradiol is a sufficient dose to obtain the effect.

Stimulation of cell defense mechanism of bovine endometrium by temporal colonization with selected strains of lactobacilli.

The aim of the experiments was to assess the response to topical administration of selected live strains of lactobacilli of the cells responsible for the resistance of bovine endometrium. Experimental cows (n = 8) at 8 to 12 days of the estrous received one intrauterine dose of 20 ml of a suspension of lactobacilli in 1% glucose solution. Group I (n = 4) was treated with the strain Lactobacillus spp. G 013 (5.5 x 10(8) CFU/ml) and Group II (n = 4) with the strain Lactobacillus casei CCM 1753 (1.2 x 10(8) CFU/ml). Control cows (n = 4) received 20 ml of 1% glucose solution. Samples of endometrial tissue were obtained by biopsy or from slaughtered cows on post-treatment days 5 or 6 and/or 10 or 12. Colonization of the uterine cavity with lactobacilli for up to 12 days was confirmed by bacteriological examination and scanning electron microscopy. Highly significant increases (P < 0.01) were found in numbers of all cell types under study. The pronounced cellular infiltration of endometrium was mostly due to the accumulation of mononuclear cells, particularly lymphocytes forming often indistinctly demarcated lymphoid nodules. Also marked was the infiltration of mast cells and macrophages. The cellular infiltration of endometrium persisted still on post-treatment day 12. No signs of alteration of epithelial cells were observed. No principal differences in the effects on endometrium were found between the two lactobacilli strains. The proved stimulatory effect of lactobacilli on endometrial cell defense mechanisms demonstrated in our experiments and inhibitory effects of the former on the growth of pathogenic microorganisms are promising for practical application in the prevention and alternative therapy of bovine endometritis.

[Levels of progesterone, 17 beta-estradiol and 11-hydroxycorticosteroid in the blood in cows in different types of puerperal conditions]. / Koncentrace progesteronu, 17 beta-estradiolu a 11-hydroxykortikosteroidü v krevní plazme krav pri rozdílném prubehu puerperia.

Changes in progesterone (P4), 17 beta-estradiol (E2) and 11-hydroxycorticosteroid (11-OHCS) concentrations in blood plasma were determined in daily intervals from the day of parturition to day 15 post partum (p.p.) in cows with physiological puerperium (n = 8), with puerperal endometritis (n = 6), and with placenta retention (n = 6). Cows with puerperium disorders (endometritis, placenta retention) had significantly higher P4 levels in the period from day 3 to day 7 p.p. than cows with physiological puerperium. E2 concentrations decreased to basal values following the parturition in cows with spontaneous parturitions and subsequent expulsion of the placenta. A delayed decrease in E2 concentrations after parturition and a significant increase on day 5 and day 7 p.p. were recorded in the group of animals with placenta retention. Significantly higher levels of 11-OHCS in blood plasma were detected by day 5 or by day 8 p.p. in cows with placenta retention and puerperal endometritis. Extraovarial sources of sexual steroids resulting in endocrine malfunctions are discussed as well as likely consequences for puerperium.

[Detection of Chlamydia in inflammation of the cervix and uterus in cows]. / Nálezy chlamydií u zánetu delozního krcku a pochvy krav.

Clinical findings in cows of seven farms with a frequent occurrence of birth canal inflammations were matched against results of serological and direct immunofluorescence tests for Chlamydia sp. infection. Normal clinical picture, weak to moderate and serious inflammations were found in 16.1%, 46.7% and 37.2% of the 205 examined cows, respectively. The results of micro CFT were negative in 32.7% and titres 1:5, 1:10, 1:20, and 1: > or = 40 were found in 8.7%, 38.0%, 12.3%, and 8.3%, respectively. Direct immunofluorescence test of cervical swabs, using the Progen Biotechnic GmbH set, was negative in 29.1% and clearly positive in 34.7% of the tested cows. The rest was animals in which sporadic elementary bodies were seen. The significance of the findings is discussed and the necessity to test Chlamydia infections in herds affected by frequent and/or chronic inflammations of genital organs is stressed. Evidence of the transmission of Chlamydia infection by contaminated semen is presented.

[Influencing superovulation in cattle with serum gonadotropin antibody]. / Ovlivnení superovalace u skotu protilátkou proti sérovému gonadotropinu.

The effectiveness of GnRH (Luliberin) and goat anti-PMSG serum prepared at the Veterinary Research Institute, Brno, was studied during the superovulation of heifers treated with PMSG and Estrumate. The use of GnRH did not exert any positive influence on superovulation, as compared with the control groups,. when anti-PMSG serum was administered five hours from the determined beginning of oestrus (46-53 hours from the first administration of Estrumate), the length of oestrus was reduced in comparison with the control group (25.8 and 51.3 h), the number of 22.3). The tested heifers produced 17.8 ova, on an average, out of which (4.1 and 22.3). The tested heifers produced 17.8 ova, on an average, out of which 12.5 were good-quality embryos. The control heifers produced 6.9 ova including 2.9 embryos. The obtained results provide convincing evidence of the promising results of the use of goat anti-PMSG serum during the superovulation cattle.

Passive immunisation of post-weaned piglets using hyperimmune serum against experimental Haemophilus parasuis infection.

The protective role of hyperimmune serum in the prevention of Haemophilus parasuis infections in post-weaned piglets was assessed by experimental challenge. The hyperimmune serum was obtained from a pig vaccinated with a commercial vaccine against Glässer's disease. Thirty-eight weaned piglets were divided into four groups: three groups were immunised intramuscularly with 10 ml of hyperimmune serum and one group consisted of unimmunised control animals. All piglets were subsequently infected intraperitoneally with H. parasuis serotype 5 at different times after immunisation. The use of hyperimmune serum provided the piglets with partial protection against experimental infection. The levels of protection indirectly depend on time between serum inoculation and challenge infection. The best protection of piglets against experimental infection was obtained in the group immunised 1 week before inoculation; the same group in which the highest levels of antibodies were detected at the time of challenge.

['Microbial air pollutants'. Emission sources and preventive measures]. / 'Mikrobielle Luftverunreinigungen'. Emissionsquellen und -minderungsmassnahmen.

Already in the planning or monitoring phase, measures have to be realized for the reduction in emissions of bio-aerosols in biological waste treatment plants. For this purpose, operation-related measures (in open and partially closed plants) and technology-related measures (in closed plants with automatic ventilation) can be taken. A survey on the currently feasible measures for reduction is submitted. The realization has to be adapted to the individual site and type of plant. An enormous demand for examinations is envisaged in order to enable a qualified assessment of the emission behaviour of diverse waste air systems or process steps.

[Detection of airborne cultivatable microorganisms from compost sites--emissions and imissions]. / Erfassung von luftgetragenen kultivierbaren Mikroorganismen aus Kompostierungsanlagen--Emission und Immission.

The statement presented here gives an overview and assessment of the procedures and concepts currently used for the collection and determination of airborne, culturable microorganisms at sources of emission within composting plants and in their near vicinity. The paper focuses on "classical" methods, which involve cultivation as an intermediate step for the determination of viable, airborne microorganisms. The theoretical and practical requirements on such methods are discussed. Results and experiences from recent investigations are described.