Cytogenetic analysis of nuclear abnormalities in the erythrocytes of gecko lizards (Phyllopezus periosus) collected in a semi-arid region of northeast Brazil: Possible effects of natural background radioactivity.

High natural-background radioactivity levels occur in the semi-arid region of the State of Rio Grande do Norte, northeastern Brazil. We have studied the lizard Phyllopezus periosus, an endemic species of the Brazilian caatinga with saxicolous habitat, as a bioindicator of environmental quality. Specimens were collected in three areas, an environmental protection area and two areas recognized as having high natural background radiation, one of these being a mining area. Level of metals and gamma radiation emitters present in the water sources potentially used by the lizards were measured. The biological endpoints assessed were micronuclei and nuclear abnormalities in blood samples. Significant differences in background radioactivity levels were found among the assessed areas. Statistically significant differences in micronuclei and nuclear abnormality frequencies were seen, among the study areas and a relationship between radioactivity level and genetic damage was observed.

Modified comet assay with Giemsa staining: a suitable method for assessing genotoxicity in rats / Ensaio cometa modificado com coloração de Giemsa: um método adequado para avaliar a genotoxicidade em ratos

The present study tested a comet assay that was modified for compatibility with Giemsa staining to assess the drug genotoxicity in the peripheral blood of rats. We analysed the peripheral blood of 16 female Wistar rats (N=8 rats/group) from a control group and from a group that was treated with an intraperitoneal injection of 50mg cyclophosphamide/kg. The comet assay was carried out with modifications of the blood volume and immersion time in the lysing solution and different combinations of electrophoresis conditions (running time, voltage and current), to Giemsa staining. The lysing time and electrophoresis conditions allowed for the expression of all classes of DNA damage during the electrophoresis run, and the comets were efficiently stained with Giemsa. The technique showed high reproducibility for the DNA classes. The results demonstrate that the modified comet assay with Giemsa staining can be standardized for routine laboratory procedures using a 20µL blood sample, 3h and 30min immersions in the lysing solution and electrophoresis runs with 23 to 25 V and 310 and 360mA of electrical current. The modified comet assay with Giemsa staining that was described in the present study was standardized to be applied in the laboratory routine.(AU)

O presente estudo testou um ensaio cometa modificado para a coloração de Giemsa para avaliar a genotoxicidade de fármacos no sangue periférico de ratos. Analisou-se o sangue periférico de 16 ratas Wistar (n=8 ratas/grupo) de um grupo controle e de um grupo que foi tratado com uma injeção intraperitoneal de 50mg/kg pv. de ciclofosfamida. O ensaio cometa foi realizado com modificações do volume sanguíneo e do tempo de imersão na solução de lise, bem como com diferentes combinações de condições de eletroforese (tempo de corrida, tensão e corrente), para coloração de Giemsa. O tempo de lise e as condições de eletroforese permitiram a expressão de todas as classes de danos no DNA durante a corrida de eletroforese, e os cometas foram eficientemente corados com Giemsa. A técnica mostrou alta reprodutibilidade para as classes de DNA. Os resultados demonstram que o ensaio cometa modificado com coloração de Giemsa foi padronizado para procedimentos laboratoriais de rotina usando-se uma amostra de sangue de 20µL, 3h30min de imersão na solução de lise e eletroforese com 23 a 25 V e 310 e 360mA. O ensaio cometa modificado com coloração de Giemsa descrito foi padronizado para ser aplicado na rotina laboratorial.(AU)

Genotoxicity assessment of polluted urban streams using a native fish Astyanax altiparanae.

Water quality has declined globally due to increased contamination of aquatic ecosystems. The use of fish genotoxicity biomarkers may improve and complement parameters for environmental risk assessment. The aim of this study was to assess the genotoxicity of samples collected from streams of the Jordão River, a tributary of the Paranaíba River, Brazil with different levels of metal contamination, utilizing a native fish species to determine the sensitivity and viability of implementing a useful, reliable technique for routine biomonitoring programs. Chemical analysis of water and sediments collected from different sites indicated that a gradient of contamination existed as evidenced by different concentrations of metals detected. After chronic exposure to contaminated samples, micronucleus (MN) frequencies in fish erythrocytes were measured and correlation with environmental parameters determined. Sites where the water concentrations of the metals aluminum (Al), iron (Fe), manganese (Mn), zinc (Zn) and copper (Cu) were high indicating a greater genotoxic potential of these elements. At the samples collected from the urban zone, a gradual increase was found for chromium (Cr), cadmium (Cd) and nickel (Ni) indicative of adverse impacts of discharge of urban effluents. Data demonstrated that Astyanax altiparanae, used in the test, exhibited a reliable sensitivity for detection of genotoxic consequences attributed to exposure to water samples collected near the discharge of industrial and domestic waste.

Antioxidant activities of Moringa oleifera leaf extract against pendimethalin-induced oxidative stress and genotoxicity in Nile tilapia, Oreochromis niloticus (L.).

To assess the ameliorative effects of Moringa oleifera (MO) leaf extract on haematological and biochemical changes, liver DNA damage and oxidative stress biomarkers in Nile tilapia (Oreochromis niloticus) exposed to a sublethal concentration (0.52 mg/l) of pendimethalin (PM). Tilapia fish were allocated into four equal groups in tri-replicates as follows: first group was the control group, second group was treated with MO (20 ml/30 l water), third group was exposed to 0.52 mg PM/l and fourth group was exposed to 0.52 mg PM/l and treated with MO leaf extract (20 ml/30 l water) for 28 days. At the end of this period, blood and liver tissue samples were collected and haematological and biochemical changes, hepatic DNA fragmentation and oxidative stress biomarkers were analysed. Pendimethalin caused significant reduction in haematological profile [White blood cells (WBCs) and red blood cells (RBCs) counts, haemoglobin (Hb) concentration and haematocrit (Ht) level]; meanwhile, serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), creatinine, uric acid, glucose, cortisol, cholesterol and lactate dehydrogenase (LDH) were significantly increased. On the other hand, serum total protein, albumin, globulin and acetylcholinesterase (AChE) were decreased. Significant reduction in hepatic superoxide dismutase (SOD), catalase (CAT), total antioxidant capacity (TAC) and glutathione peroxidase (GSH-Px) levels and marked increments of hepatic malondialdehyde (MDA) and DNA fragmentation were observed in PM-exposed fish compared to the control group. The addition of Moringa oleifera leaf extract into the water could overcome the negative impacts of pendimethalin and normalise the examined parameters nearly to the control values. Moringa oleifera was used for the first time to protect tilapia fish against PM-induced toxicity. The present study revealed that Moringa oleifera has potent antioxidant and antigenotoxic actions against pendimethalin toxicity.

Beneficial effects of water-soluble chestnut (Castanea sativa Mill.) tannin extract on chicken small intestinal epithelial cell culture.

Feed and water supplementation with powdered hydrolyzable tannins from chestnut represents a valuable alternative strategy to antibiotics in animal nutrition. In this study, we evaluated the effects and safety of a water-soluble form of chestnut tannin (WST) in an in vitro model of chicken small intestinal epithelial cells (CSIEC). A chicken cell culture was established, and WST in concentrations of 0.025, 0.05, 0.1, and 0.2% were tested for cytotoxicity, cell proliferation, metabolic activity, production of reactive oxygen species, intracellular antioxidative potential, genotoxicity, and influence on the epithelia cell cycle. The tested concentrations showed a significant (P < 0.05) greater proliferative effect on CSIEC than the control medium (maximal proliferation at 0.1% WST as determined by optical density measurements). The 0.2% concentration of WST was cytotoxic, causing significantly higher (P < 0.05) nitric oxide and hydrogen peroxide production but with no short-term genotoxicity. Although increasing the concentration caused a decline in the metabolism of challenged cells (the lowest at 0.1% WST), metabolic activity remained higher than that in control cells. The antioxidant potential was 75% better and significantly (P < 0.05) higher in the 0.1% WST cultured cells compared to control. In conclusion, the cultured CSIEC are useful tools in basic and clinical research for the study of intestinal physiology, as they retain physiological and biochemical properties and epithelial morphology close to the original tissue and, in many ways, reflect the in vivo state. Our results indicate that WST exert a beneficial effect on intestinal epithelia, since they: i) stimulate proliferation of enterocytes; ii) increase antioxidative potential; iii) have no genotoxic effect; and iv) do not affect cellular metabolism. Our results reinforce the importance of WST as promising candidates for further evaluation and use in commercial broiler farm production.

Impact of acid mine drainage on haematological, histopathological and genotoxic effects in golden mahaseer, Tor putitora.

The present study was carried out to evaluate sub-lethal mechanism of acid mine drainage toxicity in fingerlings (9.5 ± 2.4 cm) of golden mahseer, Tor putitora. Exposed fingerlings showed significant reduction (P < 0.01) in blood erythrocytes, neutrophils, thrombocytes, lymphocytes and leukocytes in contrast to increase in number of immature circulating cells. Hyperplasia, degeneration of glomeruli, presence of inflammatory cells and increased number of melanomacrophage aggregates, vacuolization of cell cytoplasm, hepatocyte swelling were marked in kidney and liver of fish. Ladder in, an increment of 180-200 bp of hepatic and kidney DNA, by electrophoresis were consistent with DNA damage. 10 day exposure to acid mine drainage resulted in reduction of double stranded DNA to 46.0 and 48.0 in hepatocytes and kidney cells respectively. Significant increase (P < 0.01) in tail length and percent tail DNA was evident by comet assay. The results suggest that exposure to acid mine drainage might cause irreversible damage to immune cells, tissue and DNA of fish, and this model of DNA damage may contribute in identifying novel molecular mechanism of interest for bioremediation application.

Assessment of status of three water bodies in Serbia based on tissue metal and metalloid concentration (ICP-OES) and genotoxicity (comet assay).

Metals and metalloids are natural components of the biosphere, which are not produced per se by human beings, but whose form and distribution can be affected by human activities. Like all substances, they are a contaminant if present in excess compared to background levels and/or in a form that would not normally occur in the environment. Samples of liver, gills, gonads and muscle from European chub, Squalius cephalus, were analyzed for Al, As, B, Ba, Cr, Cu, Fe, Hg, Mn, Mo, Sr and Zn using inductively coupled plasma optical emission spectrometry (ICP-OES) to highlight the importance of tissue selection in monitoring research. The comet assay or single cell gel electrophoresis (SCGE) was selected as an in vivo genotoxicity assay, a rapid and sensitive method for measuring genotoxic effects in blood, liver and gills of the European chub. Microscopic images of comets were scored using Comet IV Computer Software (Perceptive Instruments, UK). The objective of our study was to investigate two reservoirs, Zlatar and Garasi, and one river, Pestan by: (i) determining and comparing metal and metalloid concentrations in sediment, water and tissues of European chub: liver, gills, muscle and gonads (ii) comparing these findings with genotoxicity of water expressed through DNA damage of fish tissues. A clear link between the level of metals in water, sediment and tissues and between metal and genotoxicity levels at examined sites was not found. This suggests that other xenobiotics (possibly the organic compounds), contribute to DNA damage.

Assessing genotoxic effects in fish from a marine protected area influenced by former mining activities and other stressors.

The goal of the current study was to evaluate different genotoxicity tools in order to assess a marine protected area (MPA) affected by former mining activities and urban settlements. A catfish (Cathorops spixii) was analyzed for genotoxic effects at the (i) molecular and at the (ii) chromosomal levels. Through factor analysis, genotoxicity was found to be linked to levels of metals bioaccumulated and PAH metabolites in the bile. Micronucleus and nuclear alteration were less vulnerable to the effects of confounding factors in mildly contaminated areas since they were more frequently associated with bioaccumulated metals than the DNA analysis. The different genotoxicity responses allowed for the identification of sources of pollution in the MPA. This approach was important for detecting environmental risks related to genotoxic contaminants in a mildly contaminated MPA.

Probiotic preparation reduces the faecal water genotoxicity in chickens fed with aflatoxin B1 contaminated fodder.

The aim of the study was to evaluate the influence of a probiotic preparation on the genotoxicity of faecal water of broiler chickens fed with a fodder contaminated with aflatoxin B(1) (AFB(1)) at 1 or 5mg per kg. Human blood lymphocytes were exposed to chicken's faecal water samples and DNA damage was measured using the comet assay. Genotoxicity of faecal water did not depend on the AFB(1) concentration in the fodder. The mean DNA damage, measured as the percentage of DNA in the tail of the comets, for chickens fed with fodder with AFB(1) at 1 mg/kg was 16.80±0.66, at 5 mg/kg - 16.73±1.51 and in the controls - 12.79±0.66. The supplementation of fodder with the probiotic preparation decreased the extent of DNA damage to 10.02±0.39 for 1 mg/kg AFB(1) and to 11.89±0.72 for 5 mg/kg.

An Assessment of Mutagenic Effect of 3, 3', 4, 4', 5 Pentachlorobiphenyl (PCB126) in Muta Mouse Fetuses.

PCBs are persistent environmental agents that induce multiple impairments in living beings. In this study we used a transgenic mouse model (Muta(TM) Mouse), carrying bacterial lacZ genes for mutation assays and for assessment of the genotoxic effect of PCB126 on fetal mice. Mothers of experimental groups were subjected to a single oral dose of PCB126 (125, 250 and 500 microg/kg) on the 10th day of pregnancy, respectively. Fetuses were autopsied on the 18th day of gestation. Cleft palate was observed in 2 out of 11 fetuses from 3 litters in 500 microg/kg treated group. Other external malformations were not observed. The DNA mutation frequencies (MF) of fetuses in each group were 1.15 +/- 0.24 x 10(-5), 0.90 +/- 0.20 x 10(-5) and 1.08 +/- 0.24 x 10(-5) in fetuses of 125, 250 and 500 microg/kg treated groups, respectively. The MF of controls was 0.81 +/- 0.22 x 10(-5). There were no significant differences among the groups. However, the MF of each treated group was a little highter than that of control group. Possible relationships between PCB and its mutagenic effects in the offspring of mice are discussed.

Single-dose pharmacokinetics and genotoxicity of metronidazole in cats.

Single-dose pharmacokinetics and genotoxicity of metronidazole in cats were evaluated. Cats received either 5mg/kg metronidazole intravenously, or 20mg/kg metronidazole benzoate (12.4mg/kg metronidazole base) orally in a single dose. Serial plasma samples were collected and assayed for metronidazole using high pressure liquid chromatography (HPLC). Genotoxicity was assessed in vitro in feline peripheral blood mononuclear cells (PBMC) and a feline T-cell lymphoma line incubated with metronidazole, and in vivo in PBMC collected before, during and 7 days after oral metronidazole, by use of the COMET assay. Systemic absorption of metronidazole was variable (mean=65+/-28%) with a peak of 8.84+/-5.4microg/ml at 3.6+/-2.9h. The terminal half-life was 5.34h from the intravenous dose and 5.16h from the oral dose. Systemic clearance was low (mean=91.57ml/h/kg [1.53ml/kg/min]), and the apparent volume of distribution (steady state) was 0.650+/-0.254l/kg. Genotoxicity was detected at all concentrations of metronidazole in feline PBMC and the T-cell lymphoma line in vitro. Genotoxicity was also observed in PBMC collected from cats after 7 days of oral metronidazole but resolved within 6 days of discontinuing metronidazole.

Evaluación genotóxica del D-004, extracto del fruto de Roystonea regia, mediante el ensayo de la morfología de la cabeza del espermatozoide en ratas Sprague Dawley / Genotoxic evaluation of D-004 in the Sprague Dawley rats sperm morphology assay

El tratamiento oral con D-004, extracto lipídico del fruto de la palma real (Roystonea regia), ha mostrado prevenir la hiperplasia prostática (HP) inducida por testosterona y por fenilefrina en ratas, efectos relacionados con su capacidad de inhibir la actividad de la 5 -reductasa prostática y de antagonizar los adreno-receptores 1.Este estudio, como parte de los estudios de genotoxicidad del D-004,tuvo como objetivo determinar si el mismo aumenta la frecuencia de aparición de formas anómalas de la cabeza del espermatozoide en ratas. Para ello se utilizaron ratas Sprague Dawley machos incluidas en un estudio de toxicidad crónica, distribuidas aleatoriamente en 5grupos (8 ratas/grupo): 1 control negativo tratado con el vehículo, 3grupos tratados con D-004 (800, 1500 y 2000 mg/kg, respectivamente), y 1 control positivo (Ciclofosfamida, 50mg/kg).Los tratamientos (vehículo o D-004) se administraron por vía oral durante 12 meses, la Ciclofosfamida se administró por vía intraperitoneal 5 días consecutivos. No ocurrieron muertes ni se detectaron signos de toxicidad. Las concentraciones de espermatozoides, y las frecuencias de espermatozoides normales en los grupos tratados con D-004 fueron similares a los del control negativo, mientras que el grupo control positivo mostró menor concentración de espermatozoides y mayor frecuencia de espermatozoides anómalos que los controles negativos. En conclusión, el D-004 administrado durante varios períodos de espermatogénesis no presenta potencial genotóxico sobre las células germinales masculinas de ratas (AU)

Oral treatment with D-004, a lipid extract from the royal palm(Roystonea regia) fruits, has been shown to prevent prostate hyperplasia (PH) induced by testosterone and phenylephrine in rats through the D-004-mediated inhibition of prostate 5 –reductase activity and antagonism of -1 adrenoceptors. This study, as part of the studies of the genotoxic potential of D-004, was aimed to determine whether D-004 increases the frequency of abnormal forms of rat sperm-head morphology. For that, male Sprague Dawley rats included in a chronic toxicity study were used, which were randomly distributed into 5 groups (8 rats/group): 1 negative control treatedwith the vehicle, 3 D-004-treated groups (800, 1500 and 2000 mg/kg, respectively), and 1 positive control (Ciclophosphamide, 50mg/kg).Treatments (vehicle or D-004) were orally administered for 12months, while Ciclophosphamide was given by intraperitoneal route five days consecutive. There were no deaths and no toxicity signs were observed. Spermatozoid concentrations and the frequencies of normal spermatozoids in D-004 treated groups were similar to those of the negative controls, while the positive control had decreased spermatozoid concentrations and increased frequency of abnormals permatozoids compared with the negative controls. In conclusion,D-004 orally administered during several periods of spermatogenesis in rats did not show genotoxic potential on the male germinal cells (AU)

Effects of cadmium on aneuploidy and hemocyte parameters in the Pacific oyster, Crassostrea gigas.

Pacific oysters, Crassostrea gigas, are commonly reared in estuaries where they are exposed to anthropogenic pollution. Much research has been made on the toxicity of cadmium to aquatic organisms because the compound recurrently contaminates their environment. Our study examined the influence of cadmium on aneuploidy level (lowered chromosome number in a percentage of somatic cells) and hemocyte parameters in C. gigas at different stages of life. Adults and juveniles were exposed to two different concentrations of cadmium. The first concentration applied was equivalent to a peak value found in Marennes-Oléron bay (Charente-Maritime, France; 50 ngL(-1)) and the second was 10 times higher (500 ngL(-1)). Exposure to 50 ngL(-1) cadmium caused a significant decrease in the survival time of C. gigas, but exposure to 500 ngL(-1) surprisingly affected the survival time positively. Significant differences in aneuploidy level were observed between the cadmium treatments and the control in adults but not in juveniles or the offspring of the adult groups. The effects of cadmium on hemocyte parameters were analyzed by flow cytometry. Several hemocyte parameters increased significantly after 21 days of cadmium exposure and subsequently decreased. Phenoloxidase-like activity, evaluated by spectrophotometry, varied over the time of the experiment and increased after 66 days of contact with 500 ngL(-1) cadmium. Taken together, cadmium at environmentally relevant concentrations seems to have only moderate effects on aneuploidy and hemocyte parameters.

Genotoxic and stress inductive potential of cadmium in Xenopus laevis larvae.

The present investigation evaluates the toxic potential of Cd in larvae of the frog Xenopus laevis after 12 days of exposure to environmentally relevant contamination levels, close to those measured in the river Lot (France). Several genotoxic and detoxification mechanisms were analyzed in the larvae: clastogenic and/or aneugenic effects in the circulating blood by micronucleus (MN) induction, metallothionein (MT) production in whole larvae, gene analyses and Cd content in the liver and also in the whole larvae. The results show: (i) micronucleus induction at environmental levels of Cd contamination (2, 10, 30 microgL(-1)); (ii) an increased and concentration-dependent quantity of MT in the whole organism after contamination with 10 and 30 microgCdL(-1) (a three- and six-fold increase, respectively) although no significant difference was observed after contamination with 2 microgCdL(-1); (iii) Cd uptake by the whole organism and by the liver as a response to Cd exposure conditions; (4) up-regulation of the genes involved in detoxification processes and response to oxidative stress, while genes involved in DNA repair and apoptosis were repressed. The results confirm the relevance of the amphibian model and highlight the complementarity between a marker of genotoxicity, MT production, bioaccumulation and genetic analysis in the evaluation of the ecotoxicological impact.

Effect of polychlorinated biphenyls (Delor 103) on haematological and enzyme parameters of the rainbow trout Oncorhynchus mykiss.

Rainbow trout at a weight of 223+/-12 g (mean+/-SD) were experimentally injected with a technical mixture of Delor 103 to evaluate the red blood cell indices (red blood cell count, haematocrit, haemoglobin, mean corpuscular volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration) and some biochemical and enzyme parameters of the blood plasma (total protein, glucose, inorganic phosphate, total calcium, sodium, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, lactate dehydrogenase). Delor 103, administered by the i.p. route at a concentration of 0.24 g kg(-1) 120 h(-1), caused an increase in the red blood cell counts, haematocrit values, haemoglobin concentrations, inorganic phosphate, alanine aminotransferase and lactate dehydrogenase. The sodium level fell. The fish injected with Delor 103 showed a relative decrease in the lymphocyte count and a relative increase in the count of neutrophile band forms.

Genotoxic, cytotoxic and ontogenetic effects of tri-n-butyltin on the marine worm, Platynereis dumerilii (Polychaeta: Nereidae).

The genotoxic, cytotoxic and ontogenetic (embryo-larval) or developmental effects of tri-n-butyltin (TBT), were investigated in Platynereis dumerilii. Following the determination of maximum tolerated dose with regard to ontogenetic effects and mortality, early life stages of P. dumerilii were exposed to a range of TBT concentrations. Subsequently, the embryo-larvae were analysed for evidence of genotoxicity and cytotoxicity. Genotoxicity was assessed using cytogenetic endpoints that included the frequency of sister chromatid exchanges and chromosomal aberrations from metaphase spreads. Cytotoxicity was evaluated by determining the proliferative rate index of the growing embryo-larval cells using 5-bromodeoxyuridine labelling of the chromosomes or fluorescence plus Giemsa staining technique. TBT-exposed embryo-larvae of P. dumerilii exhibited sensitivity similar to that of other invertebrates, indicating that P. dumerilii is a suitable ecotoxicity test species. The results also suggested dose-dependent effects for genotoxic and cytotoxic end points in relation to TBT exposure. The present study highlights the need to elucidate the relative importance of direct genotoxic and indirect effects through production of genotoxic hormonal derivatives.

The alterations in genetic apparatus of somatic and generative cells of the host caused by helminths metabolites.

Alterations in the genetic apparatus of mice bone marrow cells and testicles caused by Hymenolepis nana, Ascaris suum and Toxocara canis metabolites have been investigated by means of micronuclear test application. The activity of spermatogenesis has been investigated at experimental hymenolepidosis, migrating ascariasis and toxocarosis with the use of 3H-timidine. Helminths metabolites have been established to exert a mutagenic effect on somatic cells of bone marrow, spermatogonies and also on the generative cells (spermatides) of helminths in invaded mice. The concurrent increase in micronucleus number in erythrocytes, spermatogonies and in spermatides (to a lesser degree) of invaded mice has been revealed. The decrease in spermatogenesis activity has been established in experimental hymenolepidosis, migrating ascariasis and toxocarosis in invaded mice.

Inhibitory effect of dairy products on the mutagenicities of chemicals and dietary mutagens.

The antimutagenic effects of uninoculated milk and milks cultured with Bifidobacterium or Lactobacillus strains towards the mutagenicity induced by two direct mutagens, 4-nitroquinoline N-oxide and 2-nitrofluorene, and three dietary indirect mutagens, aflatoxin B1, benzo(a)pyrene and quercetin, were investigated using the in vitro Salmonella typhimurium test. Each cultured milk sample and control milk had a significant antimutagenic effect, to an extent varying with the mutagen used. Uninoculated milk had a greater inhibitory effect than cultured milks towards dietary indirect mutagens.

Mutagenicity, creatine and nutrient contents of pan fried meat from various animal species.

The mutagenic activity in extracts of fried meat from 16 different animal species was studied in Salmonella typhimurium TA98. In each experiment, 1 meat sample together with a standard beef sample was fried, and the mutagenicity was expressed relative to the beef sample. All meat samples showed less mutagenic activity than beef. The contents of creatine, creatinine, water, protein, carbohydrate and fat in the meat samples were analyzed, but mutagenicity was not correlated with the concentration of any of these constituents. Beef meat treated with creatinase to remove creatine produced reduced mutagenic activity. Possibly a threshold concentration of creatine is necessary to give a high mutagenic response.

Effect of chloramphenicol on sister chromatid exchange in bovine fibroblasts.

The genotoxic potential of different chloramphenicol concentrations (5, 20, 40 and 60 micrograms ml-1) was investigated in bovine fibroblast primary lines by sister chromatid exchange assay. Chloramphenicol acted for long enough to ensure similar effects to persistent storage in the kidney. In this experiment 10 micrograms ml-1 of 5-bromodeoxyuridine was added for 60 hours for all doses of chloramphenicol and to the control. When the tissue culture cells were exposed to increasing doses, increased numbers of sister chromatid exchanges developed. Differences were significantly different to the control.