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Ochratoxin A (OTA) is a mycotoxin produced by several moulds of Aspergillus and Penicillium genera on many agricultural commodities used for feed production. It is primarily a nephrotoxic substance, but it can also cause immunosuppression in pigs and increase their susceptibility to infections. Porcine mycotoxic nephropathy (PMN) is a chronic poisoning associated with OTA exposure. The aim of this paper was to evaluate the content of OTA in pig kidneys which were examined during the regular monitoring of the safety and quality of food of animal origin in the Czech Republic during the years 2012–2021. Results revealed no differences between the monitored years or age categories and no differences between the year seasons, which could have an influence on the storage conditions and thus feed contamination by moulds. The values found in this study are low and similar to those assessed in several studies performed in other European countries and do not exceed the limits set in their legislation, which means these organs do not pose a risk for consumers and do not indicate PMN in monitored pigs. Czech Republic as well as EU legislation do not regulate OTA levels in the food of animal origin yet.

The wide use of cell technologies in clinical practice requires a large amount of cell material, which has led to improvement in culture conditions, making it possible to obtain more cell material in a shorter period of time. Thus, the purpose of our paper was to study the effects of different concentrations of an insulin-like growth factor (IGF-1), a fibroblast growth factor (FGF-2),| a growth hormone (rhGH), and Biolaminin 521 LN (LN 521) on the proliferative activity and genetic stability of stem cell cultures derived from the cat bone marrow, adipose tissue, and myocardium. Cell cultures for the experiment were obtained from the adipose tissue, bone marrow, and myocardium of a cat. Differences were found in the effects of the various growth promoters on the proliferative activity of cells in the culture. The IGF-1 demonstrated a positive effect on the proliferative activity of all cultures. The addition of the rhGH to the bone marrow-derived cell culture increased the size of the cells and decreased the proliferation index relative to the control group. The addition of the growth factors to the culture medium did not significantly increase the number of cells with altered karyotype in any of the cultures relative to the control group.

Many important discoveries have been made in the field of nanotechnology in the last 40 years. Since then, nanoparticles became nearly ubiquitous. With their spreading use, safety concerns have warranted extensive research of nanotoxicity. This paper offers information about the occurrence, transport, and behaviour of metallic nanoparticles in the aquatic environment. It further summarizes details about parameters that dictate the toxicity of nanoparticles and discusses the general/common mechanisms of their toxicity. This review also focuses on fish exposure to nanoparticles, including the possibility of trophic transport through the food chain. Information on some of the most frequently used metallic nanoparticles, such as silver, gold, and titanium dioxide, is further elaborated on.

Angiomyolipoma is an extremely rare neoplasm in animals. It belongs to a group of perivascular epithelioid cell tumours (so-called PEComas). This study reports a case of primary soft tissue angiomyolipoma in a dog with some unusual clinical features. A 4-year old female Labrador dog with a rapidly growing pelvic tumour measuring 30 × 20 cm with a short history of 12 weeks was presented. The tumour was well-circumscribed and pushed into the right vaginal wall and into the perineum. The tumour was completely surgically excised. An extensive histological examination was performed, including immunohistochemical analysis. Histology revealed a mesenchymal neoplasm consisting of three tissue components – mature adipose tissue (which dominated), vessels, and smooth muscle (spindle) cells. The lesion showed positive immunohistochemical staining with smooth muscle actin and desmin in mature-appearing smooth muscle cells and S-100 protein positivity in adipocytes. The diagnosis of angiomyolipoma must be considered in different types of benign and malignant tumours of various lineages. The diagnostic approach to soft tissue tumours, including angiomyolipoma, requires optimal processing and sectioning of resected specimens. This paper is believed to be the first case of primary soft tissue angiomyolipoma reported in the veterinary literature.

Helicobacter pylori (H. pylori) is found worldwide, and it is estimated that more than 50% of the population is infected. The presence of H. pylori in the gastrointestinal tract is associated with the occurrence of serious human diseases such as chronic gastritis, duodenal ulcer or gastric cancer. Therefore, the International Agency for Research on Cancer classified H. pylori as a Group 1 carcinogen in 1994. The routes of H. pylori transmission are not yet precisely known. Many authors agree that humans may become infected by H. pylori through foodstuffs. Milk is one of the basic foods of people of all age categories. Helicobacter pylori has been isolated from milk derived from various livestock, such as cows, sheep, goat, camel or buffalo milk. This review research paper indicates that H. pylori may be present in different types of foods and suggests that consumption of milk and selected dairy products may be a source of H. pylori infection for humans.

Severe spinal cord injury (SCI) causing significant morbidity and mortality remains one of the most challenging problems in both human and veterinary medicine. Due to the restricted regeneration potential of the central nervous system (CNS) in mammals, the neurological deficit caused by spinal cord (SC) injury is permanent, and no therapeutic measures are able to completely restore neurological functions either in primates or in non-primate animals with traumatic tetraparesis/tetraplegia or paraparesis/paraplegia. The constant progress in the understanding of pathophysiologic events developing after spinal cord trauma constitute an unremitting inspiration for neuroscientists and health care professionals to test novel medicaments and treatment strategies to cope with this situation. Recent experimental studies and preclinical trials have delivered promising results. The aim of this review is a presentation of generally accepted methods of management of dogs with SCI as well as a report on new therapeutic modalities, and comment on their potential for clinical translation. The research strategy involved a search of PubMed, Medline, and ISI Web of Science from January 2010 to December 2018 using the terms “spinal cord injuryˮ and “management of spinal traumaˮ in the English language literature. References from selected papers were also scanned and evaluated for relevance.

The aim of the study was to identifyCampylobacterspecies in a group of patients with chronic gastrointestinal problems and to investigate the relationship between the presence ofCampylobacterspp. in stool samples and as well as the severity of chronic enteropathy. Twenty-six dogs with chronic gastrointestinal problems were included in the prospective study. Each research subject had their stomach, duodenum, ileum, and colon examined endoscopically. A histopathological examination of the obtained biopsy samples was then performed, excluding other potential diseases. Stool samples were collected and then examined for the presence ofCampylobacterspp. To evaluate the relationship betweenCampylobacterspp. occurrence and the intensity of chronic enteropathy, patients were divided into two groups; animals in the first group presented with no to mild inflammation whereas research subjects in the second group suffered from moderate to severe inflammation. Subsequently, the patients were divided based on positive or negative test results forCampylobacterspp. cultures. No significant relationship between the presence ofCampylobacterspp. in stool samples and chronic enteropathy was found. In contrast to other previously published papers, our study showed a lower occurrence ofCampylobacter upsaliensis.

Tissue engineering promotes tissue regeneration through biomaterials that have excellent properties and have the potential to replace tissues. Many studies show that bacterial cellulose (BC) might ensure tissue regeneration and substitution, being used for the bioengineering of hard, cartilaginous and soft tissues. Bacterial cellulose is extensively used as wound dressing material and results show that BC is a promising tissue scaffold (bone, cardiovascular, urinary tissue). It can be combined with polymeric and non-polymeric compounds to acquire antimicrobial, cell-adhesion and proliferation properties. To ensure proper tissue regeneration, the material has to be: biocompatible, with minimum tissue reaction and biodegradability; bio-absorbable, to promote tissue development, cellular interaction and grow; resistant to support the weight of the newly formed tissue. Its versatile structure, physical and biochemical properties can be adjusted by adapting the bacteria culturing conditions. The main biomedical applications seem to be as hard (bone, dental), fibrocartilaginous (meniscal) and soft tissue (skin, cardiovascular, urinary) substituents. This paper reviews the current state of knowledge, challenges and future applications of BC and its biomedical potential in veterinary medicine. It was focused on the main uses in regeneration and scaffold tissue replacement and, although BC showed promising results, there is a lack of successful results of BC use in clinical practice. Most studies were performed only at experimental level and further research is needed for BC to enter clinical veterinary practice.

A lesion of sacrococcygeal spinal nerve roots forming a structure that resembles a horse's tail results in the development of clinical entity identified as the cauda equina syndrome (CES). The disease can evolve slowly and symptomatology can be incomplete, but the fully developed CES is characterized by pain and altered sensation in the pelvic extremities, tail, perianogenital region, paresis or plegia of hind limbs, incontinence and impotence. Major causes of CES in dogs are degenerative changes of the lumbosacral vertebral column, haematoma, inflammation, neoplasm or trauma. The diagnosis is based on history, clinical presentation, neurological symptomatology, spinal röntgenography, computed tomography, and magnetic resonance imaging. In animals experiencing initial episodes of CES, conservative therapy can be attempted. But the only rational treatment of patients with severe neurological deficit is surgical decompression of the neural structures. The outcome depends on the underlying aetiology and the degree of sensory, motor and autonomic dysfunction. Canine and porcine experimental models mimicking the CES showed the involvement of intrinsic spinal cord structures. This points out the need for an early diagnosis followed by aggressive management before irreversible neuronal lesions develop. The search strategy involved the PubMed, Medline, Embase and ISI Web of Science from January 2000 to August 2017 using the terms 'cauda equina syndrome' and 'lumbosacral stenosis' in the English language literature; also references from selected papers were scanned and relevant articles included.

Parenteral iron administration has been a common practice for the prevention of iron deficiency in newborn piglets. The efficacy and safety of this method require reexamination due to the introduction of new genetic lines and management changes in swine production. The aim of this article was to review current knowledge on this method of anaemia prevention in piglets. Iron requirements, iron sources, and the mode of action, dosage, and negative effects of iron dextran injection are discussed. The paper also reviews methods for evaluating the efficacy of iron administration in piglets.

The time around calving in highly productive dairy cows is a critical period in terms of their metabolism, which is connected with high demands of the foetus as well as with the onset of lactation. Retained placenta in cows may have multifactorial aetiology, but in herds which are free from infectious diseases, the most important reasons are; periparturient metabolic changes and disturbances to the internal balance and stress. During the periparturient period, the most important factor causing immune suppression and hypotony of uterus in cows is metabolic stress due to hormonal and nutritional factors, including metabolic fluctuations, negative energy balance, as well as shortage of proteins, minerals, vitamins and antioxidants. This metabolic stress as a result of an imbalance in the internal metabolic homeostasis activates the hypothalamic-pituitary-adrenocortical axis (HPA) and increases serum corticosterid (cortisol) concentration, especially on the day of calving. Cortisol is a powerful immune suppressive factor that causes depression of leukocyte proliferation and their functions. The periparturient metabolic stress may also stimulate the production of catecholamines, especially adrenalin. Elevated levels of adrenalin activate adrenoreceptors of the myometrium, which in turn cause hypotony or atony of the uterus at calving in cows. Elevated levels of cortisol and adrenalin may significantly inhibit the rejection and expulsion of foetal membranes in cows, resulting in an increased incidence of their retention. These important mechanisms for placental retention in highly productive dairy cows often have primary nutritional metabolic aetiology, but they also occur during secondary metabolic disturbances and metabolic stress during calving. This metabolic and immunological aetiology and pathogenesis of retained placenta usually occur in highly productive periparturient cows on dairy farms in the absence of bovine infectious diseases, which can couse placentitis, stillbirth and the infectious origin of foetal membrane retention. The paper presents the most important metabolic, mineral and immunological disturbances as conditions for retained placenta in dairy cows. It also shows different methods of herd monitoring, important examples of clinical and laboratory diagnostics, and methods of nutritional and veterinary prevention of this health problem in dairy cows.

Vacha, J. V. Znojil, Jirina Hola, J. Dungel: Iron Kinetics in Individual Bone Marrow Regions (Bones) in Mice. Acta vet. Brno, 53, 1984: 119-136. The course of concentration of labelled iron (.oFe) after its intravenous admi­ nistration was observed experimentally in plasma, erythrocytes, spleen and indi­ vidual bones (here in haem and non-haem fraction separately). The optimum number of iron compartments in individual bones and the constants characterizing the flow through these compartments was then sought by means of mathematical mo­ delling. It was possible to type the compartments by comparing flows into the bones of various types and to try to find their physiological interpretation. The following compartments were determined (apart from the circulating plasma and erythrocytes): 1. An iron pool with a half-time of efflux of several tens of minutes to several hours, probably corresponding to the iron contained in the interstitial marrow and splenic spaces, in a rapidly assumed equilibrium with plasma; 2. An iron pool with a half-time of efflux about nine hours, corresponding to the iron incorporating into haem or deposited in the pre-haem pool in erythroblasts; 3. An iron pool with a half-time of efflux of about 1-3 days, having the character of stores, probably deposited both in erythroblasts (half-time 3.15 days) and in cellular elements of the haematopoietic microenvironment (half-time 1.65 days); 4. An iron pool bound in the long-term in the teeth. The bones could be divided according to the represen­ tation of compartments into three groups: 1. Long bones and spine, in which inten­ sive erythropoiesis takes place, exhausting the interstitial pool and consuming most of the storage iron pool; 2. Acral bones and the skull, with minimal erythropoiesis, in which the dynamics of the interstitial and storage iron pool localized beyond erythroblasts predominantly occurs; 3. Thoracic bones, representing a group inter­ mediate between the two former ones in several aspects. - The model is also in accordance with the independently (chemically) ascertained iron content in the bones. The results represent a considerable refinement of our general model of ferro­ kinetics in mice (Vacha et al. 1982) and provide a firmer basis for the study of ferrokinetics in bone marrow under pathological states of erythropoiesis. Studies of this type are an inevitable prerequisite of correct interpretation of the labelled iron incorporation method as an indicator of the intensity of erythropoiesis. Ferrokinetics, iron compartments in bones of various types, haem and non-haem iron, .oFe-incorporation method. Knowledge of the internal exchange of iron in an organism is a prerequisite for the correct interpretation of the commonly used method of labelled iron incorporation into the erythropoietic organs. The internal iron kinetics in mice are, however, known only incompletely as yet. In our previous paper (Vacha et al. 1982) we proposed a model of the ferrokinetics of mice encompassing the sizes of basic iron compartments in the erythropoietic organs, the blood and the liver and the extent of flows between them. The employment of a model of ferrokinetics for the study of erythro­ poiesis presupposes, however, an analysis of iron compartments in the bone marrow more sophis­ ticated than that which can be made in such a general model. The basic idea of the present paper

The objective of this work was to study the changes in chlorophyll contents in green teas stored under different conditions and to suggest the most suitable way and length of green tea storage. A secondary aim was to evaluate the possibility of using the chlorophyll content as an indicator of tea freshness. Samples for analysis were obtained from a range of tea-growing regions. Chlorophyll contents were measured using UV-vis spectrophotometry. Chlorophyll absorbance was measured at 642.5 and 660 nm. Prior to storage, chlorophyll concentration in teas ranged from 1.12 to 1.89 mg per 1 g of tea, which corresponded to data given by other authors. Storage was found to lead to a decline in chlorophyll concentration. From the 6th month of storage onwards, a significant decrease (P < 0.05) was observed in all samples, irrespective of storage conditions. Based on the findings of this study, original and metal packaging can be considered the most chlorophyll-friendly. On the other hand, glass and paper packaging stored in direct daylight were found to have the strongest impact on chlorophyll concentration. Overall, it can be concluded that the storage time can have a strong influence on oolong green tea colour as an important qualitative variable. Thus, the shelf-life of tea should be shorter than the 12 months claimed by most manufacturers. Once the relationship between changes in chlorophyll concentration and length of the storage period had been established, the chlorophyll content was suggested as an indicator of the storage time (freshness) of a tea substance.

The colour and viscosity of egg yolk are among major indicators assessed by consumers and food technology. This study attempts to evaluate the colour and viscosity of yolk in laying hens’ eggs after the addition of dried beetroot (Beta vulgaris L. ssp. esculenta var. rubra) at the amount of 1% and 2% per feeding dose (in July and August 2012). The experiment was performed on 24 hens that were divided into three groups of 8 laying hens. The preparatory phase lasted one week (standard diet), followed by four weeks during which experimental layers received a diet enriched with beetroot. Then, all layers were fed a mixture without beetroot for the following four weeks. Eggs were collected during the whole period of 8 weeks. In total, 30 eggs from each group were subjected to analysis. The colour of eggs was determined using spectrophotometry, by the Colour-guide sphere spex portable colorimeter. The results showed a significant (P < 0.05) increase in value L* of yolk colour in experimental groups whereas values a* and b* (indicators of the international colorimetric scale CIELAB) did not vary significantly. Similarly, specific purity C*ab did not show a significant difference (P < 0.05) between the control and experimental groups. The egg yolk viscosity was lower in experimental groups compared to the control group but the difference was not significant. The addition of dried beetroot at the amount of 1 and 2% per feeding dose had no effect on colour and viscosity. This paper supported the null hypothesis that the addition of dried beetroot to the feeding dose at the amount of 1% and 2% has no effect on the colour and viscosity of egg yolk. Antioxidants, CIELAB, Beta vulgaris, poultry nutrition, food technology The colour and viscosity of egg yolk are among major indicators assessed by consumers when they evaluate the quality of eggs. Egg yolk contains 35% of lipids, 16% of proteins and most 0.2% of lipophilic pigments including carotenoids (Kriz 1997). All egg yolk components contribute to its final colour. The effect of diet administered to laying hens is manifested by a number of colour tones, ranging from light yellow to dark orange (Lomakina and Mikova 2006). The colour of egg yolk is also affected by carotenoids, particularly lutein and zeaxanthin, and xanthophyls with their antioxidant effects. Beta vulgaris L. ssp. esculenta var. rubra contains among others also flavonoids, betanin, quercetin, and polyphenols (Lichtenthaler et al. 2005). The term „chromaticity“ has been used to describe the properties of the source of light and express colour properties (spectral composition). Unlike the subjective perception of colour, colorities can be compared using spectrophotometric instruments with defined parameters (Dvorak et al. 2009). The results of reflection and absorption are expressed using the international colorimetric scale CIELAB (Commission Internationale de l’Eclairage 1986) which is based on differences between three elementary complementary colour pairs: red-green (value a*), yellow-blue (value b*) and black-white (value L*). The third indicator, specific lightness L*, is the function of reflectance, i.e. the ratio between the intensity of reflected light and the intensity of incident light from 0 (black) to 100 (white). The other indicators are defined by ACTA VET. BRNO 2014, 83: 039–044; doi:10.2754/avb201483010039 Address for correspondence: Vladimir Kopřiva Department of Biochemistry and Biophysics Faculty of Veterinary Hygiene and Ecology University of Veterinary and Pharmaceutical Sciences Brno Palackeho tř. 1/3, 612 42 Brno, Czech Republic Phone: +420 541 562 610 E-mail: koprivav@vfu.cz http://actavet.vfu.cz/ calculations: The distance between the two points represented by ΔE* (CIE total colour difference); specific purity C*ab (the chroma of the colour) is the value by which particular colority differs from grey; the specific angle of the colour tone hab (hue) is described as the name of colority in relation to colour (Dvorak et al. 2009; 2012). The diet composition is a major factor affecting the colour of egg yolk. Viscosity as a physical indicator depends on the age of an egg, storage temperature, the pH value, specific weight, water content and egg stress. When assessing the viscosity of the whole egg, one should particularly consider the viscosity of egg yolk since it is usually × 8 greater than the viscosity of egg white (Simeonovova et al. 2003). The aim of this study was to find if the addition of beetroot to poultry feed affects yolk colour detected by the SIELAB system, and whether the addition of beetroot to the diet of laying hens affects the viscosity of egg yolk. Materials and Methods Experimental design The experiment was performed in the accredited enclosure of the Faculty of Veterinary Hygiene and Ecology, University of Veterinary and Pharmaceutical Sciences Brno. The laying hens were reared using enriched cage technology. The experiment was performed (in July and August 2012) on 24 Isabrown hybrid hens aged 28 weeks that were divided into three groups of 8 laying hens. The groups of hens were housed in separate cages. During the experiment, laying hens received a complete commercial feeding mixture ad libitum (crude protein 159.0 g·kg-1, fat 25.8 g·kg-1, fibre 26 g·kg-1, ash 104.5 g·kg-1, Ca 33.2 g·kg-1, P 4.9 g·kg-1, Mg 3.1 g·kg-1 and metabolic energy 11.7 MJ·kg-1). Water was administered continually ad libitum using automatic feeders. Two experimental groups received homogenized dried beetroot (Beta vulgaris L. ssp. esculenta var. rubra) at the amount of 1% (group 1) and 2% (group 2) in their feeding mixture. The last group was control that received only the feeding mixture without the addition of beetroot. One week before the start of the experiment the hens were fed a standard diet. This week was followed by 4 weeks of diet enriched with beetroot. Afterwards all layers were fed a mixture without beetroot for the following 4 weeks. Eggs were collected during the whole period of 8 weeks. In total, 30 eggs (that were allowed to stand at 6 °C for 96 h) from each group were subjected to analysis. Methods Egg yolk colour was determined first, followed by determination of yolk viscosity. Prior to measurement, egg yolk was separated from egg white and placed on a Petri dish with a diameter of 50 mm. Then, it was mixed and covered using a thin plastic foil for wrapping food. The colour was determined using a spectrophotometric method in the CIELAB system, using the Colour-guide sphere spex portable spectrophotometer (Byk Gardner), without gloss, with the following set-up: spherical geometry d/8°, the source of light D65, standard observer’s angle 10°, and the diameter of aperture 8 mm. All measurements were carried out × 3; the mean of all three measured values was calculated to give the resultant value. In order to compare the results, the mean values of L*, a*, b* coordinates were used to calculate further parameters of the CIELAB system. The distance between the two points ΔE* (CIE total color difference) was calculated according to the following formula: DE* = [(DL*)2 + (Da*)2 + (Db*)2]1/2. Chroma C*ab is the value by which the particular colority differs from grey; it was calculated according to the following formula: C*ab = (a* 2 + b*2)1/2. Hue hab is described as the name of a colour. Its value is calculated according to the following formula: hab = tg -1 (b*/a*). Egg yolk viscosity was determined using the Rheo-viscometer. Samples in a cuvette KZ 100 were allowed to reach 20 °C. The determination of viscosity of substances in a range of 0.05 to 106 Pa·s. Due to the size of the cuvette, two homogenized and tempered yolks were measured at the same time. The sample was poured into the cuvette and mixed × 10 by moving a ball inside the cuvette. A total of 15 samples from each group were analysed on a particular date of collection. Viscosity was measured × 3 for each sample of yolks and the arithmetic mean obtained from three measurements was used in further evaluation. A period of 40 s was allowed between individual measurements. Statistical analysis Data were statistically processed for each indicator (L*, a*, b*) and viscosity (Table 1 and Figs 1–3). The dependence of egg yolk colour indicators and viscosity on the addition of beetroot in a feeding dose was tested by using one-factor ANOVA (MS EXCEL 2003). Dependences of the above-mentioned factors on the laying period were based on the mean values in individual collection cycles using two-factor ANOVA. Since we did not expect large differences among experimental groups, null hypothesis (feeding the laying hens with a diet supplemented with beetroot does not affect colour or viscosity of egg yolk) was established for the level of significance α = 0.05. 40

This paper describes the morphologic structure of chickens’ livers as a result of restrictive feeding in variable intensity. This experiment was performed on 96 chickens of two hybrids (Ross 308 and Cobb 500) of both sexes. The chickens were divided into 3 groups (control group and two groups with restricted feeding). There were 8 chickens in each group. After hatching, the control group was fed a standard mixture ad libitum. The first restricted group was fed 80% ad libitum from the age of 2 weeks. The second restricted group, from the age of 2 weeks, was fed 65% ad libitum. The experiment lasted for 5 weeks. The liver samples were obtained by necropsy, then weighed, and after this processed by standard histological methods. The slices of liver were stained with haematoxyline-eosin. The liver samples were evaluated both objectively and subjectively. In the control group, steatosis and marginal pathological acute venosis hyperaemia were found. In the group with 80% restriction, perilobular steatosis was observed. In the group with 65% restriction, centrolobular steatosis was observed. Controlled measurement of the average hepatocyte thickness in the individual sample groups was performed using an objective method. The hepatocyte diameter thickness of males was higher than the hepatocyte diameter thickness of females, which was significant (P < 0.001). This is the first work providing the possibility of morphometrical verification of the time factor including other morphological criteria.

The present paper extends the current knowledge of biochemical and mineral profiles in selected species of feathered game such as helmeted guineafowl (Numida meleagris), common pheasant (Phasianus colchicus), Japanese quail (Coturnix coturnix japonica), chukar (Alectoris chucar), grey partridge (Perdix perdix) and wild turkey (Meleagris gallopavo) reared in the Czech Republic and throughout Europe. The work evaluates potential interspecies differences among the values and compares the findings with the results reported for related domesticated species of birds. Indicators investigated in the individual species of feathered game such as total protein (TP), glucose (GLU), total cholesterol (CHOL), triacylglycerides (TAG), uric acid (UA), alanine aminotransferase (ALT) and Ca, P, Mg, Na, K, Cl varied in ranges typical for layers and broiler chickens. Except for wild turkey where mineral metabolism values reached or even exceeded the upper limit of the physiological range, which may indicate a specific interspecies difference. However, with regard to the wide physiological range of biochemical indicators in the blood of birds, significant (P ⪬ 0.05) and highly significant (P ⪬ 0.01) differences in most mean values for the monitored indicators can not be evaluated as interspecies differences.

The present paper extends the knowledge of selected blood indicators in six species of feathered game such as helmeted guineafowl (Numida meleagris), common pheasant (Phasianus colchicus), Japanese quail (Coturnix coturnix japonica), chukar (Alectoris chucar), grey partridge (Perdix perdix) and wild turkey (Meleagris gallopavo), reared in the Czech Republic and Europe. The red blood cell (RBC) values in individual species of feathered game ranged as follows: the mean RBC counts varied in a range of 2.10 - 2.58 T/l; haematocrit values were in a range of 0.33 - 0.39 1/1; haemoglobin concentration varied in a range of 101.20 - 129.13 g/l; mean corpuscular haemoglobin concentration (MCHC) ranged from 28.63 to 35.44%; mean corpuscular haemoglobin (MCH) varied in a range of 46.15 - 62.07 pg and the mean corpuscular volume (MCV) was 147.66 - 178.20 fl. Except for the total concentration of haemoglobin and the mean corpuscular haemoglobin concentration, no significant differences were found between the mean values of RBC values in individual species of feathered game. However, some differences between species were found in white blood cell (WBC) values, ranging as follows: the total WBC count ranged from 13.55 to 26.90 G/l, basophil granulocytes from 0.011 to 0.110 G/l, eosinophil granulocytes from 0.011 to 0.593 G/l, neutrophil granulocytes from 1.868 to 6.661 G/l, lymphocytes from 10.937 to 20.922 G/l and monocytes from 0.034 to 0.199 G/l. Most values showed significant (P ⪬ 0.05) to highly significant (P ⪬ 0.01) interspecies differences. The main goal of the study was to investigate selected haematological indicators in six major species of feathered game.

The seeds of sweet lupine cultivars (Lupineus species) have been used with increasing frequency as a source of proteins replacing proteins of animal origin or soybean in feed compounds. The seeds of sweet lupine cultivars contain an average of about 33-40% crude protein in dry matter, the lipid content ranges from 5 to 13%. The profile of amino acids is relatively beneficial, however, with slightly lower content of lysine (1.46%) and methionine (0.22%). The main storage carbohydrates in the seeds are the β-galactans that comprise most of the cell-wall material of the kernel and the cellulose and hemicellulose of the thick seed coats. Lupine seeds contain about 40% non-starch polysaccharides (NSP) and a negligible amount of starch. High coefficient of protein digestibility (> 90%) has been reported in sweet lupine cultivars, but lower digestibility of energy (~ 60%). Lupineus species contain negligible amounts of trypsin inhibitor so they do not require preheating before being used as an ingredient in feeds for monogastric animals. The efficiency of lupine seeds in the diets for pigs is characterized by controversial results of growth and feed conversion under both mechanical treatment and enzyme supplementation. The results of production efficiency obtained at the testing of lupine cultivars (L. albus, L. angustifolius) for pigs were beneficial at supplementation of the lacking nutrients or dehulling. The submitted paper summarizes national as well as foreign knowledge of the nutritive value of cultural lupine seeds, and deals with the possibility increasing nutrition and production efficiency in the diets for pigs. Lupineus sp., crude protein, oligosaccharides, nutrients, antinutritive effect, enzymes, production efficiency Lupine has been included among eight potential vegetable sources of protein for the use in feed and food that replace proteins of animal origin in the diets (Dijkstra et al. 2003). The yield of 3,000 to 5,000 kg/ha seeds results in a potential production of up to 2,000 kg/ ha protein. The world producer and exporter of lupine seed is Australia, whose production represents 80-85% of global production, of which 90-95% is exported (White et al. 2007). In 20052006 Australian production exceeded 1 million tons of low-alkaloid lupine, of which more than 300,000 tons were for their own consumption and the rest was exported to the whole world including Europe. Approximately 40% of the exported lupine seed is used as feed for dairy and meat-type cattle, 40% as feed for pigs feed and the rest is equally distributed for the nutrition of sheep, goats and poultry (White et al. 2007). In Europe, the main growing areas can be found in Germany, France, Benelux, Spain, Poland, Ukraine and Russia. At present, so called “sweet” lupine variants are being cultivated, in which the content of bitter substances does not exceed 0.05%. A very low content of nutritive-active factors, low allergenicity and a positive impact on prevention of cardiovascular diseases (hypocholesterolaemic effects and anti-hypertensive activity) have been reported in these cultivars, which leads to their increased use in feed and food industry (Martins et al. 2005). In the Czech Republic we lack precise data on lupine cultivation. According to the estimation of the company Agritec Sumperk Ltd., the lupine growing area could reach up to 7,000 ha, with an average yield of about 3,100 kg/ha on some farms. White lupine (L. albus L.), narrow-leafed lupine (L. angustifolius L.) and yellow lupine (L. luteus L.) are the most frequent species cultivated in the Czech Republic. At present, most of the lupine production is intended as components of feed compounds; whilst the market with ACTA VET. BRNO 2009, 78: 399-409; doi:10.2754/avb200978030399 Address for correspondence: Ing. Bohumila Pisařikova Veterinary Research Institute Hudcova 70, 621 00 Brno, Czech Republic Phone: +420 533 331 616 Fax: +420 541 211 229 E-mail: pisarikova@vri.cz http://www.vfu.cz/acta-vet/actavet.htm lupine seed for food industry is still being developed. The importance of lupine cultivation can also be seen in the EU policy supporting the HEALTHY-PROFOOD research project (Anonymous 2008). Species, cultivation, physical and chemical characteristics and basic analyses The genus of lupine (Lupineus L.) belongs to the family Fabaceae fabaceous (leguminous), subfamily Genisteae (Sikula and Zubrický 1964). The genus comprises 200-300 species of which only five have been used as cultural plants. In the Czech Republic, white lupine (L. albus), narrow-leafed lupine (L. angustifolius) and yellow lupine (L. luteus) are currently used for both feed and food purposes. The thin root system with its deep penetration of up to 20 cm bellow the surface, enabling nutrient uptake from deeper layers, are some of the major advantages of lupine plants (Anonymous 2007). Lupine growth is promoted by bacteria in the root system that can fix nitrogen from the atmosphere and transform it into more readily available ammonia. Lupines are nodulated by soil bacteria that can, under favourable conditions, provide all the nitrogen needed. The nodules are able to fix 160-200 kg nitrogen/ha during the vegetative period, of which about one half is left in the soil, thus fertilizing plants the following year. Lupine is also able to receive sparingly soluble phosphates from the soil. Potassium is not a limiting element, either. Nitrogen fertilizers are only applied in very low productive soils at the rate of 10-15 kg/ha. Increased sensitivity to the complex of root and cervical diseases and susceptibility of stems to mycoses has been observed during the vegetative period. The complex Botrytis cinerea, Fusarium oxysporum and Phoma medicanidis have the worst effects, followed by anthracnosis caused by the fungus Colletotrichum lupinei (Plaskowska et al. 2000; Nirenberg 2002). White lupine (L. albus) is an autogamous plant. It reaches the height of 0.75–1 m. The flowers are blue or white. The seeds are large, cream in colour with a circular flattened shape, and with a 1,000-seed weight of 350-400 g. Narrow-leafed lupine (L. angustifolius) is 0.40-1 m high; the flowers are white and blue and the plant has considerably narrow leaflets of palmate compound leaf. Yellow lupine (L. luteus) is predominantly autogamous, 0.40-0.75 m high with yellow or orange flowers. Both the latter species have round seeds with seed coat colour from white and yellow to brown, mostly with brown or black marbling. The 1,000-seed weight is 130-180 g. The hull accounts for 22% of lupine seed weight (Anonymous 2007). Proteins and amino acids The seeds of sweet lupine species (L. albus, L. angustifolius, L. luteus) contain 28 to 48% (an average of about 33-40%) crude protein in dry matter, which depends on the species and climatic conditions (Hove 1974; Green and Oram 1983; Sousa et al. 1996; Linnemann and Dijkstra 2002; Sujak et al. 2006). Great differences in the protein content (from 317.1 g/kg cv Boruta to 458.8 g/kg cv Juno in dry matter) were found in the most frequently cultivated lupine species in Europe by Strakova et al. (2006). Seed analysis of L. luteus cv Juno showed crude protein contents in the range of 39.8 to 48.2% in dry matter (Roth-Maier and Kirchgessner 1993). Variability in protein content is determined by genetic and environmental effects and the year of harvest (Zdunczyk et al. 1996) (Table 1). The profile of amino acids is characterised by a lower level of methionine (~ 0.2% of the seed, 0.66 g/16 g N), cysteine (~ 0.4% of the seed, 1.36 g/16 g N) and lysine (1.46% of the seed, 4.75 g/16 g N) (Petterson 2000). Threonine level reaching 1.15% is low, and tryptophane content with 2.76 g/kg is also low compared to soy (Simon and Jeroch 1999). In contrast, arginine content is markedly higher, reaching 31.0 to 48.0 g/kg (Suchý et al. 2005). The highest level of amino acids within the crude protein 400

Pavlik A., R. Filipcik, P. Jelinek, M. Bjelka, Z. Havlicek, J. Subrt: Indicators of the Internal Environment of Beef Cattle during Fattening and their Correlation to the Quality of Beef. Acta Vet. Brno 2008, 77: 539-546. The objective of the study was to determine selected indicators of the internal environment in individual cattle categories during the period of fattening to evaluate the relation of the individual blood components to the growth intensity and qualitative indicators of the meat. The trial involved half-siblings - bulls, heifers and steers housed in one stable. The threesome always had a common father. During the trials the live weight was monitored and blood samples were taken on days 310, 403, 448, 479, 512, 554 and 618 of age. The concentrations of total proteins, glucose, total cholesterol, urea and catalytic concentrations of ALP, ALT, AST, calcium, inorganic phosphorus, magnesium and copper in the blood plasma were assessed. In all cases the levels of indicators of the internal environment were within the range of the physiological values for the respective age category. Compared to bulls and steers, the concentration of total plasma cholesterol of the heifers was significantly higher (p < 0.05 and p < 0.01, respectively). The last samples were taken prior to slaughter of the animals and the individual components were correlated with the nutritional values and meat indicators; the correlation between the amount of glucose and urea in the blood serum and the tenderness of the meat was significant (p < 0.05 and p < 0.01). The correlation between the concentration of proteins in the blood plasma, the level of nitrogen or total protein in the muscles, the water-holding capacity and meat tenderness was positive. The study provides a basis for implementing helpful adjustments in current cattle management practises. Blood indicators presented in this paper can be added to existing knowledge to benefit veterinary research and clinical evaluations. Growth, cattle, blood, enzymes, total proteins, glucose, phosphorus, magnesium, copper, cholesterol, water-binding capacity, shearing tests

Strakova E., P. Suchý, M. Sugerkova, M. Machacek: Positive Effect of Plant-Based Diet on the Performance and Health of Laying Hens. acta v et. Brno 2007, 76: S31-S37. an entirely plant-based feed mixture was prepared to minimize the potential risk of transmitting prion infections through animal feed. It consisted of two protein components (soya extracted meal and lupin seed meal) and was compared with a diet containing animal protein components (fish meal). the objective of the present work was to prepare and verify the effect of the entirely plant-based diet on the performance, selected qualitative indicators of eggs and the hens’ state of health. a specially prepared feed mixture containing fish meal was used as a control diet. the tested plant-based diet administered to experimental hens produced no negative effect on egg production variables such as number of eggs per hen or laying performance. the weight of eggs laid by experimental hens was significantly higher (61.66 g, P ≤ 0.01), as compared with the control group (60.03 g). the higher average weight of eggs in the experimental group increased the production of egg mass per layer by 540 g in 252-day experimental period. the beneficial effect of the plant-based diet was demonstrated particularly in the improved egg quality, manifesting itself in a highly significant improvement in yolk colour ( P ≤ 0.01) and a significant decrease (P ≤ 0.05) in cholesterol level in egg yolk. Haematological examination did not indicate statistical differences between experimental and control layers in red blood cell indices such as rBC, haemoglobin, hematocrit, MCHC, MCH, and MCv , while white blood cell indices showed a highly significant increase ( P ≤ 0.01) in Le in experimental hens. Biochemical tests performed in blood plasma revealed significantly lower molar concentrations of glucose ( P ≤ 0.05) and highly significantly lower levels of total lipids, cholesterol, lipoproteins (HDL and LDL) in experimental hens (P ≤ 0.01), as compared with the control group. the results of the study confirm that animal protein in diet for laying hens can be replaced by combination of vegetable protein feeds (soybean and lupin protein) without a negative impact on their performance and health, thereby contributing to solving issues of food safety (a prevention of prion infections). Hens, nutrition, egg production, egg quality, haematological and biochemical examination of blood In terms of the safety and quality of raw materials and foodstuffs originating from animals, constant effort in the area of animal nutrition goes into preventing the contamination of feeds with substances that may put human health at risk. the replacement of animal feed with plant-based feed may offer a potentially useful solution to this problem. Due to bovine spongiform encephalopathy in ruminants, a new strategy in the nutrition of animals reared for the production of feed was elaborated. First papers pointing out the option of preparing entirely plant-based diets were published by Suchý et al. (2002) although meat-and-bone meals were an integral part of feed mixtures for poultry (particularly laying hens) at that time. The effect of the diets administered to hens used for production of consumer eggs should be assessed in respect to both egg production performance and egg quality. In this respect, the composition of the egg (i.e. egg shell, egg white and egg yolk) is considered to be the basic indicator to be evaluated, together with the level of egg cholesterol, which reflects consumer health aspects, and egg yolk colour, which characterises egg attractiveness to consumers. the effect of the diets with and without animal protein (meat-and-bone meal)

Food safety and quality and animal production health have recently become major topics of veterinary research and the focus of several national, European and international grant agencies. Since 2005, the University of Veterinary and Pharmaceutical Sciences in Brno has been conducting two major projects in these areas. Both projects contribute to enhancing the quality of research at the university, and some results have been published in peer-reviewed international scientific journals included in the Citation Index database. Papers have also been published in Acta Veterinaria Brno, and those submitted for publication in 2007 form the basis for a special supplement dealing with veterinary aspects of food safety and quality, and animal production health.

Dvořak P., J. kunova, M. vodňanský: Change of Colour and pH-value in Pheasant Meat after Exposure to Ionizing Radiation. Acta vet. Brno 2007, 76: S67-S71. The purpose of this paper was to study the impact of ionizing radiation on the changes of colour and the pH-value of pectoral and femoral musculature in pheasants (Phasianus colchicus). The parameters of colour L*, a* and b* were observed before irradiation (24 hours post mortem) and after irradiation (60Co source, doses of 2.5 and 5 kGy). Reduction of the L* parameter, i.e. darkening, for all exposed samples was not significant. on the other hand, a significantly higher b* parameters for both exposed groups shown rather remarkably yellow colouring of pectoral musculature. Due to irradiation with a dose of 5 kGy a significant reduction (α = 0.05) of the average a* parameter from 8.99 to 7.10 was observed. However, the above-mentioned changes would not have any negative impact on the consumer. Irradiation, Phasianus colchicus, gamma-rays, food safety, irradiation pasteurization Food safety has significant priorities in the Eu countries. For some foods, it is difficult to guarantee the safety for the consumer. Game meat belongs to such foods. At present, pheasant meat can be encountered in retail shop network. Pheasants are mainly killed during the hunting season and hence the food hygiene measures as applied with livestock are impractical and have no significance in such cases (Winkelmayer et al. 2004). Due to this fact and due to possible gut damage by cold shots there is an increased risk of bacterial meat contamination. The use of ionizing radiation is one suitable technology that can guarantee bacterial reduction. The next task during meat irradiation is to achieve the same effect as with freezing and/or high-temperature treatment and/or chemical treatment – this means killing the microorganisms that cause meat “spoiling”. This may extend the shelf life during less demanding storage conditions. It is known that the doses of 1.5 to 2.5 kGy can extend the shelf live of cooled poultry by the factor of 2 or 3 compared to nontreated samples (kiss and Farkas 1972). Food irradiation is most popular in the uSA where this type of treatment is also used for technological purposes, for example, the colour improvement of meat products (Byun et al. 1999). In most countries, only limited kinds of foods can be irradiated, as stipulated by legislation (Directive 1999/3/EC). on one hand, meat irradiation is a promising that can prolong the shelf life, safety and wholesomeness, but on the other hand this kind of treatment results in physical-chemical changes (Dogbevi et al. 1999). Such changes may affect the nutritive value and sensory properties of irradiated meat Giroux and Lacroix 1998). The doses suitable for exposure kill the majority of microorganisms, with some exceptions; hence the irradiated meat is not fully sterilised. The common doses of gamma radiation (60Co) used for chilled poultry achieve 3 to 4 kGy while a dose of 7 kGy can be applied for chilled poultry with no significant sensory ACTA VET. BRNO 2007, 76: S67–S71; doi:10.2754/avb200776S8S067 Address for correspondence: Doc. MvDr. Petr Dvořak, CSc. Department of Biochemistry, Chemistry and Biophysics, university of veterinary and Pharmaceutical Sciences Brno Palackeho 1-3, 612 42 Brno, Czech Republic Phone: +420 541 562 608 E-mail: dvorakp@vfu.cz http://www.vfu.cz/acta-vet/actavet.htm changes (Farkas 1987, 1998). unlike poultry bred on agricultural plants, meat of free living and later on hunted pheasants can expose the consumers to a certain risk (e.g. the following types of bacteria: Erysipelothrix rhusiopathiae, Mycobacterium avium, Salmonella spp., and Listeria monocytogenes). Technology of pheasant meat irradiation can significantly reduce this risk depending upon the radiation dose. Moreover, presently available thermoluminiscent methods can prove that the meat has been really exposed to radiation (Atta et al. 2001). Colour belongs to the most significant sensory and quality indicators that the consumer can observe when shopping (Hood 1980). The darker colour of deer meat compared to livestock is caused by a higher fraction of blood in musculature (poor bleeding during hunting) and by a higher fraction of musculature pigments (Winkelmayer et al. 2004). The meat colour can be measured objectively by portable devices (Dvořak et al. 2001), for example, by the spectrophotometers with the spherical geometry d/8°, the light source D65, the standard angle of observation 10° and different diameters, e.g. 8 mm. The results are presented in the international colorimetric scale CIE L*a*b* (CIE 1986). The calibration is based upon black standard L* = 0 and white standard as L* = 100. Enzyme activity has a great significance for the next technological treatment of meat and its shelf life. The effect of ionizing radiation on LD, ALP, ACP, ALP, AST in tissues of cadaver is not substantial and it does not explain the changes of enzyme activity in the living organisms after gamma irradiation (Dvořak et al. 2006). The purpose of this work is to study the impact of ionizing radiation on the changes of colour and the pH-value of pectoral and femoral musculature in pheasants. Materials and Methods Thirty pheasant males were chosen for research. Immediately after killing and over the period of the experiment including exposure, the pheasants were cooled in the refrigerator at the temperature of 6 °C. The pheasant meat samples were stored in the thermic boxes during transport. Pectoral and femoral musculature of each pheasant was prepared 24 h post mortem and each sample was divided into three parts. Six groups, each composed of 30 samples, were formed (reference group RG, group exposed to a dose of 2.5 kGy, group exposed to a dose of 5 kGy in pectoral musculature and identically in femoral musculature). At the same time, the meat colour and the pHvalue in the test group was measured twice, i.e. before and after exposure. The reference group was not exposed, but colour and the pH-value were also measured twice at the same time as with the test group. Musculature was always measured on the fresh transverse slices. The measurement before irradiation was performed 24 h post mortem, then the samples were transported and irradiated. The samples were measured after 3 hours, i.e. 27 hours post mortem. The 60Co source with the dose rate of 2.7 kGy·h-1 (the BIoSTER veverska Bitýska industrial irradiation room) was applied for irradiation. The meat colour was measured by the spectrophotometric method in the CIELAB system by means of the portable Color Guide S-Spex Byk Gardner spectrophotometer with no gloss. Three places on the slice were always measured and the resulting value was averaged from those three measurements. The measured values were statistically treated by paired t-test and compared with the samples before and after exposure. The pH-value of meat was measured in the laboratory, dual pH-meter oRIon 250A+ with the connected puncture pH-electrode calibrated for 3 buffers (pH-value of 4.01, 7.00, and 9.01). The samples were always measured at the same place. The stabilisation time of the pH-value was defined by the device. Results and Discussion The results for the L* parameter, represented the specific brightness, are shown in Tables 1 and 2. Although the samples were irradiated the values were reduced in all cases (i.e. meat darkening) and the differences were not a significant. The changes in the red wavelength were observed for the a* parameter. This parameter can achieve the maximum of +120 in intensive red and the minimum in green-blue with a value of -80. The dose of 5 kGy significantly reduced the value from 8.99 to 7.10 in the femoral musculature (α =0.05), i.e. pink reduction. This tendency was similar to that of pectoral musculature and of both muscles for the lower dose of 2.5 kGy. However, the differences were not significant. S68

The paper presents the effects of temperature and feed composition on the stability of lipids in breast and thigh muscles of COBB 500 meat hybrids. The chickens were reared in accordance with the technological procedure. The diet of experimental groups contained Proenergol in combination with Energol; the diet of control group chickens contained Proenergol and soybean oil. In the control groups and in half of the experimental groups, the temperature was reduced in agreement with the technological procedure, while it was kept at the constant level of 31 °C in other groups. The feeding continued up to the age of 42 days. The level of lipid oxidation was determined using malondialdehyde assays. No differences between individual groups of chickens were found in the breast muscle. In the thigh muscle, the introduction of Energol resulted in non-significant differences between the control and chickens kept in heat stress conditions. Negative correlation between lipid oxidation and live weight was established in control chickens fed soybean oil (p < 0.05). This correlation was positive (p < 0.05) in young male chicks fed Energol and kept at constantly increased temperature. Malondialdehyde, feed supplement, temperature regimen, broiler chickens, breast and thigh muscles

Kubiaek O., L.Taras: Incidence of Pigeon Circovirus in Eurasian Collared-Dove (Streptopelia decaocto) Detected by Nested PCR. Acta Vet. Brno 2005, 74: 361-368. The paper describes the first demonstration of pigeon circovirus in Eurasian collared-dove (Streptopelia decaocto). Nested PCR was used to examine birds of the Columbiformes family. Pigeon circovirus DNA was identified in the bursa of Fabricius of an accidentally caught dove by nested PCR after the second reaction. Impaired feathers and other clinical signs observed could not, however, be attributed to circoviral infection. The sequence of the amplified PCR product - middle of the capsid protein gene - was compared against all available circovirus sequences. Both the nucleotide and the derived amino acid sequence have a very close similarity to pigeon circovirus sequences. It may therefore be concluded that the sample was demonstrated to contain pigeon circovirus or another very similar virus. We therefore assume that pigeon circovirus may infect doves, too. Whether it is a special subtype or whether doves may be infected by common pigeon circovirus strains cannot be concluded with certainty from the existing results.

·iroka Z. , J . Kri j t , T. Randak, Z. Svobodova, G. Pe‰kova, J . Fuksa, J . Haj‰ lova, J . Jarkovsk , M. Janska: Organic Pollutant Contamination of the River Elbe as Assessed by Biochemical Markers. Acta Vet Brno 2005, 74: 293-303. The aim of the study was to assess contamination of the River Elbe basin using selected biochemical markers. Biochemical markers selected were enzymes of the first stage of xenobiotic transformation, namely cytochrome P450 (CYP 450) and ethoxyresorufin-O-deethylase (EROD). The results were correlated with the most important inducers of the enzymes, i.e. polychlorinated biphenyls (PCB) concentrations in muscle tissue of fish, polyaromatic hydrocarbons (PAH) values in bottom sediments and 1-hydroxypyrene (1-OHPY) values in fish bile (terminal metabolite of PAH, or, rather, of one of them, i.e. pyrene), which were determined during the chemical monitoring of the River Elbe basin. The indicator species selected was chub (Leuciscus cephalus L.), which was captured at ten locations in the River Elbe basin. A comparison between the EROD activity and the CYP 450 content along the longitudinal profile of the Elbe showed a significant correlation at the level of significance of p < 0.05. The highest EROD activity levels in the liver were ascertained in Zelain (341 pmol·min-1·mg-1), Valy (263.2 pmol·min-1·mg-1) and Lysa nad Labem (179.17 pmol·min-1·mg-1). In Blanice (control location), EROD activity was significantly lower than in any of the other locations studied (p < 0.05). The study failed to produce an unambiguous proof of any correlations between detoxification enzyme activity (CYP 450 and EROD) in the liver and their two important inducers (PCB and PAHs). The possibility that other substances causing activation or inhibition of detoxification enzymes were in play is also discussed. Cytochrome P450, EROD, Leuciscus cephalus L., liver, PCB, PAH, 1-hydroxypyrene The River Elbe is one of the most important European rivers (total length 1103.5 km). Its extensive basin of a total of 148 268 km2 lies on the territory of two countries, i.e. the Czech Republic (51 336 km2) and Germany (96 932 km2). Intensive research of the Czech and German reaches of the Elbe started in 1991 under the Elbe I (1991 1994) project, and continued with the Elbe II (1995 1998) and Elbe III (1999 2002) projects. In those projects, large quantities of data regarding sources of pollution, chemical monitoring of hazardous substances in various components of the aquatic environment, water quality, etc. were collected and evaluated (Nesmurak 1994; BlaIkova et al. 1998; BlaIkova 2002). To enhance the relevance of results obtained by chemical monitoring, it is, however, also ACTA VET. BRNO 2005, 74: 293–303 Address for correspondence: ·iroka Zuzana Department of Public Veterinary Health and Toxicology University of Veterinary and Pharmaceutical Sciences Brno Palackeho 1-3, 612 42 Brno, Czech Republic Phone: +420 541 562 780 E-mail: sirokaz@vfu.cz http://www.vfu.cz/acta-vet/actavet.htm necessary to assess the effects of anthropogenic pollution of the aquatic environment on fish. One of possible ways of assessing such effects is the use of biochemical markers of contamination. They are measurable biochemical parameters responding usually to substances with the same mechanism of toxic effect. That means they are not, with some exceptions, specific for individual xenobiotic substances. Their advantage lies in their ability to provide comprehensive information on the effects of pollution, i.e. to reflect synergic or antagonistic effects of individual components contributing to pollution. In 2003, the Elbe IV Project was started. For reasons mentioned above, chemical monitoring in fish was complemented with assay of biochemical contamination markers. Attention focused primarily on the enzymes of the first phase of xenobiotics conversion, i.e. cytochrome P450 and ethoxyresorufin-O-deethylase (EROD). Cytochrome P450 is an important biochemical marker of surface water contamination with some industrial and agricultural pollutants (Stegeman and Lech 1991). It is now believed that the most useful is the 1A family of cytochrome P450 (Machala et al. 1997; Schlenk and Di Giul io 2002). The most potent inducers for that isoform are substances from the groups of polychlorinated biphenyls (PCB), polycyclic aromatic hydrocarbons (PAH), nitrated polycyclic aromatic hydrocarbons (NPAH) and dioxins (e.g. 2, 3, 7, 8 TCDD) (White et al. 1997; Nilsen et al. 1998; Jung et al. 2001; Schlenk and Di Giul io 2002). On the other hand, chronic exposure to these contaminants can cause a lack of CYP1A induction response (Brammell et al. 2004), and also assessment of CYP1A at the time of spawning can influence its level, because estrogens can decrease CYP1A induction (Elskus 2004). Male fish seem to be more sensitive to PAH and PCB than female fish (McArdle et al. 2004). The induction of the CYP1A family is mediated by the aryl hydrocarbon receptor (AhR) (Bil l iard 2002). Following its interaction with xenobiotic substances, it is carried to the nucleus where it is the cause of enhanced expression of genes for CYP1A and, subsequently, of increased synthesis of cytochrome proteins. The potential toxicity of pollutants depends on their affinity to the AhR. The CYP1As are also responsible for the metabolic activation of most of the known promutagens and procarcinogens, and its elevated levels are responsible for such negative effects as cocarcinogenesis, immunotoxicity and reproduction disorders (Lewis et al. 2003; Carlson et al. 2004). The model CYP1A activity is the enzyme ethoxyresorufin-O-deethylase (EROD), with its ability to convert substrates to products demonstrating fluorescence, which can then be measured. This enzyme is an important biochemical marker of contamination. The aim of the study presented here was to use the assessment of biochemical markers cytochrome P450 and EROD in the livers of the indicator fish species (Leuciscus cephalus L.) to evaluate contamination levels in various locations within the River Elbe basin. Results of chemical monitoring relevant for the above contamination markers are also outlined and correlated in the paper. They were PCB concentrations in chub muscle tissues, concentrations of 1-hydroxypyrene (1-OHPY) in chub bile samples (i.e. the final metabolite of PAHs, or rather of pyrene), and PAH concentrations in bottom sediments in the locations studied. Materials and Methods Animals and Sampling The chub (Leuciscus cephalus L.) was selected as the most suitable indicator species. The chub is a common freshwater cyprinid species that inhabits both clean and polluted rivers (Baru‰ et al. 1995). The fish were captured with the use of a diesel-electric generator in 10 locations in the River Elbe basin. The locations studied were Podoli and Zelain at the River Vltava, a tributary to the Elbe, and Verdek, Numaice, Valy, Lysa nad Labem, Obfiistvi, Duain and Hfiensko along the River Elbe. The control location was upstream of the Husinec water reservoir at the river Blanice in the Vltava basin (Fig. 1). The fish were captured in July 2003 (average water temperature 21.4 °C). In the control location, fish were captured in September 2003 (water temperature 15 °C). In each location, eight chub (both males and females) were captured (except in Lysa nad Labem where only 3 chub were captured). The chub 294

Tkaaikova a., M. R., Bhide I. Mikula: Asymmetric PCR-SSCP: a Useful Tool for Detection of OLA-DRB1(MHC class II) Gene Polymorphism in Slovak Improved Valachian Sheep. Acta Vet. Brno 2005, 74: 275-278. Detection of OLA-DBR1 (exon 2) gene polymorphism is presented in the paper. Rapid and inexpensive polymorphism detection method, namely, single stranded conformation polymorphism (SSCP) was assessed. Modification of the SSCP to asymmetric PCR-SSCP enabled a more simplified assay for the clustering the individuals into distinct groups (profiles) on the basis of band patterns, as only single stranded amplicons were detected. A total of 400 Valachian sheep were included in the study. In this cohort, 25 distinct clusters were noticed. Among 25 groups the frequency of k profile was the highest (28%), followed by profile e (21%), p (16 %), w (9%) and d (9%). The homologous SSCP as well as asymmetric-PCR-SSCP patterns were observed among the twins: This finding has shown the sensitivity of both methods to segregate the individuals on the basis of their allelic forms.

Kurtdede A., K. Ural : Haemobartonellosis in Cats in Ankara, Turkey. Acta Vet. Brno 2004, 73: 507-512. The paper describes four cases of feline haemobartonellosis (FH) in which anaemia and high fever had been the predominant presenting symptoms. In addition coughing was detected in two cats. FH was diagnosed on the basis of blood smear with Romanowski type stain. Blood samples were withdrawn before treatment (day 0) and after treatment (day 29). Mean WBC, RBC, Hb, PCV, MCV and MCHC values on day 0 were found as 16.2150·10 9 ·L -1 , 4.7425·10 12 ·L -1 , 7.8500 g·dL -1 , 26.2750%, 50.3575 fL and 32.5125 g·dl-1 respectively. Mean WBC, RBC, Hb and PCV values on day 29 were found as 9.2500·10 9 ·L -1 , 7.2925·10 12 ·L -1 , 11.4000 g·dL -1 and 33.75%, respectively. The abnormalities found on routine haematological examination were a mild normocyticnormochromic regenerative anaemia (cat 1 and cat 2), normocytic-hypochromic anaemia (cat 3) and macrocytic-normochromic anaemia (cat 4), mild eosinophilia (13%in cat 1 and 11% in cat 4) and monocytosis (6% in cat 1 and 12.5% in cat 3). All of the present cats were treated with oxytetracycline hydrochloride at a dose of 25 mg·kg -1 . Clinical recovery and disappearence of H. felis organisms was observed on day 29. The objective of this study was to evaluate the clinical cases of haemobartonellosis diagnosed in Ankara, with a view to establishing the clinical signs and response to treatment. To the present author’s knowledge FH has previously not been reported in Ankara, in Turkey. Feline haemobartonellosis, Turkey, clinical cases.

·iroka Z., J. Drastichova: Biochemical Markers of Aquatic Environment Contamination Cytochrome P450 in Fish. A Review. Acta Vet. Brno 2004, 73: 123-132. The paper reviews the most recent data on the role of cytochrome P450 as a biochemical marker of aquatic environment pollution. The important chemical characteristics of cytochrome P450, classification of its isoforms, its functions and involvement in the metabolism of endogenous and xenobiotic agents are described as well as methods for assessment of cytochrome P450 in biological materials. The main attention is paid to the CYP 1A isoform whose detection in fish is used in toxicological monitoring of ecosystems. Monooxygenase, EROD, pesticide, PCB, TCDD The fish, as a bioindicator species, plays an increasingly important role in the monitoring of water pollution because it responds with great sensitivity to changes in the aquatic environment. The sudden death of fish indicate heavy pollution; the effects of exposure to sublethal levels of pollutants can be measured in terms of biochemical, physiological or histological responses of the fish organism ( Mondon at al. 2001). Changes in age and species distribution in a stock fish population are general indicators of water pollution, but there are also responses specific to a single pollutant or a group of contaminants (S vobodova 1997). Biochemical markers are biochemical responses induced in the presence of a specific group of contaminants that have the same mechanism of toxic activity.

Grolichova M., P. Dvofiak, H. Musilova : Employing Ionizing Radiation to Enhance Food Safety ‐ a Review . Acta Vet. Brno 2004, 73: 143-149. Food irradiation has been employed to ensure food safety or food sterility, extend its shelf-life and reduce the losses due to sprouting and ripening or pests. In the Czech Republic, mainly spices, mixed spices and dried vegetable are exposed to ionizing radiation. The greatest suppliers of irradiated foodstuffs in Europe are Belgium, France and the Netherlands. In the USA, food irradiation is more common and there are also attempts to enforce irradiation not only for food safety, but also for technological purposes. Even though irradiation is a prospective technology, its application causes physical-chemical and biochemical changes that may affect nutritional adequacy and sensory characteristics of irradiated food. In this paper, the chemical changes of basic food components (proteins, saccharides, fats) are reviewed. The chemical changes result in radiolytic products whose risks are still subject of scientific research. It is expected that the main use of gamma irradiation will be treatment of diets for patients suffering from different disorders of the immune system, allergic patients or for the army and space flights. In the production of raw or heat non-processed foodstuffs, irradiation may be a critical control point (CCP). Gamma irradiation, food, legislative bases, radiation effects, nutritional adequacy The Purpose of Food Irradiation

Hauptman, K. , F. Tich , Z. Knotek: Clinical Diagnostics of Hepatopathies in Small Mammals: Evaluation of Importance of Individual Methods. Acta Vet. Brno 2001, 70: 297-311. Until recently the possibilities of intravital diagnosis of liver function affection in small mammals have only been utilised to a minimum extent. The necessity of early hepatic disease diagnostics in these patients is highly topical. The presented paper summarises diagnostic procedures and will show the evaluation of their practical use in small mammals. Anamnestic data evaluation, palpation of the liver and collection of fluid from the abdominal cavity for the purpose of laboratory examination play an irreplaceable role in the diagnostics of liver diseases in small mammals. Biochemical blood indices analysis includes enzymes indicating hepatocellular damage (ALT, AST, LDH), cholestasis or enzymatic activity alteration (ALP, GGT), values monitoring the liver synthesis (albumin, glucose, urea, coagulation factors, bilirubin, bile acids, ammonium). Cytology and histology of biopsy specimens make it possible to evaluate the liver state precisely. In small mammals the size of a rabbit, ferret, adult guinea pig or rat utilisable methods for the liver biopsy include: fine needle aspiration biopsy, percutaneous biopsy, biopsy under the guidance of ultrasonography, biopsy under endoscopic or otoscopic guidance and biopsy during exploratory laparotomy. In patients the size of a hamster, Djungarian hamster or mouse the indication of these methods are questionable they can casuse serious hazards to the animals life. In spite of this, modifications of liver biopsy sampling are of conclusive importance in the diagnostics of hepathopathies in small mammals. In minute small mammals we can use a modification of the percutaneous biopsy using a key-hole technique and biopsy during laparotomy. Rabbit, ferret, rodents, liver biopsy, ascites, endoscopy Veterinary medicine deals with questions of the diagnostics of hepatopathies traditionally in large farm animals as well as small domestic ones (House 1992; Carl ton and McGavin 1995). It is also the group of small mammals which are relatively frequently affected by various forms of liver disease (Meredi th and Rayment 2000). There are primary liver parenchyma affections as well as secondary hepatopathies caused by metabolic disorders, enteropathies, intoxication, nephropathies or some other systemic changes in these animals. Differentiation of primary and secondary liver parenchyma affections is very difficult and in many cases practically impossible (Dial 1995). The diagnostics of liver damage in small mammals is much more complicated than in dogs and cats. Difficulty in handling the patients is one of the reasons. They are often semi-wild and the examining surgeon is in danger of being hurt. That is why thorough examination most of small mammal patients require sedation which, however, may be an exacerbating factor. The size of the patient presents another limitation due to complications of both the basic clinical examination and the use of some diagnostic imaging methods (radiographic and ultrasound). The body weight and size also influence the volume of blood samples (it is possible to collect blood volume representing 0.5% of the body weight without major problems). It is particularly necessary to consider the financial aspect, because the cost of ACTA VET. BRNO 2001, 70: 297–311 Address for correspondence: MVDr. K. Hauptmann 1Small Animal Clinic,Faculty of Veterinary Medicine University of Veterinary and Pharmaceutical Sciences Brno Palackeho 1-3, 612 42 Brno, Czech Republic Phone: +420 5 4156 2382 Fax: +420 5 4156 2382 E-mail: hauptmank@vfu.cz http://www.vfu.cz/acta-vet/actavet.htm

Kouba V.: EPIZTEXT: The First Electronic Textbook for Education and Training in General Epizootiology. Acta Vet. Brno, 2001, 70: 211-215. First electronic textbook for undergraduate and postgraduate courses in general epizootiology was developed under the name EPIZTEXT. Its version 2.2 is operated under Microsoft WORD text processing software for WINDOWS. The English written textbook contains 34 chapters. The initial one defines the epizootiology as the science that studies animal population health and diseases, their determinants and defines methods for promotion, protection and restoration of health. The first section covers animal population characteristics of health importance, disease resistance, population health and disease measurement indicators, population structures according to epizootiological characteristics, etiological agents, their sources and ways of transmission, environmental factors, infection process, epizootic process, disease nidality, zoonoses, economic factors and consequences of population health and disease. The second section includes principles and methods for animal population health and disease investigations, information systems, analyses, monitoring and surveillance. The third section deals with principles and methods for practical actions such as decision-making about anti-epizootic strategy and measures, active creation of population health, preventive and recovery measures including disease reduction, elimination and eradication, measures against diseases common to man and animals, protection of country territory, sanitation, planning, organization and management of animal population health programmes, their results and efficiency evaluation. Learners can study perusing the text, finding easily selected terms and chapters and print parts or the whole textbook. Epizootiological methods are indispensable also for avoiding diseases spreading through international trade in animal commodities which represents serious risk of disease globalization. Population medicine, software, disease analysis, health protection, control methods, disease management Among modern tools for teaching and learning belong electronic textbooks facilitating teachers’ work and learners’ study. One of the first electronic textbooks in veterinary medicine was developed for general epizootiology education and training under the name EPIZTEXT. Epizootiology is the science which studies origin, frequency, distribution, development and extinction of animal health and disease at population level, their determinants and defines the methods for promotion, protection and restoration of health by reducing, eliminating and eradicating common diseases. Epizootiology was introduced in many veterinary faculties as undergraduate curriculum subject and in many national and international postgraduate courses. Zoonoses and their control represent a particular part of this discipline. General epizootiology includes principles and methods applicable on any specific health and disease at population level in any place and time. This discipline educates veterinarians to be able to investigate epizootiological situation and apply practical measures at field and managerial levels. An impulse to develop the first electronic textbook in epizootiology was given by “Chapters” elaborated by a group of US specialists and compiled by M. Salman and I. Gardner, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins in 1992 using WORDPERFECT 5.1 software. EPIZTEXT technical contents was based upon author’s publications (3, 4, 6) and ACTA VET. BRNO 2001, 70: 211–215 Address for correspondence: Prof. MVDr Vaclav Kouba, DrSc. P. O. B. 516 170 00 Praha 7 Czech Republic Phone: +420 2 33381088 http://www.vfu.cz/acta-vet/actavet.htm experience as teacher of undergraduate and postgraduate courses and responsible officer for disease control at local, national and international levels. Several literature sources are mentioned in References (1, 2, 7, 8, 9, 10, 11, 12,14). A syllabus of general epizootiology can be found e.g., in the Report on Consultation on Undergraduate and Postgraduate Teaching in Veterinary Public Health, World Health Organization, 1983 (13). Materials and Methods The objectives were to develop an electronic textbook as a teaching tool supporting the study of general epizootiology. The material should achieve that the learners: will know animal population characteristics of health importance, etiological agents, their sources and paths of transmission, infection and epizootic processes, influencing factors and consequences of animal population health and diseases; will be able to investigate, analyze, monitor and survey animal population health and disease situation; will be able to elaborate a strategy, identify and apply effective measures for animal population health protection and recovery (diseases reduction, elimination and eradication), investigate outbreaks, elaborate, organize, implement and evaluate animal population health programmes. Main criteria when preparing this textbook were as follows: the electronic textbook to be user-friendly; to apply procedures of a widely used word processing software package; the size of unzipped textbook to fit into one 1,44 KB diskette; to provide a wide range of information and methods for undergraduate education and postgraduate training, self-study and problem solutions; to give priority to those methods which had already proven to be useful and feasible in successful programmes of disease prevention, control, reduction and eradication and to provide methods applicable for general as well as for any specific health and diseases, including those transmissible to man. The first experimental version of general epizootiology electronic textbook was prepared in 1994 for Veterinary Faculty for Foreign Students, University of Veterinary Sciences, Ko‰ice where all subjects are taught in English. This version used WORDPERFECT 5.1. software package for IBM Personal Computers applying files merging procedures. It could be run directly from the diskette or after installation on hard disk. The text could be read from the computer screen or be printed on paper. “Table of contents” served as the menu. To get into desired chapter keys “ctrl”-“home” followed by page number were used. To facilitate conversion into newer word processing software packages all original WORDPERFECT files composing the textbook were individually included in the same diskette together with original EPIZTEXT produced in one block. Individual files could be converted separately into new word processing software and then be read or printed partly or as a whole chapter using standard operation procedures of the given text processing software. Finally, EPIZTEXT was converted into WORD 7 for WINDOWS 95 as a whole (in one file) and thus simplifying significantly the operations and opening it for future transfer into newer software versions.

Krajniaakova M., E. Bekeova, a. Lenhardt, V. Cigankova, I. Valock˘, I. Maraaek: Microscopic Analysis of the Uterine Endometrium in Postparturient Ewes. Acta Vet. Brno 1999, 68: 9-12. Earlier we found that the puerperal period of ewes lambing in February was finished by day 34 post partum.The aim of this paper was to study the puerperal changes in ewes that lambed in September. Structural changes in the endometrium of the caruncular region were studied in 12 Slovak Merino ewes that lambed in September. The animals were killed on days 7, 17, 25 and 34 post partum. Samples were taken from the caruncular region of their uterine horns. The tissue samples were fixed, dehydrated and embedded in paraffin, and 7-10 mm sections were stained with hematoxylin-eosin. Microscopic analysis was performed using a projection microscope. Simultaneously the collected material was stained with toluidine-blue, and examined on the semi-thin sections. The epithelium above the caruncles was entirely destroyed on day 7. Damaged mitochondria and dilated cisterns of the endoplasmic reticulum were found in the electron micrographs. On day 17, the epithelium covered gradually a sizeable caruncle. The glandular epithelium was considerably degenerated. The caruncle was markedly reduced on day 25, and the endometrium was covered with epithelium. The endometrium was totally covered with epithelium, and cellular ultrastructure was no more damaged on day 34 post partum. The study of micromorphology of endometrial structure in caruncular area revealed that also in ewes that lambed in September, the puerperium is finished by day 34 post partum. Its course and timing did not differ from that seen in ewes lambing in February. The results extend the knowledge of the puerperal changes in different season of the year. Caruncular region, endometrium, ewe, histological structures, post partum, season of the year One of the factors that may unfavourably affect fertility is the course of the involutional and reparatory processes of the uterus in the postpartal period. At the onset of postpartal reproductive activity it is necessary to take into account the breed, the season of the year, nutrition, and the length of suckling (Kudlaa 1985; DoleIel 1989). The macroscopic changes occurring during involution of the sheep uterus were reported by F oote and C all (1969); Crowder et al. (1982); Botha (1976); Van Wyk et al. (1972); Krajniaakova (1990); Greyling and V an Niekerk (1991) in goat, and Massanyi (1996) in rabbits. A comprehensive histological study of the sheep uterus post partum was carried out by Uren (1935); and Van Wyk et al. (1972) who found that the involution process of the uterus was completed by day 28 post partum. B otha (1976) reported that both the season and lactation influence epithelisation of the caruncles. The author observed sheep that lambed during the out-of-mating season (August) to have epithelisation in the caruncular region finished on day 34 post partum whereas those that lambed in the lambing season (March) on day 30 post partum.

Siwicki A. K., M. Morand, M. Studnicka: III vitro Immullotoxicology - Practical Applicatioll. Acta vet. Bmo 1998.67: 303-307. The ill vitro immunology is one of the most important directions in immunotoxicology. At present. many ill vitro methods are used in control drug discovery. agents for chemotherapeutics and for monitoring of environmental contamination. In this paper, we present various methods which could by used for examination of the influence of different environmental chemicals and other xenobiotics on the immunocompetent cell activity in fish. The chemiluminescence response assay is important for study the phagocyte activity and the proliferative response assay of lymphocytes stimulated by mitogens is very important method for study the effects of xenobiotics on the lymphocyte T and B activities. We also demonstrate that ill vitro immune response assay analyzed by PFC and ELISPOT are very sensitive techniques for studies of the effects of xenobiOlics on the antibody secreting cells.

Siwicki A. K., M. Studnicka, M. Morand, F. Pozet, E. Terech-Majewska: Comparative lmmmunoroxicology a New Direction. Acta vet. Bmo 1998,67: 295-301. Increasing concentrations of xenobiotics in the environment, food and water supplies has direct or non-direct effects on the immunocompetent cell activity, cellular and humoral defence mechanisms and protection from diseases in animals and human. At this moment we must develop comparative aspects of immunotoxicology, justifying the wse of invenebrates, amphibians and especially fish and reptiles, as models. The defence mechanism indices have been used in various laboratory and field experiments to analyze the influence of xenobiotics on the defence mechanisms and disease resistance of mammals, birds, amphibians and fish. Many of the standard immunological techniques have been modified for optimal results. In this paper we present the basic immunological techniques for study of the effects of xenobiotics on the cellular and humoral defence mechanisms in fish. Xenobiotics,fish, defence mechanisms Understanding the human and animal health risk associated with environmental exposures involves defining the cascades of events between exposure to an environmental agent and the resulting effects on health. To cause health impairment, a pollutant must be absorbed into the body and result in biological change. For most environmental pollutants, little is known about. the flow of events between exposure and health effect. Biological markers are powerful tools that can be used to address many different issues confronting environmental health scientists. Effects of markers are indicators of biological responses. They provide information concerning the likely health outcomes associated with different target doses of environmental pollutants or their metabolites. Overall, biological markers offer the possibility of using human data to make health risk assessment more meaningful, realistic and host-effective. Risk assessment attempts to determine the consequence of toxicant exposure at the human population level. Biomarkers are frequently used to assess the responses and effects of toxicants on laboratory animals. Therefore, there is a need to extrapolate from animal data to human health effects, not only because most of the uncertainties associated with human epidemiological studies. The immune and host defences are highly conserved across species such that immunocompetent cells and some products of the immune system in humans, mice, rats and also in teleost fish are similar. Also, in instances where controlled human studies have been possible, results of immune function studies have been accurate predictors of effects in humans (H u g get t et al. 1989). Increasing concentrations of xenobiotics in the environment, industry, food and water Address for correspondence: A. K. Siwicki Department of Fish Pathology and Immunology IFI. 05-500 Piaseczno. Poland Phone: +-18 ~~ 756 2490 Fax: +48227562490 E-mail: irs@warman.com.pl

Schwaiger 1., R. D. Negele: PlasmaVitellogenin-aBloodParametertoEvaluateExposure of Fish to Xenoestrogens. Acta vet. Brno 1998,67: 257-264. Besides natural and synthetic estrogens, a variety of industrial chemicals and pesticides are suspected to mimic the natural estrogen l713-estradiol, thereby disrupting the animal endocrine system. Based on observations, sllch as the occurrence of hermaphroditism and feminization of male fish, many investigations focus on fish as indicator organisms for xenoestrogenic compounds within the aquatic environment. Both in vitro assays and ill vivo approaches have been developed to evaluate estrogenic effects of these toxicants. The occurrence of the female specific egg yolk precursor protein vitellogenin (Vtg) in the plasma of male fish has widely been used as an indicator or biomarker of xenoestrogen exposure. This paper briefly reviews potential xenoestrogens known so far. physiological aspects of vitellogenesis, current applications of Vtg plasma levels in fish as a biomarker for estrogenic compounds. and various aspects concerning the possible biological significance of this parameter. Xenoestrogells. \'itellogenin. fish

Svobodova Z., L. Groch, M. Flajshans, B. Vykusova. 1. Machova: The Effect of Long-term Therapeutic Bath of Malachite Gteen on Common Carp (C-,prinus carpio L.). Acta vet. Bmo 1997,66: 111-117. The aim of this paper was to determine acute toxicity of malachite green of the Vert Malachite BE SA Crystaux type for common carp and to evaluate the effect of a 6-day therapeutic bath in O.S mg.!-I concentration of this pharmaceutical on values of selected haematological indicators including micronuclei test, as well as on histological composition of tissues of oneto two-year-old carp. The 48hLCSO value of the malachite green tested was 0.84 mg.!-I. After the 6-day bath, changes in red blood picture [decrease in haematocrit value PCV (P

Kala~njuk G. I., M. Marounek, L.G.Kala~njuk, M. G. Gerasymiv, O. G. S a v k a : Activity of Dehydrogenases and Enzymes of Nitrogen Metabolism in Cardiac Tissue and Skeletal Muscle of Steers fed Monensin. Acta vet. Brno 1995, 64: 157-161. Activites of lactate dehydrogenase (E.C. 1.1.1.27), malate dehydrogenase (E.C. 1. 1. 1.40), 2-oxoglutarate dehydrogenase, (E.C.1.2.4.2) glutamate dehydrogenase (E.C.1.4.1.3), glutamin synthetase (E.C.6.3.1.2), arginase (E.C.3.5.3.1), ornithine carbamoyltransferase (E.C.2.1.3.3), aspartate aminotransferase (E.C.2.6.1.1) and alanine aminotransferase (E.C.2.6.1.2) were measured in cardic and skeletal muscles of steers fed ration with or without monensin. Steers, 9 months old at the beginning, were fed concentrate, molasses, grass, lucerne and maize chaff. Five steers received monensin in amount of 0.5 mg per 1 kg of live body mass per day. Control ration, without monensin was fed to the other five steers. After 10 months steers were slaughtered and activity of enzymes assayed in the mitochondrial and cytoplasmic fraction of cells. Monensin increased activity of 2-oxoglutarate dehydrogenase and decreased activity of glutamate dehydrogenase in both fractions of the skeletal muscle tissue. Enzymatic activities found in heart mitochondria were higher in monensin-fed steers than in control steers. Steers given monensin gained 8.23 % more than control steers (263 vs 243 kg). Monensin, enzyme, heart, muscle Feed antibiotics, including monensin, have the ability to improve perfonnance and feed efficiency in ruminants. It is known that monensin is absorbed from the alimentary tract (D a vis 0 n 1983; Don 0 h 01984) and several authors demonstrated its influence on intermediary metabolism of ruminants (Armstrong and Spears 1988; Benz et al.1989;Marounek et al. 1989). Little is known about the effect of monensin on activity of tissue enzymes in ruminants. K a I a c n j uk et al. (1993) measured activity of various enzymes in rumen mucosa and liver of steers fed monensin at recommended level. Authors found no consistent effect of monensin on enzymes of both tissues. In this paper we report data on activity of nine enzymes in cardiac and skeletal muscles of steers fed ration with or without monensin for ten months. Cardiac and skeletal muscles are primary target tissues at high intake of monensin (T 0 d d et al. 1984; Van V lee t et al. 1985) and, presumably, also during long-term supplementation of rations under normal feeding conditions. ' Materials and Methods Ten crossbred steers, 9 months old at the beginning of experiment, were divided into two groups, according to feed additive treatment. Steers were individually housed and kept on a diet consisting of concentrate (I kg per 100 kg of the live weight), molasses (0.5 1.0 kg) and grass, lucerne and maize chaff ad libitum. In winter, the roughage portion of the diet consisted of maize silage and beet ad libitum. Concentrate contained ground barley (63%), dried poultry litter from broilers which did not receive a ionophore in their diet (20%), grass meal (15%) and zeolite (2%).

Skrzypczak W. F., K. Janus, Z. Muszczynski and D. Jankowiak: The effect of spironolactone (Aldactone) on electrolyte balance and renolfuncions of calves. Acta vet. Bmo, 63, 1994: 19-23. A short spironolactone test was carried out in a group of 10 clinically healthy calves at the age of 4 weeks. For 5 days (every 24 hours) the calves were administered Aldactone (Searle), per os, at a dose of 1 mglkg of body weight. Before the test began and 120 and 144 hours after giving the first spironolactone dose, the effective renal blood (ERBF) and plasma (ERPF) flow, and the glomerular filtration rate (GFR) were measured using sodium salt of p-aminohippuric acid and inulin. The levels of sodium, potassium and chlorides in plasma and erythrocytes were also determined. Some changes in the concentration of tested electrolytes were found in both the plasma and the intracellular fluid of erythrocytes. Changes in the renal function of calves were also indicated by an increase in effective renal plasma flow as well as by a decrease in glomerular filtration rate. Spironolactone, electrolyte b(llance, renal.fi!,nction, calves Calf kidneys are functionally efficient in the fIrst month oflife. However, they reveal some features of functional immaturity. These involve lowered effective renal blood (ERBF) and renal plasma (ERPF) flow,low glomerularfiltrationrate and low ability of urine concentration (Mercer etal.1978; Wanner etal.1981; Hartmann et al. 1987; Skrzypczak 1989; Skrzypczak etal. 1989; Skrzypczak 1991). The observed high tabular resorption ofNa, K and Cl in the frrst ~ days of life (Safwate et al., 1980; Skrzypczak, 1991) becomes lower as the age of calves increases (Safwate etal.1982; Skrzypczak 1989). The results of experiments show that, by the third day of life, mechanisms which control sodium resorption become active and kidney tubules respond to aldosterone infusion with the growth ofNa resorption (Safwate et al. 1980; Safwate et al. 1982; Itoh et al. 1985; Safwate 1985). Some authors, however, found a lack of correlation between the plasma renin activity and the aldosterone level, and sodium and potassium concentrations in the blood plasma (Siegler etal.1977; Moncaup etal.1980; Safwate etal.1982). Cabello (1980)evenreportsanegative correlation between plasma levels of aldosterone and sodium. Jankowiak (1989) showed that administering DOCA at a dose of 0.1 mglkg body weight Lm. caused a clearance drop and the urine excretion of sodium, potassium and chlorides also decreased. Safwate (1985) also noticed a lowering of sodium excretion in urine after aldosterone infusion, although he did not observe any changes in potassium excretion. Some authors indicate that there is a possibility of metabolic stimulation of the calf suprarenal glands in the first month of life (McEwan et al. 1968; Baranow-Baranowski etal. 1987). As can be seen from the literature review, there are many issues connected with tubular activity regulation in the postnatal period pertaining to electrolytes resorption which have not been fully explained yet (K a g a w a 1960; Bra u n Ii chand K e r s ten 1972; Cor v 0 I et al. 1981). The objective of this paper was to examine the effect of spironolactone (Aldactone) on electrolyte balance of body fluids and on glomerular filtration rate and effective blood and plasma flow through young calf kidneys. Materials and Methods The experiment was carried out on a group of 10 clinically healthy calves of cb. breed at the age of 4 weeks. They were kept in a calf-pen under constant, optimal conditions. Their left and right jugular vein had been catheterized and they had been weighed before the experiment began. Prior to the treatment period, the effective renal plasma (ERPF) and blood (ERBF) flow and the glomerular filtration rate (GFR) were defined. The levels of sodium, potassium and chlorides in blood plasma and erythrocytes were examined. To assess ERPF and GFR, sodium salt of p-aminohippuric acid (Serva-GmbH) and inulin (Sigma Chern. Co.) were used respectively. The clearance of these substances was measured using Bettge' s method. On the basis of ERPF and haematocrit, the amount of ERBF was defined and the filtration fraction (FF) was calculated. In order to block aldosterone activity, a short spironolactone test was conducted. The calves were administered 1 mglkg body weight of Aldactone (Searle) per os for 5 days. After 120 and 144 hours from giving the first dose of Aldactone, new examinations of ERPF, GFR values and Na, K and Cllevels in plasma and whole blood were carried out.

The paper describes the first occurrence of myelocytomatosis in the Czech Republic. A group of hens from the breeding flocks of the meat type aged 27 and 50 weeks with increased mortality and tumorous disease occurrence was examined. Necropsy and pathohistology proved myelocytomatosis. The presence of the ALV subtype J virus was proved using PCR.

The differences are described in this paper as observed in the formation of antibodies an adult hens following application of different routes of immunization and using two antigens distinguished by different dependences on the thymus, actually the SRBC and B. abortus. Formation of the agglutination antibodies to SRBC was inferior to the latter, the i.m. immunization method of antigen application with adjuvant, in conjuction with simplicity of the procedure, in general leading to optimum titres of specific antibodies.

SimunekJ., A. B. Siddique, E. Hegerova: Placental Transfer of Suljamethazine in Mice and Rats. Acta Vet. Brno, 47,1978: 145-151. Placental transfer of sulfamethazine in mice was investigated in the second half of their pregnancy. The transfer was not free but only a part of the drug administered to mothers was found to pass from the maternal circulation to the fetus. Placental transfer of sulfamethazine in rats was investigated at two stages of ge­ station - early (8th to 10th day) and late (18th to 20th day of gestation). Ther~ was free and rapid passage of the drug from the maternal circulation to the fetus at the early stage of gestation. But the passage was found to be greatly reduced at the late stage of gestation. The drug also appeared in the amniotic fluid. Mice, rat, placenta, fetus, transfer, suljamethazine~· The placenta in unique is its function. Its life time is short relative to the fetus, and its size and function change continually during the course of gestation (Wynn 1968). The study of the placental transfer of drugs is of paramount importance because much of intrauterine fetal welfare depends on the exchange processes of these substances through the placenta. Sulfonamides have been reported to easily pass through both human and animal placenta (Lee, Anderson and Chen 1938; Friesen 1951; Siddique and Simunek 1977). The possible use of these drugs for intrauterine therapy as well as for fetal prophylaxis and their effects on the fetus and newborn make it necessary to know and understand the degree. and extent of transplacental passage of these drugs. The present paper reports on the placental transfer of sulfamethazine in mice in their second half of pregnancy and in rats at two stages of pregnancy.

Vlcek Z.: LatePuerperium and the New Reproductive Cycle in Cows with Regard to Restoration of their Ovarial Activity, Estrus and the First Postpartal Ovulation. Acta vet. Brno, 51, 1982: 75-80. The time sequence of uterine involution, regression of the corpus luteum gravidi­ tatis, restoration of the ovarial activity and symptoms of estrus and the first postpartal ovulation were followed in 282 Bohemian Pied cows. The involution of the reproductive organs inspected by external, vaginal and rectal examination followed the usual pattern. More pregnancies were placed in the right uterine horn. A pea-sized regressive corpus luteum graviditatis was found in a portion of examined cows on days 12-14 post partum, later no more corpora lutea were exactly identifiable. In several cows as soon as on days 15 -17 post partum, in majority of animals on days 18-20, growth of follicles was observed. The majority of cows showed signs of the jollicular phase of the reproductive cycle in the fourth week after parturition and some animals were in metestrus or die­ strus. Morphological changes in the first cycle were more frequently observed on the left ovary and on the side of the previously non-pregnant uterine horn. Involution, uterus, ovary, vaginal mucosa, secretions. Great attention has been paid to physiology of puerperium in cows as well as to etiology, diag­ nosis and prevention of pathological conditions complicating the course of puerperium and im­ pairing future fertility. The present study reports on results of a complex examination of puerpe­ ral cows obtained under large-scale farm conditions. The aim of the research project is to suc­ cessfully monitor the reproductive process in cattle. New data have been added to the knowledge of the course of puerperium in cows, including information about mechanisms of puerperal involution of reproductive organs, regression of the corpus luteum graviditatis, restoration of the reproductive cycle after parturition (Wagner et a!. 1969; Wagner and Hansel 1969; Kudl:ic and Vlcek 1975; Kudl:ic 1975; LaktU; 1979). The course of involutional process was studied in detail by Gier and Marion (1968), factors affect­ ing the regression of c. luteum by Marion et a1. (1968), the bacterial content of the uterus by Lloyd et a1. (1968). Regression of corpus luteum graviditatis was investigated also by Wohanka (1961) and others. Uterine involution and follicular activity of the ovaries in puerperium with re­ gard to fertility disorders was studied by Bostedt et a1. (1976), Arbeiter (1976), Buchholz and Busch (1977), Rtisse (1968). Relationships between the course of puerperium and readiness for a new conception was followed by Bach (1974). The relationship between the course of puer­ perium, milk yield, lactation curve and conception rate was investigated by Buchholz and J e­ richow (1977). The aim of these and many other papers was to optimalize and shorten the course of puerperium and to ensure a high conception rate in cows through an active control of puerperium and, if necessary, by means of therapy.