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Journal Article 2016; 37(Suppl 1): 38-44 PubMed PMID: 28263529 Citation  Keywords:  Animals, Anti-Bacterial Agents:toxicity, Ecosystem, Soil Pollutants:toxicity, Toxicity Tests, Water Pollutants, Chemical:toxicity,.   

OBJECTIVES: The aim of this study was to assess the ecotoxicity of selected antibiotics (i.e. penicillin G, vancomycin and tetracycline) using ecotoxicological tests. Tests were conducted on organisms representing all trophic levels of the aquatic ecosystem, namely producers (green freshwater algae Pseudokirchneriella subcapitata), consumers (water fleas Daphnia magna) and decomposers (bacteria Vibrio fischeri). The effect of antibiotics on the representative of edaphon was measured by testing the inhibition of the reproduction of springtails Folsomia candida and earthworms Eisenia fetida.

DESIGN: Methodologically, the procedure was carried out in accordance with the following standards: OECD 201 (Fresh water algal growth inhibition test), OECD 202 (Inhibition of the mobility of Daphnia magna), ISO 11348-2 (Inhibitory effect of antibiotics on the light emission of Vibrio fischeri), OECD 232 (Inhibition of reproduction of Collembola Folsomia candida) and OECD 222 (Inhibition of reproduction of Eisenia fetida).

RESULTS: In aquatic organisms the highest level of toxicity was shown by tetracycline to algae (72hEC50 = 1.82 mg.l-1) and daphnia (48hEC50 = 8.16 mg.l-1). The least toxic for all test organisms was penicillin G. The results of the tests performed on the representative of edaphon, Folsomia candida, showed that its reproduction was most inhibited by penicillin G (28dEC50 = 328 mg.kg-1) and least by tetracycline (28dEC50 = 2560 mg.kg-1). Similar results were observed in Eisenia fetida (56dEC50 = 348 mg.kg-1 for penicillin G and 56dEC50 = 2735 mg.kg-1 for tetracycline.

CONCLUSION: The ecotoxicity of antibiotics differed significantly depending on the test organism and testing conditions....

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Journal Article 2016; 37(3): 245-250 PubMed PMID: 27618600 Citation  Keywords:  Acquired Immunodeficiency Syndrome:microbiology, Adult, Anti-Bacterial Agents:pharmacology, Bacteria, Colonic Neoplasms:microbiology, Colorectal Neoplasms:microbiology, Female, Gentamicins:pharmacology, HIV Infections:microbiology, HL-60 Cells, Humans, In.   

OBJECTIVES: Bacteria from the intestinal tract of Slovak and American HIV/AIDS patients and Slovak colon cancer patients were tested for the capacity to be internalized by cells of the HL-60 cell line as well as by normal human lymphocytes. They were anticipated to possess a specific characteristic, i.e. a vigorous ability to be internalized by HL-60 cells and human lymphocytes. This assumption was confirmed by gentamicin protection assay.

RESULTS: Internalization of bacteria from HIV/AIDS patients frequently resulted in partial (patients SKM1, SKM22) or complete lysis (patients SKK1-1, SKM12) of HL-60 cells. In comparison with intramucosal bacteria isolated from patients with colorectal cancer (TSG, 883, 660, 838, 536, MZRa), their capacity to internalize HL-60 cells was found to be 15-20 times higher (USP15/7, USP1/4, USP3/3, SK725/5). Partial lysis (patients USP15/7, USP3/3 and SKM22) and complete lysis (patients USP1/4, SKK1-1/1, SKM1/6, SKM12/5) were detected also after internalization of bacteria by normal human lymphocytes. Compared to the amount of intracellular bacteria isolated from patients with HIV/AIDS, the ability of bacteria from patients with colorectal cancer to internalize normal human lymphocytes was significantly lower (10-15 times), yet still higher than that of bacteria isolated from healthy people.

CONCLUSIONS: Our results present the ability of bacteria of colon cancer patients and HIV/AIDS patients to internalize HL-60 cells and normal human lymphocytes. The findings underline the potentially important function of bacteria in the induction of colorectal cancer and immunodeficiency. The particularly high detection ability of bacteria from HIV/AIDS patients to internalize normal human cells emphasizes their potentially important role in the process of AIDS....

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Journal Article 2015; 36(Suppl 1): 79-87 PubMed PMID: 26757128 Citation  Keywords:  Animals, Anti-Bacterial Agents:toxicity, Body Size:drug effects, Carps:embryology, Embryo, Nonmammalian:drug effects, Fluoroquinolones:toxicity, Glutathione Peroxidase:drug effects, Glutathione Reductase:drug effects, Glutathione Transferase:drug effects,.   

OBJECTIVES: The aim of this study was to assess the effect of the subchronic exposure of early stages of common carp (Cyprinus carpio L.) to norfloxacin using morphometric data and oxidative stress parameters.

METHODS: A subchronic toxicity test was performed on fertilized embryos of common carp according to the OECD Guidelines No. 210. Embryos were exposed to norfloxacin concentrations of 0.0001 (environmental), 0.1, 1.0, 5.0, and 10.0 mg.L(-1) for 34 days.

RESULTS: At the end of the test (day 34), significant (p<0.05) stimulation of development was observed in all experimental groups, in contrast to the control. Significantly greater (p<0.01) total body length was also observed in the group exposed to 10.0 mg.L(-1) of norfloxacin compared to the control. A significant increase in the activity of glutathione S-transferase in all carp exposed to norfloxacin concentrations of 0.1 and 1.0 mg.L(-1) (p<0.01), and 5.0 mg.L(-1) (p<0.05) compared to control group was revealed. The activity of glutathione peroxidase was significantly lower (p<0.01) in experimental carp exposed to a norfloxacin concentration of 10.0 mg.L(-1). In experimental carp exposed to a norfloxacin concentration of 0.0001 mg.L(-1), a significant increase (p<0.05) in glutathione reductase activity was found. Significant (p<0.01) decreases in the content of thiobarbituric acid reactive substances in the groups exposed to norfloxacin concentrations of 1.0, 5.0, and 10.0 mg.L(-1) were revealed.

CONCLUSION: From the results, we can conclude that norfloxacin has a negative impact on selected biochemical processes related to the production of reactive oxygen species in early-life stages of common carp....

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Journal Article 2015; 36(Suppl 1): 100-105 PubMed PMID: 26757126 Citation  Keywords:  Anti-Bacterial Agents:analysis, Chromatography, High Pressure Liquid, Czech Republic, Drug Residues, Rivers:chemistry, Solid Phase Extraction, Sulfamerazine:analysis, Sulfamethazine:analysis, Sulfamethoxazole:analysis, Sulfonamides:analysis, Water Polluta.   

OBJECTIVES: The aim of this study was to assess the contamination level of aquatic ecosystems of the Oslava and the Jihlava Rivers, and of the Nove Mlyny Water Reservoir, situated in the South Moravian Region (Czech Republic), by residues of selected veterinary pharmaceuticals. We isolated and determined 10 sulfonamide antibiotics in samples of surface water and bottom sediments using optimized analytical methods.

DESIGN: A representative number of sampling sites in the entire basin of selected waters were chosen. Samples were collected particularly near the larger cities in order to assess their possible impact to the aquatic ecosystems. Extraction, pre-concentration and purification of samples were performed using optimized methods of solid phase extraction and pressurized solvent extraction. Final identification and quantification were carried out by high-performance liquid chromatography coupled with diode array detector.

RESULTS: The concentration of sulfonamides in water samples were all under the limit of detection. Regarding sediment samples, sulfadimidine was found at most sampling sites; its highest values were recorded in the Jihlava River (up to 979.8 µg.kg(-1) dry matter). Other frequently detected sulfonamides were sulfamethoxazole and sulfamerazine. Most other sulfonamides were under the limit of detection or limit of quantification.

CONCLUSIONS: Monitoring of antibiotic residues in the environment, especially in the aquatic ecosystem, is a current topic due to the growing worldwide use in both human and veterinary medicine. According to obtained results, we document the pollution of selected rivers and water reservoir by particular sulfonamides which basically reflects their application in veterinary medicine....

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Journal Article 2015; 36(Suppl 1): 57-63 PubMed PMID: 26757114 Citation  Keywords:  Animals, Arsenic:pharmacology, Bacterial Infections:blood, Carcinogens:pharmacology, Cyanobacteria, Erythrocyte Indices:drug effects, Iron:blood, Leukocyte Count, Microcystins:pharmacology, Microcystis, Neutrophils:drug effects, Oncorhynchus mykiss:blood,.   

OBJECTIVES: Under environmental conditions, fish are simultaneously exposed to multiple stressors. This study provides new knowledge on the effects of controlled exposure to multiple stressors, namely cyanobacterial biomass and food contaminated with arsenic.

METHODS: Rainbow trout were divided into six groups of 25 fish and exposed to different contaminant combinations for 30 days: 1) control group, 2) cyanobacterial biomass, 3 & 4) two groups exposed to arsenic at concentrations of 5 mg.kg(-1) and 50 mg.kg(-1) fish feed, and 5 & 6) two groups exposed to cyanobacterial biomass and arsenic combined. We then evaluated pathological, haematological and immunological parameters at 10, 20 and 30 days after exposure.

RESULTS: Marked gross pathological findings were present in groups exposed to arsenic and arsenic/cyanobacteria after 30 days. A strong decrease in haemoglobin concentration was observed in all experimental groups receiving arsenic after 10 days exposure. Total leukocyte count increased markedly in fish exposed to cyanobacterial biomass, and to higher arsenic concentrations by the end of the experiment. Neutrophils decreased significantly at the end of exposure. Similarly, exposure to cyanobacteria and/or arsenic led to suppression of opsonised zymosan particle-induced neutrophil respiratory bursts.

CONCLUSIONS: Our results demonstrate that the effects of exposure to toxic cyanobacterial biomass and arsenic on fish are enhanced when the contaminants are combined. In particular, long-term exposure led to disturbances in the white blood-cell count. Modulation of phagocytosis, which is the first line of defence against invading pathogens, suggests that the combined action leads to a decreased ability to control infection....

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Journal Article 2015; 36(5): 458-464 PubMed PMID: 26707047 Citation  Keywords:  Adolescent, Autoantibodies:immunology, Candida albicans, Candidiasis:immunology, Carrier State:immunology, Child, Child, Preschool, Cross Reactions, Female, Ghrelin:immunology, Growth Disorders:immunology, Helicobacter Infections:immunology, Helicobacter .   

OBJECTIVES: Many of peptides synthesized in gastrointestinal tract (GI) and adipose tissues, regulate growth and food intake. The GI microflora is an antigenic source. Based on the molecular mimicry hypothesis, intestinal microbe-derived antigens may trigger the production of autoantibodies cross-reacting with some neuropeptides.

DESIGN: The aim of the study was to assess whether in idiopathic short stature (ISS) children with Candida albicans (C.albicans) colonisation and/or Helicobacter pylori (H.pylori) infection the autoantibodies (in positive levels) against selected neuropeptides [anti-NP Abs(+)]: ghrelin, leptin, orexin A, αMSH are more prevalent than in Controls.

SETTING: The study group comprised 64 children with ISS and 36 children with normal height (Controls). In each child, IgG antibodies against H.pylori, ghrelin, leptin, orexin A and αMSH were assessed in serum, while presence of C.albicans - in stool samples.

RESULTS: The higher prevalence of anti-NP Abs(+) in ISS children with C.albicans and/or H.pylori than in normal height children with the colonization in question (34.4% vs 21.1%, p<0.01) was found. The prevalence of anti-NP Abs(+) in groups of children without C.albicans and H.pylori were low, anti-NP Abs(+) were detected in 9.4% of ISS children only, while in Controls they were not found.

CONCLUSIONS: In short children with C.albicans and/or H.pylori the incidence of autoantibodies against selected neuropeptides is high. It probably is connected with molecular mimicry between antigens of these microbiota and the mentioned peptides. It is tempting to speculate that presence of cross-reacting autoantibodies against regulatory neuropeptides may results in worse growth velocity. However, further studies are necessary to elucidate this issue....

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Journal Article 2014; 35(Suppl 2): 99-104 PubMed PMID: 25638373 Citation  Keywords:  Animals, Antibodies:immunology, Bacterial Infections:prevention & control, Cell Line, Chickens, Egg Yolk:immunology, Epithelial Cells:immunology, Humans, Immunoglobulin Fab Fragments:immunology, Immunoglobulins:immunology, Inflammation:immunology, Lung:im.   

OBJECTIVES: As an alternative therapeutic approach for the prevention and treatment of bacterial infections with P. aeruginosa of cystic fibrosis (CF) patients, chicken yolk antibodies (IgY) could be used. The most significant advantage of IgY, in contrast to mammalian IgG, consists in the fact, that when bound to the antigen, they usually do not induce inflammatory reaction. In addition, the simplicity of egg production and the ease of IgY preparation makes this kind of antibody an excellent tool for passive immunization. Thus, the aim of our project was to study the effect of IgY and its Fab fragment on the potential induction of pro-inflammatory reactions in lung epithelial cells.

METHODS: Chicken IgY were prepared from pooled egg yolks. Fab fragmens of IgY were purified from the papain digest of IgY using DEAE-Sephacel ion exchange chromatography. Their purity was verified by SDS electrophoresis in polyacrylamide gel. Immortalized human cell lines, CuFi (CF patient) and NuLi (healthy subject), and A549 (human adenocarcinoma cells) were exposed to IgY, Fab, OVA, LPS (positive control), PBS (negative control), and human and goat IgG for 24 hours. The concentration of pro-inflammatory cytokines TNF-α, IL-1β, IL-6 and GM-CSF were determined in cell media using the BioPlex method, which enables the quantification of multiple analytes simultaneously in one sample.

RESULTS: Our results show that i) the Fab fragment induced levels of some proinflammatory cytokines, when compared to the PBS control, whereas ii) chicken IgYs did not induce any notable production of pro-inflammatory cytokines in contrast to intense effect of LPS on TNF-α and GM-CSF. In summary, our data show that levels of all cytokines are comparable with physiological values in human serum except of IL-1β, which concentration in cell medium was markedly elevated by Fab fragment.

CONCLUSIONS: The present data indicate that IgY are not inflammatory for lung cells and thus they are possibly applicable for prevention of airway bacterial infections.

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Journal Article 2014; 35(Suppl 2): 64-70 PubMed PMID: 25638368 Citation  Keywords:  Animals, Anti-Bacterial Agents:toxicity, Ciprofloxacin:toxicity, Oxidative Stress:drug effects, Toxicity Tests, Subchronic, Zebrafish,.   

OBJECTIVES: The aim of this study was to assess the impact of ciprofloxacin in Danio rerio on growth, on the development of histopathological changes in selected organs (gills, kidney, liver), and on the activity of some oxidative stress markers during a 28-day toxicity test.

METHODS: Juvenile growth tests were performed on D. rerio according to OECD guideline No. 215. Fish at the age of 30 days were exposed for 28 days to a range of concentrations of ciprofloxacin (0.7 μg x L(-1)--environmental concentration, 100, 650, 1100 and 3 000 μg x L(-1)).

RESULTS: There were no significant differences between the specific growth rates of fish from the test groups and from the control group. Histopathological examination revealed no pathological changes in organs of fish exposed to any ciprofloxacin concentration. The activity of glutathione S-transferase increased at the ciprofloxacin concentrations of 0.7 and 100 μg x L(-1) compared to the control. A significant decrease in glutathione reductase activity was obtained in fish exposed to ciprofloxacin at 1100 and 3000 μg x L(-1); a significant decrease in glutathione peroxidase activity was also found, but at all tested concentrations except for 100 μg x L(-1). A decrease in the concentration of thiobarbituric acid reactive substances was noted only at 100 μg x L(-1) compared to the control.

CONCLUSIONS: According to our results, all tested concentrations of ciprofloxacin, including the environmental concentration, had an influence on oxidative stress markers and detoxifying enzymes in exposed fish, but did not affect fish growth or cause the development of histopathological changes in the fish organism.

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