Infection dynamics following experimental challenge of pigs orally dosed with different stages of two archetypal genotypes of Toxoplasma gondii.

Toxoplasma gondii is a food-borne zoonotic parasite widespread in a variety of hosts, including humans. With a majority of infections in Europe estimated to be meat-borne, pork, as one of the most consumed meats worldwide, represents a potential risk for consumers. Therefore, we aimed to investigate the progress of T. gondii infection and tissue tropism in experimentally infected pigs, using different T. gondii isolates and infectious stages, i.e. tissue cysts or oocysts. Twenty-four pigs were allocated to treatment in four groups of six, with each group inoculated orally with an estimated low dose of either 400 oocysts or 10 tissue cysts of two European T. gondii isolates, a type II and a type III isolate. The majority of pigs seroconverted two weeks post-inoculation. Pigs infected with the type III isolate had significantly higher levels of anti-T. gondii antibodies compared to those infected with the type II isolate. Histopathological exams revealed reactive hyperplasia of the lymphatic tissue of all pigs. Additionally, a selected set of nine tissues was collected during necropsy at 50 dpi from each of the remaining 22 pigs for T. gondii DNA detection by quantitative real-time PCR. A positive result was obtained in 29.8 % (59/139) of tested tissues. The brain was identified as the most frequently positive tissue in 63.6 % (14/22) of the animals. In contrast, liver samples tested negative in all animals. The highest mean parasite load, calculated by interpolating the average Cq values on the standard curve made of ten-fold serial dilutions of the genomic DNA, corresponding to 100 to 104 tachyzoites/µL, was observed in shoulder musculature with an estimated concentration of 84.4 [0.0-442.5] parasites per gram of tissue. The study highlights the variability in clinical signs and tissue distribution of T. gondii in pigs based on the combination of parasite stages and strains, with type III isolates, particularly oocysts, causing a stronger antibody response and higher tissue parasite burden. These findings suggest the need for further investigation of type III isolates to better understand their potential risks to humans.

Reproductive tract morphology and symmetry of farmed common eland (Tragelaphus oryx) bulls, and their relationship with secondary sexual traits and social rank.

Common eland bulls are important game ranching animals in southern Africa, for tourism, breeding/live sales, trophy hunting, and game meat production. Interest has grown in their production potential, intensifying animal husbandry and breeding approaches. However, little scientific information is available regarding the intensive management of this species, including information regarding scientifically based selection criteria for breeding bulls. Often, simple phenotypic traits are utilised unless high-value game animals are being bred, with horn and body size often being the only traits considered for breeding bulls. This study provides basic information about the reproductive tract morphology, symmetry, and their relationships with secondary sexual traits, social rank, and temperament in common eland. Six eland males (2-2.5 years old; 203 ± 20 kg) were utilised for the study. Social rank, body weight and body condition score, temperament score, and horn size were determined before the animals were culled, and their reproductive tracts were collected for morphometric and histological evaluation. Results indicate relatively low asymmetry in bilateral reproductive tract components. Individual traits of eland bulls such as age, body weight, and social rank correlated the development of some testicular morphologies and thus may possibly be used in zoos, game ranching, and commercial production for the selection of breeding males, but not temperament and horn length, which should be carefully evaluated. Whilst this study provides baseline information from a limited sample size, further studies should incorporate the assessment of testicular hemodynamics, as well as the effects of factors such as season and androgen secretion patterns over a wider age range of common eland males and populations to provide further information for the management practices of breeding common eland bulls.

Effect of GnRH agonist (deslorelin) on reproductive activity in captive female veiled chameleons (Chamaeleo calyptratus).

Eighteen 5 months old veiled chameleon females (Chamaeleo calyptratus) were used in the study. Seven females received subcutaneous implants with gonadotropin-releasing hormone agonist (GnRH) deslorelin acetate whereas eleven females were used as control animals without any implants. Females were kept in five terraria, in groups of four females (in 3 terraria) and groups of three females (in 2 terraria), respectively. A minimum of one female with GnRH implants was present in each terrarium. They were kept under standard husbandry conditions. Females of both groups (females with GnRH and controls, respectively) were monitored for three years. No differences between females with GnRH implants and females without GnRH implants were observed with respect to the presence of large ovarian follicles, number of eggs/female, or number of clutches/female. GnRH implants did not prevent spontaneous reproductive activity in any of the 7 females with implants. Ovariectomy was performed in 8 females (4 females with GnRH implants and 4 control females). Ten females (3 females with GnRH implants and 7 control females) had to be euthanised. In 17 of 18 female veiled chameleons of this study histologic examination revealed heterophilic granulomatous oophoritis. The use of GnRH agonist implants did not prove to be an appropriate method for the control of reproductive function in captive female veiled chameleons.

Diagnosis of Aspergillosis in Horses.

Invasive pulmonary aspergillosis (IPA) may be a rare cause of granulomatous pneumonia in horses. The mortality of IPA is almost 100%; direct diagnostic tools in horses are needed. Bronchoalveolar lavage fluid (BALF) and serum samples were collected from 18 horses, including individuals suffering from IPA (n = 1), equine asthma (EA, n = 12), and 5 healthy controls. Serum samples were collected from another 6 healthy controls. Samples of BALF (n = 18) were analyzed for Aspergillus spp. DNA, fungal galactomannan (GM), ferricrocin (Fc), triacetylfusarinin C (TafC), and gliotoxin (Gtx). Analysis of 24 serum samples for (1,3)-ß-D-glucan (BDG) and GM was performed. Median serum BDG levels were 131 pg/mL in controls and 1142 pg/mL in IPA. Similar trends were observed in BALF samples for GM (Area under the Curve (AUC) = 0.941) and DNA (AUC = 0.941). The fungal secondary metabolite Gtx was detected in IPA BALF and lung tissue samples (86 ng/mL and 2.17 ng/mg, AUC = 1).

Is a new generation of mycotoxin clay adsorbents safe in a pig's diet?

BACKGROUND: Bentonites, as a clay mineral, serve in pig farms as adsorbents of toxic substances. They are mainly used to reduce the negative impact of mycotoxins to maintain the performance and health status of animals. The new genotypes of pigs are highly sensitive to a range of antinutrients, including mycotoxins. Currently, attention is focused on more effective adsorbents of mycotoxins with a higher adsorption capacity. Such materials are in great demand among feed manufacturers. However, there is a concern that these new materials may also adsorb too many essential nutrients and decrease animal performance. The aim of the experiment was to evaluate the effect of the new generation of purified bentonites on the efficiency and health status of the pigs. RESULTS: Forty-eight slaughtered pigs with an average weight of 31.2 ± 2.6 kg were included in the experiment. The pigs were divided into two groups (2 × 24 pigs). Pigs were slaughtered at an average weight of 66.3 ± 5.2. The first group had a diet without clay (control-C). The second group (treatment-T) was fed a diet with a clay additive (purified bentonite) of 1.5 kg/t. Animals were fed the experimental diet for 35 days. In group T, a higher daily weight gain (by 4.8%) and feed intake (by 2.9%) was observed while the feed conversion decreased by 1.9%. There were no significant differences between the groups of pigs during observation in the evaluation of hematological, biochemical parameters of the blood. Morpho-pathological analysis of the jejunum showed similar signs of moderate lymphoplasmacytic infiltrate in the mucosa in the groups C and T, contained similar number of goblet cells. CONCLUSION: Taken together, the addition of the new generation of bentonite clays did not negatively influence the health status and the performance of pigs.

Antibiotic-Loaded Amphiphilic Chitosan Nanoparticles Target Macrophages and Kill an Intracellular Pathogen.

In this work, levofloxacin (LVX), a third-generation fluoroquinolone antibiotic, is encapsulated within amphiphilic polymeric nanoparticles of a chitosan-g-poly(methyl methacrylate) produced by self-assembly and physically stabilized by ionotropic crosslinking with sodium tripolyphosphate. Non-crosslinked nanoparticles display a size of 29 nm and a zeta-potential of +36 mV, while the crosslinked counterparts display 45 nm and +24 mV, respectively. The cell compatibility, uptake, and intracellular trafficking are characterized in the murine alveolar macrophage cell line MH-S and the human bronchial epithelial cell line BEAS-2B in vitro. Internalization events are detected after 10 min and the uptake is inhibited by several endocytosis inhibitors, indicating the involvement of complex endocytic pathways. In addition, the nanoparticles are detected in the lysosomal compartment. Then, the antibacterial efficacy of LVX-loaded nanoformulations (50% w/w drug content) is assessed in MH-S and BEAS-2B cells infected with Staphylococcus aureus and the bacterial burden is decreased by 49% and 46%, respectively. In contrast, free LVX leads to a decrease of 8% and 5%, respectively, in the same infected cell lines. Finally, intravenous injection to a zebrafish larval model shows that the nanoparticles accumulate in macrophages and endothelium and demonstrate the promise of these amphiphilic nanoparticles to target intracellular infections.

Nanocrystalline chloroxine possesses broad-spectrum antimicrobial activities and excellent skin tolerability in mice.

Background: Antimicrobial submicrometer particles are being studied as promising interventions against a wide range of skin conditions, such as fungal or bacterial infections. Aims: To submicronize chloroxine, the crystalline compound 5,7-dichloro-8-hydroxyquinoline, by nanoprecipitation and characterize the resulting assemblies. Methods: The chloroxine particles were stabilized by a nonionic surfactant and were studied by a broth microdilution assay against 20 medically important bacteria and fungi. The intervention was studied using a murine model of skin irritation. Results & conclusion: Chloroxine nanoparticles with a diameter of 600-800 nm exhibit good tolerability in terms of skin irritation in vivo and good antimicrobial activity. Thus, the fabricated formulation shows great promise for interventions for both cutaneous infection control and prophylaxis.

Case Report: Spontaneous Appendicitis With Suspected Involvement of Klebsiella variicola in Two Pet Rabbits.

Although laboratory rabbits are commonly used as models of appendicitis in man, spontaneous appendicitis was only described ante-mortem in one pet rabbit with an acute abdomen. The aim of this article is to describe two spontaneous cases of appendicitis in pet rabbits, to describe therapeutic appendectomy, and to discuss the microbial flora of the inflamed appendix. A 5-month-old intact female and a 16-month-old, neutered male were presented to the veterinary clinic with restlessness, anorexia, and reduced faecal output. The main clinical findings were restlessness, severe discomfort on abdominal palpation, a mid-abdominal palpable tubulous mass and an elevated rectal temperature. Blood analyses showed lymphocytosis, monocytosis, and hyperglycaemia. Radiography was inconclusive. Abdominal ultrasound revealed a presence of a tubular structure with wall thicknesses of 4.2 and 3.7 mm in the two rabbits, respectively. The tubular structure had a rounded, closed end, and a multilayered wall, suggestive of appendicitis. Due to metabolic acidosis and poor prognosis, the first rabbit was euthanized. In the 16-month-old rabbit, appendectomy was performed. Recovery was uneventful, and 4 h after surgery, the rabbit started to become normally active. Postoperative care consisted of fluid therapy, multimodal analgesia, supportive care and prokinetics. Follow-up examinations at 10 days, 1 month, and at 11 months after the surgery did not show any abnormal clinical or laboratory findings. Histopathological examination of appendices from both rabbits showed gangrenous appendicitis. Aerobic cultivation showed the presence of pure culture of Klebsiella variicola sensitive to enrofloxacin, marbofloxacin, tetracycline, cefuroxime, trimethoprim sulphonamide, neomycin, and gentamicin. Restlessness associated with anorexia, abdominal pain, palpable abdominal mass, hyperglycaemia, lymphocytosis, and elevated rectal temperature may be indicative of inflammation within the gastrointestinal tract. Abdominal ultrasound is recommended in rabbits with showing these clinical signs because radiography can be inconclusive. Appendicitis is a life-threatening condition, which should be included into the list of differential diagnoses; for the rabbit, an acute abdomen and gastrointestinal stasis syndrome and must be treated immediately. K. variicola may be associated with appendicitis in rabbits as a causative agent or in association with appendix intraluminal dysmicrobia.

Protective effect of a new generation of activated and purified bentonite in combination with yeast and phytogenic substances on mycotoxin challenge in pigs.

The study aimed to investigate the efficacy of new mycotoxin adsorbents based on purified and activated bentonites combined with yeast and phytogenic compounds in fattening pigs. The experiment involved 96 pigs (31.2±2.4 kg). Control (C) group was fed a diet naturally contaminated with mycotoxins (5 mg/kg deoxynivalenol, DON) without an adsorbent. Treated groups received the feed with mycotoxin adsorbents: purified and activated bentonite (T1), purified and activated bentonite, yeast derivatives, phytogenic substances (T2), and purified, activated, and sulphurated bentonite with phytogenic substances (T3). Evaluated parameters involved growth performance, organ weight, small intestine and liver histopathology, complete blood count, serum biochemistry, antioxidant status of the organism and total and free DON content in urine. In all treated groups, an significant increase in intestinal GSH and GSH/GSSG ratio was observed when compared to C. No significant effects on liver and kidney weight, complete blood count, serum or intestinal malondialdehyde concentration, or total/free DON content in urine were observed. All adsorbents improved histopathological findings in the liver when compared to C. Moreover, T1, and T2 groups showed no presence of inflammatory reaction or necrotic changes in the livers. Although, mycotoxin adsorbents investigated in this study had no significant impact on pig growth performance, they reduced the oxidative stress, and on the tissue level they protected the jejunal tissue and liver parenchyma under deoxynivalenol challenge.

Penicillin Treatment Failure in Rabbit Syphilis Due to the Persistence of Treponemes (Treponema paraluisleporidarum Ecovar Cuniculus) in the Focus of Infection.

Rabbit venereal spirochetosis, a disease caused by Treponema paraluisleporidarum ecovar Cuniculus (TPeC), affects both wild and pet rabbits, and is transmitted sexually and via direct contact among animals. Treatment of syphilis in pet rabbits requires administration of antibiotics, including penicillin G, chloramphenicol, or fluoroquinolones. The aim of this work was to elucidate the cause of penicillin treatment failure in rabbit syphilis in a pet rabbit treated in Brno, Czech Republic, and to assess the phylogenetic relatedness of the agent to previously characterized pathogenic treponemes. Following amputation of the infected digits, the second round of penicillin treatment using the same dosage and application route resulted in the disappearance of clinical symptoms within a period of two weeks. The bacterium was successfully isolated from the claws, propagated in three experimental rabbits, and the resulting TPeC strain was designated as Cz-2020. Analysis of four genetic loci revealed that the Cz-2020 strain was similar but also clearly distinct from the only TPeC strain, which had been characterized in detail to date, i.e., the Cuniculi A strain, which was isolated in North America. The strain Cz-2020 represents the first available viable TPeC strain of European origin. DNA sequences encoding five penicillin-binding proteins of the strain Cz-2020 were compared to those of Cuniculi A, which is known to be sensitive to penicillin. The sequences differed in six nucleotides resulting in single amino acid changes in Penicillin-binding protein 1, 2, and 3. Since the second round of treatment was successful, we conclude that the penicillin treatment failure in the first round resulted from the presence of infection foci in claws where treponemes persisted.

The Expression of Selected Wnt Pathway Members (FZD6, AXIN2 and ß-Catenin) in Canine Oral Squamous Cell Carcinoma and Acanthomatous Ameloblastoma.

The Wnt signaling pathway is well known to be involved in many types of human cancer; however, in veterinary medicine, the investigation of individual Wnt members' expression, and their role in or association with oral tumor pathogenesis, is still underevaluated. We aim to determine the expression pattern of Frizzled-6 (FZD6) as one of the Wnt receptors in two of the most common canine oral neoplastic lesions-canine oral squamous cell carcinoma (COSCC) and canine acanthomatous ameloblastoma (CAA). While COSCC is a malignant tumor with aggressive biological behavior and a tendency to metastasize, CAA is a benign tumor with high local invasiveness. In CAA, the expression of FZD6 was mostly located in the center of the epithelial tumorous tissue, and cells exhibiting features of squamous metaplasia were strongly positive. In well-differentiated COSCC, FZD6 was expressed in the tumorous epithelium as well as the tumorous stroma. There was a negative correlation between cytokeratin expression and FZD6 expression in COSCC, where the central parts of the epithelial tumorous tissue were often FZD6-negative. The non-differentiated COSCC with low expression of cytokeratin exhibited a diffuse FZD6 signal. The invasive front with areas of tumor budding exhibited high FZD6 expression with a loss of cytokeratin expression. Moreover, the expression of ß-catenin and AXIN2 was increased in comparison to gingiva. In conclusion, our study revealed significant differences in the expression patterns and the levels of FZD6 between COSCC and CAA, indicating the differential engagement of the Wnt pathway in these tumors.

Glyphosate Interaction with eEF1α1 Indicates Altered Protein Synthesis: Evidence for Reduced Spermatogenesis and Cytostatic Effect.

The broad-spectrum herbicide, glyphosate, is considered safe for animals because it selectively affects the shikimate pathway that is specific to plants and microorganisms. We sought a previously unknown mechanism to explain the concerns that glyphosate exposure can negatively affect animals, including humans. Computer modeling showed a probable interaction between glyphosate and eukaryotic translation elongation factor 1 subunit alpha 1 (eEF1α1), which was confirmed by microcalorimetry. Only restricted, nondisrupted spermatogenesis in rats was observed after chronic glyphosate treatments (0.7 and 7 mg/L). Cytostatic and antiproliferative effects of glyphosate in GC-1 and SUP-B15 cells were indicated. Meta-analysis of public health data suggested a possible effect of glyphosate use on sperm count. The in silico, in vitro, and in vivo experimental results as well as the metastatistics indicate side effects of chronic glyphosate exposure. Together, these findings indicate that glyphosate delays protein synthesis through an interaction with eEF1α1, thereby suppressing spermatogenesis and cell growth.

Antitubercular nanocarrier monotherapy: Study of In Vivo efficacy and pharmacokinetics for rifampicin.

Tuberculosis represents a major global health problem for which improved approaches are needed to shorten the course of treatment and to combat the emergence of resistant strains. The development of effective and safe nanobead-based interventions can be particularly relevant for increasing the concentrations of antitubercular agents within the infected site and reducing the concentrations in the general circulation, thereby avoiding off-target toxic effects. In this work, rifampicin, a first-line antitubercular agent, was encapsulated into biocompatible and biodegradable polyester-based nanoparticles. In a well-established BALB/c mouse model of pulmonary tuberculosis, the nanoparticles provided improved pharmacokinetics and pharmacodynamics. The nanoparticles were well tolerated and much more efficient than an equivalent amount of free rifampicin.

Comprehensive fitness evaluation of common carp (Cyprinus carpio L.) after twelve weeks of atrazine exposure.

Atrazine is frequently detected in surface waters and negatively affect physiological function in aquatic organisms. Even though numerous authors have intensively studied its toxicity, only limited information concerning the long-term fish exposure is available. The aim of this study was evaluation of chronic exposure in common carp. Fish were exposed to a range of atrazine concentrations (0.3 - environmentally relevant concentration; 300; 1000; and 3000 µg/l) for twelve weeks. The potential impact of atrazine exposure was studied using various markers (behaviour; biometrical characteristics; haematological, biochemical and oxidative stress indices and histopathological changes). Most alterations were recorded at the highest concentration (3000 µg/l) which is probably due to a combined effect of both the herbicide exposure and intensive parasite infection development during second week. This group was cancelled after three weeks due to adverse health status, which partly confirmed atrazine immunotoxicity. Chronic exposure resulted in long-term reduction in feed intake followed by a significant decrease in body weight and morphological changes in gill at 1000 µg/l. At the same concentration, significant alterations in haematological (e.g. increase in erythrocyte, leukocyte, lymphocytes and neutrophil counts as well as decrease in monocyte counts) and biochemical (e.g. changes in enzyme activities; increase in glucose; decrease in ammonia) indices were documented, especially during first three weeks. Similar trend, but not so intensive, was observed at 300 µg/l. Moreover, significant changes were observed in various indicators of oxidative stress. These alterations were highly variable with both increasing and decreasing trends depending on dose and analysed tissue. Significant changes, especially in white blood profile, enzyme activities and oxidative stress indices, were proven even at 0.3 µg/l. Obtained results indicate that chronic atrazine exposure of common carp can negatively influence many indicators of health status such as behaviour, immune response, haematological and biochemical profile, oxidative stress indices and organ histopathology.

The effects of ciprofloxacin on early life stages of common carp (Cyprinus carpio).

The authors performed a toxicity test with ciprofloxacin in fertilized eggs of common carp according to guideline 210 of the Organisation for Economic Co-operation and Development. The tested concentrations were 1 µg L(-1) , 100 µg L(-1) , 500 µg L(-1) , 1000 µg L(-1) , and 3000 µg L(-1) . Accelerated hatching was found in all groups exposed to ciprofloxacin, but significant growth reduction was found only in the group exposed to the highest concentration (3000 µg L(-1) ). Increased numbers of macroscopic morphological anomalies were observed on day 6 of the test (after hatching). The highest numbers of macroscopic morphological anomalies were observed in the groups of free embryos and larvae exposed to ciprofloxacin concentrations of 100 µg L(-1) , 500 µg L(-1) , 1000 µg L(-1) , and 3000 µg L(-1) (20-23% of tested samples). A gradual decrease in glutathione S-transferase activity was detected in all experimental groups exposed to ciprofloxacin, but significant differences (p < 0.01) were found only in groups treated with 500 µg L(-1) and 3000 µg L(-1) . Glutathione peroxidase and catalase exhibited increased activity in most of the tested concentrations (p < 0.01 and <0.05, respectively), whereas decreased glutathione reductase activity was found in the groups exposed to ciprofloxacin concentrations of 500 µg L(-1) and 3000 µg L(-1) (p < 0.05). The concentration of thiobarbituric acid-reactive substances was significantly lower (p < 0.01) in all experimental groups exposed to ciprofloxacin. The lowest-observed-effect concentration of ciprofloxacin was 1 µg L(-1) . These results suggest that hatching, early ontogeny, occurrence of morphological anomalies, antioxidant and biotransformation enzyme activity, and lipid peroxidation in fish can be affected by ciprofloxacin. Environ Toxicol Chem 2016;35:1733-1740. © 2015 SETAC.

Lethal Encephalitozoon cuniculi genotype III infection in Steppe lemmings (Lagurus lagurus).

Microsporidia are ubiquitous, spore-forming, intracellular parasites infecting invertebrates and vertebrates. Some of them are important opportunistic pathogens in humans, including three species of genus Encephalitozoon. Intraspecies genetic variation with a different range of hosts is known in Encephalitozoon cuniculi distinguishing four genotypes. Recently, E. cuniculi is often observed in pet animals, mainly E. cuniculi genotype I in pet rabbits. This study described a fatal encephalitozoonosis in a group of pet rodents Steppe lemmings (Lagurus lagurus). The animals were presented with progressive weight loss, aggression, cannibalism, purulent conjunctivitis and hind limb paresis. Death occurred within 48 h after the onset of clinical signs. The group comprised of 15 animals was affected and died within a period of three months. Post-mortal examination did not show any macroscopic changes. Microsporidial vacuoles with typical spores were found in brain and kidney tissues and E. cuniculi DNA in all tested organs. The internal transcribed spacer region (ITS) of rRNA gene showed 100% homology with E. cuniculi genotype III previously identified in dogs, tamarin colonies from zoos, swine, birds and humans. Pet lemmings could represent a new potential source of the infection for their breeders.

Evaluation of biochemical, haematological, and histopathological responses and recovery ability of common carp (Cyprinus carpio L.) after acute exposure to atrazine herbicide.

The aim of study was to evaluate the effect of atrazine exposure (5, 15, 20, and 30 mg·L(-1)) on common carp and the ability of regeneration. During 96 h exposure we observed abnormal behavior in fish exposed to 20 and 30 mg·L(-1). Mortality and histological alterations were noticed only in the group exposed to 30 mg·L(-1). Most experimental groups showed significantly (P < 0.05) lower values of haemoglobin, haematocrit, leukocyte, and lymphocyte and significantly higher values of monocytes, segmented and band neutrophile granulocytes, and also metamyelocytes and myelocytes. A significantly lower (P < 0.05) leukocyte count was also recorded in experimental groups (5 and 15 mg·L(-1)) after recovery period. Statistically significant (P < 0.05) alterations in glucose, total protein, lactate, phosphorus, calcium, and biopterin as well as in activities of ALT, AST, ALP, and LDH were found in most experimental groups. These changes were most apparent in the groups exposed to 20 and 30 mg·L(-1). Most of the indices were found to be restored after the 7-day recovery period with the exception of LDH, ALT, and lactate in the group exposed to 15 mg·L(-1). Our results showed that atrazine exposure had a profound negative influence on selected indices and also on histological changes of common carp.

Effects of subchronic exposure of diclofenac on growth, histopathological changes, and oxidative stress in zebrafish (Danio rerio).

The aim of this study was to investigate effects of subchronic exposure to sublethal levels of diclofenac on growth, oxidative stress, and histopathological changes in Danio rerio. The juvenile growth tests were performed on Danio rerio according to OECD method number 215. Fish at the age of 20 days were exposed to the diclofenac environmental concentration commonly detected in the Czech rivers (0.02 mg L(-1)) and the range of sublethal concentrations of diclofenac (5, 15, 30, and 60 mg L(-1)) for 28 days. A significant decrease (P < 0.01) in the fish growth caused by diclofenac was observed in the concentrations of 30 and 60 mg L(-1). The identified value of LOEC (lowest observed effect concentration) was 15 mg L(-1) of diclofenac and NOEC (no observed effect concentration) value was 5 mg L(-1) of diclofenac. We did not find histopathological changes and changes of selected parameters of oxidative stress (glutathione S-transferase, glutathione reductase) in tested fish. The environmental concentration of diclofenac in Czech rivers did not have any effect on growth, selected oxidative stress parameters (glutathione S-transferase, glutathione reductase), or histopathological changes in Danio rerio but it could have an influence on lipid peroxidation.