Enhanced growth and decreased mortality in Atlantic salmon (Salmo salar) after probiotic bath.

AIMS: Disease in farmed Atlantic salmon occurs in all its life stages. Salmon are particularly vulnerable to infectious diseases at transition from the freshwater stage to the saltwater stage. Our aim in these studies reported was to investigate the possibility that waterborne delivery of a probiotic comprised of naturally occurring marine bacterial species would reduce the mortality and improve the health and growth of farmed Atlantic salmon. METHODS AND RESULTS: In three trials at two aquaculture production sites in Norway, isolates of Aliivibrio bacteria were added to the rearing water of Atlantic salmon. The fish were followed in 4-6 months after one single bath with observations and samplings. Growth, ulcers and survival were recorded. At the end of the studies growth was up to 31% larger in the probiotic enhanced groups and in trial 1 both mortality and prevalence of ulcer were significantly lower in the probiotic enhanced group compared to the control. Feed conversion rates were recorded in trial 1 and 2 and were from 9 to 28 % better for the probiotic enhanced groups compared to the control groups. CONCLUSION: Bathing of Atlantic salmon with probiotic Aliivibrio strains increased growth, reduced mortality and improved FCR in the postsmolt period. SIGNIFICANCE AND IMPACT OF THE STUDY: The study demonstrates the potential to enhance growth, prevent ulcers and decrease mortality in Atlantic salmon after adding probiotic strains of Aliivibrio spp. into the rearing water. The study can have impact on animal welfare, economy and sustainability in the aquaculture industry.

The effect of a two-year school-based daily physical activity intervention on a clustered CVD risk factor score-The Sogndal school-intervention study.

To evaluate changes in clustered cardiovascular disease (CVD) risk factors in 9-year-old children following a 2-year school-based physical activity intervention. In total, 259 children (age 9.3 ± 0.3 years) were invited, of whom 256 participated. The intervention group (63 boys, 62 girls) carried out 60-minute teacher-controlled daily physical activity over two school years. The control group (62 boys, 69 girls) had the curriculum-defined amount of physical education (45 minutes twice each week). Of these, 67% (171 total, 91 intervention) successfully completed both baseline and post-intervention of six CVD risk factors: systolic blood pressure (SBP), triglyceride (TG), total cholesterol-to-high-density lipoprotein cholesterol ratio (TC:HDL ratio), waist circumference (WC), the homeostasis model assessment for insulin resistance (HOMA), and peak oxygen uptake (VO2peak ). All variables were standardized by sex prior to constructing a cluster score (sum of z scores for all variables). The effect of the intervention on the cluster score was analyzed using linear multiple regression. The cluster score improved after the intervention (ES = .29). Furthermore, the analyses showed significant effects in favor of the intervention group for systolic blood pressure (ES = .35), total cholesterol-to-HDL-c ratio (ES = .23), triglyceride (ES = .40), and VO2peak (ES = .57). A teacher-led school-based physical activity intervention that is sufficiently long and includes a substantial amount of daily physical activity can beneficially modify children's clustered CVD risk profile.

Objectively measured sedentary time and physical activity and associations with body weight gain: does body weight determine a decline in moderate and vigorous intensity physical activity?

OBJECTIVES: High levels of physical activity (PA) may prevent the development of obesity. However, the magnitude and direction of the association between PA of various intensities, sedentary time and weight status remain unclear. Thus, we examined whether objectively measured sedentary time and PA independently predict gain in body weight, change in body weight and to examine the possibility of reverse causation. METHODS: We examined the prospective associations between sedentary time, PA and body weight (BW). Baseline measurements were conducted in 2008/2009 and follow-up measurements in 2014/2015 in a random sample of the adult Norwegian population (N=1710, 45.1% men). Moderate and vigorous intensity PA (MVPA) and sedentary time were measured by accelerometry and BW and height self-reported. We first modelled the associations between baseline sedentary time and PA with BW at follow-up. We then modelled the reverse associations (BW as the exposure) and sedentary time and PA (as outcomes) in separate models. All models were adjusted for age, sex, baseline value of the outcome, socio-economic status, alcohol consumption, smoking, monitor wear time and follow-up time. RESULTS: Body mass index (BMI) increased by 0.2 units (P=0.003) between baseline and follow-up, and 46.5% of participants were either overweight (36.4%) or obese (10.1%) at baseline increasing to 49.6% (11.7% obese) at follow-up. Baseline sedentary time, MVPA and vigorous PA were not associated with BW at follow-up after adjustment for covariates. In contrast, baseline BW was inversely associated with MVPA (ß=-0.11; 95% confidence interval (CI); -0.21, -0.009) and VPA (ß=-0.035; 95% CI; -0.059, -0.011) in adjusted models. These associations were unchanged when BW was substituted by BMI. CONCLUSIONS: Baseline BW seems to determine a decrease in MVPA in healthy adult Norwegian men and women, more so than the reverse.

Aeromonas salmonicida infection levels in pre- and post-stocked cleaner fish assessed by culture and an amended qPCR assay.

Due to increasing resistance to chemical therapeutants, the use of 'cleaner fish' (primarily wrasse, Labridae, species) has become popular in European salmon farming for biocontrol of the salmon louse, Lepeophtheirus salmonis (Krøyer). While being efficient de-licers, cleaner fish mortality levels in salmon cages are commonly high, and systemic bacterial infections constitute a major problem. Atypical furunculosis, caused by Aeromonas salmonicida A-layer types V and VI, is among the most common diagnoses reached in clinical investigations. A previously described real-time PCR (qPCR), targeting the A. salmonicida A-layer gene (vapA), was modified and validated for specific and sensitive detection of all presently recognized A-layer types of this bacterium. Before stocking and during episodes of increased mortality in salmon cages, cleaner fish (primarily wild-caught wrasse) were sampled and screened for A. salmonicida by qPCR and culture. Culture indicated that systemic bacterial infections are mainly contracted after salmon farm stocking, and qPCR revealed A. salmonicida prevalences of approximately 4% and 68% in pre- and post-stocked cleaner fish, respectively. This underpins A. salmonicida's relevance as a contributing factor to cleaner fish mortality and emphasizes the need for implementation of preventive measures (e.g. vaccination) if current levels of cleaner fish use are to be continued or expanded.

Still childless at the age of 28 to 40 years: a cross-sectional study of Swedish women's and men's reproductive intentions.

BACKGROUND: Delayed childbearing is associated with adverse reproductive outcomes. Our aim was to investigate Swedish women's and men's childbearing intentions at the age of 28, 32, 36 and 40 years, in terms of: (1) time point for a first child, (2) number of children, and (3) reasons for not yet having children. METHODS: Cross-sectional data from the Swedish Young Adult Panel Study, including 365 childless women and 356 childless men aged 28, 32, 36 and 40 years who responded to a questionnaire in 2009. Descriptive and multivariate logistic regression analyses were conducted. RESULTS: Most 28- and 32-year-olds intended to have children, but only 32% of women and 37% of men aged 36/40 years (merged), many of whom still postponed childbearing. Reasons for remaining childless differed by age. Most prominent in the 36/40-year-olds were: lack of a partner (women 60%, men 59%), no desire for children (women 44%, men 44%), not mature enough (women 29%, men 35%), and wanting to do other things before starting a family (women 26%, men 33%). The 36/40-year-olds had the highest odds for infertility problems (OR 3.8; CI 95% 1.8-7.9) and lacking a suitable partner (OR 1.8 CI 95% 1.1-3.0), and lower odds for reasons related to work and financial situation. CONCLUSIONS: Many childless 36- and 40-year-olds intended to have children but seemed to overestimate their fecundity. The most prominent reasons for being childless were: not having wanted children up to now, lack of a partner, infertility problems, and prioritising an independent life.

Nitrogen dioxide: no influence on allergic sensitization in an intranasal mouse model with ovalbumin and diesel exhaust particles.

The role of traffic-related air pollution in the development of allergic diseases is still unclear. We therefore investigated if NO2, an important constituent of traffic-related air pollution, promotes allergic sensitization to the allergen ovalbumin (OVA). We also examined if NO2 influenced the allergy adjuvant activity of diesel exhaust particles (DEP). For this purpose, mice were exposed intranasally to OVA with or without DEP present, immediately followed by exposure to NO2 (5 or 25 parts per million [ppm]) or room air for 4 h in whole body exposure chambers. Eighteen hours after the last of three exposures, the lungs of half of the animals were lavaged with saline and markers of lung damage and lung inflammation in the bronchoalveolar lavage fluid (BALF) were measured. Three weeks later, after intranasal booster immunizations with OVA, the levels of OVA-specific IgE and IgG2a antibodies in serum were determined. Both NO2 (25 ppm) and DEP gave lung damage, measured as increased total protein concentration in BALF, whereas only NO2 seemed to stimulate release of the proinflammatory cytokine tumor necrosis factor alpha (TNF-α). In contrast, only DEP significantly increased the number of neutrophils. Furthermore, DEP in combination with OVA stimulated the production of serum allergen-specific IgE antibodies. NO2, however, neither increased the production of allergen-specific IgE antibodies, nor influenced the IgE adjuvant activity of DEP. Thus, based on our findings, NO2 seems to be of less importance than combustion particles in the development of allergic diseases after exposure to traffic-related air pollution.

European guideline for the management of genital herpes, 2010.

This is the guideline for genital herpes simplex virus (HSV) management for the IUSTI/WHO Europe, 2010. They describe the epidemiology, diagnosis, clinical features, treatment and prevention of genital HSV infection. They include details on the management of HSV in pregnancy, those who are immunocompromised and the clinical investigation and management of suspected HSV-resistant disease.

Beryllium sensitivity among workers at a Norwegian aluminum smelter.

BACKGROUND: Sensitivity to beryllium was investigated among workers at an aluminum smelter in Norway as a consequence of the findings in an occupational exposure survey. METHODS: Three hundred and sixty-two employees and 31 reference persons were tested for sensitization to beryllium with the beryllium lymphocyte proliferation test (BeLPT) based on specifications by the US Department of Energy in 2001. The results are reported as abnormal, borderline, or normal. RESULTS: One person (0.28%) from the aluminum smelter was found to have abnormal results in two separate blood samples and is sensitized to beryllium. Three other persons had one abnormal test that was not confirmed by a second test. One person in the reference group had one abnormal and one normal test result. No borderline samples were detected. None of the employees with one or more abnormal sample results had pot room asthma. The sensitized individual worked in a Soederberg line in 1972-1974. The beryllium concentration in the work atmosphere is estimated to have been similar as today (0.1-0.3 microg/m(3)), but work routines, etc. would cause higher total exposures. CONCLUSIONS: Only one sensitized person of 362 is in line with what is found in other studies in the aluminum industry. The low number, compared with the beryllium handling industry, may be attributable to lower work atmosphere concentrations, beryllium speciation effects, or use of respiratory protection equipment. Pot room asthma does not appear to be associated with beryllium sensitization.

Monocyte exposure to Saccharomyces cerevisiae cell wall mannan and decreased TNF-alpha production in mild asthma: a role for mannan-binding lectin?

A correlation between fungal exposure and aggravation of inflammatory symptoms in asthmatic individuals is well documented in the literature. However, the molecular mediators responsible for clinical symptoms due to fungal exposure in individuals with asthma are still not known. The fungal cell wall polysaccharide mannan stimulates production of pro-inflammatory cytokines such as tumour necrosis factor (TNF)-alpha in monocytes. Recently, a role for the plasma protein mannan-binding lectin (MBL) has been proposed in individuals with severe asthmatic disease, although little is known about its role in those with mild and untreated asthma. MBL has been reported to modulate inflammatory cytokine production, but the mechanisms are not known. We conducted a pilot study and found that the cell wall mannan preparation used stimulated lower TNF-alpha production by monocytes from asthmatic subjects compared with that from healthy subjects in the presence of autologous plasma. Lipopolysaccharide-induced TNF-alpha production was not significantly different between the groups. Further, plasma MBL levels in individuals with mild asthma were slightly increased compared with those in normal subjects, although the difference was not statistically significant. We speculate that reduced TNF-alpha production in monocytes from asthmatic subjects after fungal cell wall mannan stimulation could partly be influenced by plasma components such as MBL.

Front-face fluorescence measurement of photosensitizers and lipid oxidation products during the photooxidation of butter.

This paper shows that fluorescence spectroscopy can measure both degradation of photosensitizers and formation of lipid oxidation products in light-exposed butter. The photosensitizers were already notably degraded after 4 h of light exposure, whereas fluorescent lipid oxidation products were detected after 5 d. The fluorescence measurements were highly correlated with sensory assessments of acidic and rancid flavor. Photosensitizer degradation is therefore a promising indirect indicator of the onset of lipid oxidation in butter. Sensory analysis and measurement of peroxide value showed that the level of lipid oxidation was significantly higher for butter stored in air compared with butter stored in nitrogen (N2). This might be explained by the formation of singlet oxygen from direct photooxidation and type II photosensitized oxidation. Addition of the singlet oxygen quencher beta-carotene reduced the rancid flavor intensity in the air and N2 packages from 9.0 to 4.9 and from 6.5 to 4.7, respectively. Results indicate that lipid oxidation in the butter stored in N2 was mainly caused by type I photosensitized reactions, because addition of beta-carotene had little effect on the rancid flavor intensity.

Mycotoxin-induced depletion of intracellular glutathione and altered cytokine production in the human alveolar epithelial cell line A549.

Mould exposure has been associated with asthma and other inflammatory airway conditions. However, cellular effects of inhaled mould components are not well understood. We hypothesised that host defence mechanisms, such as production of cytokines (TGFbeta1, IL-6 and IL-8) and the intracellular antioxidant glutathione (GSH), could be adversely affected by different concentrations of mycotoxins. We studied the effects of citrinin and gliotoxin on lipopolysaccharide (LPS)-stimulated alveolar epithelial cells (A549). Cytokines in cell culture supernatants were analysed by ELISA and levels of GSH were measured by colorimetric (absorbance) determination. We found that GSH decreased in a dose- and time-dependent manner when cells were exposed to citrinin in particular. TGFbeta1 was moderately reduced at low mycotoxin concentrations but elevated at higher sub-toxic concentrations. A tendency for an inverse relationship between TGFbeta1 and GSH levels was observed. IL-6 and IL-8 were not significantly reduced at non-toxic mycotoxin concentrations. Thus, reduced epithelial GSH and TGFbeta1 levels combined with elevated IL-6 and IL-8 levels may result in increased pro-inflammatory activity during mycotoxin exposure. We suggest that this mechanism can contribute to inflammation in mould exposure.

Active photosensitizers in butter detected by fluorescence spectroscopy and multivariate curve resolution.

In this study, fluorescence excitation and emission matrices and multivariate curve resolution (PARAFAC) were used to detect and characterize active photosensitizers spectrally in butter. Butter samples were packed under high (air) and low oxygen (<0.05%) atmospheres and exposed to violet, green, or red light. Six photosensitizers were found: riboflavin, protoporphyrin, hematoporphyrin, a chlorophyll a-like molecule, and two unidentified tetrapyrrols. By estimation of relative concentrations, we could follow how each sensitizer was photodegraded as function of wavelength, oxygen level, and time. The degradation rate of protoporphyrin, hematoporphyrin, chlorophyll a, and one of the tetrapyrrols correlated well (0.83-0.91) with the formation of sensory measured oxidation. The results suggest that mainly type I photoreactions were responsible for the degradation of photosensitizers in both high and low oxygen atmosphere. Type II photoreactions (generation of singlet oxygen) were involved in the oxidation of butter stored in air. The study shows that PARAFAC modeling of fluorescence landscapes is an excellent tool for studying photooxidation in complex systems.

Measurement of lipid oxidation and porphyrins in high oxygen modified atmosphere and vacuum-packed minced turkey and pork meat by fluorescence spectra and images.

This paper illustrates that fluorescence spectroscopy and imaging can be used to measure the extent and distribution of lipid oxidation in meat. Minced turkey thighs and pork semimembranosus muscles were stored for 7 and 12 days at 4°C in high oxygen (O(2)) modified atmosphere packages and vacuum. Turkey meat packed in high O(2) atmosphere was oxidised already after 7 days of storage. The sensory rancid odour score was 4.7 (on a scale from 1 to 9) and the TBARS value was 1.86mg MDA/kg. There was also an increase in fluorescence emission intensity in the 410-550nm region, which arises from lipid oxidation products. The combination of unsaturated fatty acids and access to O(2) resulted in lipid oxidation gradients in the turkey meat samples, and these gradients were clearly visualised by fluorescence images. In comparison, pork meat was more stable against lipid oxidation, with TBARS values <0.2mg MDA/kg and no development of fluorescent lipid oxidation products was detected. The fluorescence spectra measured in the present experiment suggest that turkey thighs and pork semimembranosus muscle in addition to protoporphyrin also have a natural content of Zn protoporphyrin. The porphyrin content was higher in pork meat than in turkey meat. It increased during storage time when the meat was packed in vacuum, and it decreased with O(2) availability. The distribution of porphyrins in the meat was visualised by fluorescence imaging.

Prevalence of, and risk factors for, HSV-2 antibodies in sexually transmitted disease patients, healthy pregnant females, blood donors and medical students in Tanzania and Norway.

The prevalence of specific HSV-2 antibodies was studied in Tanzanian and Norwegian sexually transmitted disease (STD) patients (1095) and non-STD patients (488). Correlates to demographic and behavioural factors were evaluated. Seropositivity was determined by the non-commercial peptide-55 enzyme-linked immunoassay. The prevalence of HSV-2 antibodies was 70% in Tanzanian and 17% in Norwegian STD patients, 35% in Tanzanian blood donors and pregnant women, and 4, 7 and 14% in Norwegian medical students, blood donors and pregnant women respectively. A higher HSV-2 prevalence was associated with female sex, increasing age, previous STDs, history of genital HSV infection, coitarchal age (age at first intercourse) <15 years and HIV seropositivity. Compared to previous data, the prevalence of HSV-2 antibodies in Tanzanian STD patients has increased remarkably. In Norwegian STD patients our results are consistent with, or lower than, the prevalence previously reported in Western Europe. Demographic rather than behavioural factors were associated with higher prevalence of HSV-2 antibodies in STD patients.

The mycotoxins citrinin and gliotoxin differentially affect production of the pro-inflammatory cytokines tumour necrosis factor-alpha and interleukin-6, and the anti-inflammatory cytokine interleukin-10.

BACKGROUND: Microbial growth is considered one of the major causes of indoor air problems. Moulds have been associated with asthma, allergy and a wide range of diffuse indoor air-related symptoms. However, mechanisms of the adverse health effects are not well understood. OBJECTIVE: We hypothesized that the mycotoxins citrinin and gliotoxin could cause an imbalance between the secretion of the pro-inflammatory cytokines TNF-alpha and IL-6 and the anti-inflammatory cytokine IL-10. METHODS: We investigated the influence of citrinin and gliotoxin on the human monocytic cell line Mono-Mac-6 (MM6) with and without lipopolysaccharide -stimulation. The levels of IL-10, IL-6 and TNF-alpha were analysed in cell culture supernatants by ELISA. Cell viability and cell apoptosis were measured by flow cytometry. RESULTS: The strongest inhibition of cytokine secretion was found for IL-10. IL-6 levels were found to decrease in a dose-dependent manner along with reduced cell viability. TNF-alpha levels increased with low gliotoxin exposure (less than 100 ng/mL), but decreased significantly at 375 ng/mL and higher along with increased cell apoptosis and reduced cell viability. TNF-alpha levels were not reduced by citrinin exposure. CONCLUSION: We observed a cytokine imbalance with a more pronounced reduction of IL-10 concentrations compared with those of TNF-alpha and IL-6. We suggest that low exposure doses of citrinin and gliotoxin (corresponding to less than 100 ng/mL gliotoxin and less than 10 mug/mL citrinin) may inhibit IL-10 and lead to increased risk of an inflammatory response with relative overproduction of TNF-alpha and IL-6. The findings and their clinical implications must be verified by human studies. However, we speculate that the observed biological effects may be of importance as they may partly explain the occurrence of diffuse general indoor air-related symptoms as well as the worsening of asthmatic inflammatory reactions experienced in mouldy environments.

Reduction of IL-12 p40 production in activated monocytes after exposure to diesel exhaust particles.

BACKGROUND: A reduction of IL-12 production by lung macrophages may partly explain the presumed adjuvant effect of diesel exhaust particles (DEP) in allergy and asthma. IL-12 stimulates T helper type 1 (Th1) lymphocytes, which inhibit Th2 cells via Th1-specific cytokines. The aim of this study was to investigate the influence of DEP on the production of IL-12 p40 in lipopolysaccharide (LPS)-activated monocytes. METHODS: The human monocytic cell line Mono-Mac-6 was stimulated with LPS (200 ng/ml) and grown with DEP (0-200 microg/ml) for 0, 6 or 24 h. IL-12 p40 and the pro-inflammatory cytokine TNF were analysed in the cell supernatants by ELISA and a cell assay, respectively. RESULTS: Levels of IL-12 p40 correlated inversely with the DEP exposure concentrations, whereas TNF increased in parallel to the DEP concentrations. At a DEP concentration of 200 microg/ml, the amount of IL-12 p40 was 35% of that observed without DEP. The corresponding TNF value was 230% of the control. Reduced viability, binding of cytokines to DEP or endotoxin in the DEP samples cannot fully explain the changes in the concentrations of these two cytokines. CONCLUSION: DEP seem to inhibit the production of IL-12 p40 and stimulate that of TNF in activated monocytes. This may partly explain the presumed adjuvant effect of DEP in atopy; by altering the Th1/Th2 balance via down-regulation of IL-12, the Th2 response characteristic of allergy and asthma may be favoured.

Changing trends in genital herpes simplex virus infection in Bergen, Norway.

AIM OF STUDY: To document the proportion of each herpes simplex virus (HSV) type in genital HSV infection and changes over time during a 10 year period. DESIGN: Retrospective comparative study in sexually transmitted disease (STD) patients with genital HSV infection at the outpatient clinic for STD, Haukeland Hospital, Bergen, Norway. RESULTS: HSV-2 was the major cause during the 80's, whereas HSV-1 constitutes a greater part of the cases during the 90's, especially in female patients and in the younger age groups with primary or initial disease, where HSV-1 is the causative viral type in up to 70-90% of the cases. CONCLUSION: The documented change from HSV-2 towards HSV-1 in cases of genital HSV infection may have implications as to prognosis, future usefulness of vaccines, present and future usefulness of new type-specific serological tests.

Investigating the effect of clustering of the urban field on sustainable population growth of centrally located and peripheral towns.

In a previous study (Portnov and Erell, 1998a), an Index of Clustering was defined, which allowed an analysis of the combined effect of spatial isolation and remoteness of peripheral towns on the long-term patterns of their population growth. In the present paper, the analysis of the effect of clustering of the urban field on the patterns of population growth is extended to centrally located urban places, and the validity of this index is tested in two unevenly populated countries--Israel and Norway. In both countries, the effect of clustering of the urban field on the patterns of urban growth is twofold. In sparsely populated areas, the presence of neighboring towns appears to increase their chances of attracting potential migrants due to inter-urban exchanges, while in more densely populated areas, increasing clustering tends to reduce migration influx to a given locality due to inter-town competition. Following this conclusion, a strategy of "redirecting priorities" leading to the formation of urban clusters is proposed, which may enhance the potential of urban growth in geographical areas where this is desirable.

Short-term exposure of rodents to diesel exhausts: usefulness for studies of genotoxic and immunotoxic effects.

An exposure facility was tested with regard to the information obtainable from short-term animal experiments for the assessment of health hazards from automotive engine exhausts. Indicators of immunotoxicity and genotoxicity were studied in guinea pigs and mice, respectively, exposed for 2 weeks, 8 h/day, to ten times diluted exhausts from a one-cylinder research diesel engine running at constant load. Regulated and non-regulated pollutants were determined. Besides increased number of lavageable cells in the airways, exposed guinea pigs exhibited, after immunization and challenge to ovalbumin, reduced leukotrienes B4 and C4 in lavage fluid and reduced anti-ovalbumin IgG in serum. Absence of increased CYP1A activity indicated that the exposure was below the threshold for induction of these enzymes. Instead a certain reduction of this activity indicated interaction with active enzyme sites. In vivo doses of some reactive metabolites of low molecular mass were measured by adducts to hemoglobin. Doses from aliphatic epoxides were low, in accordance with low hydrocarbon levels in the exhaust. The levels of hemoglobin adducts from aldehydes showed no clearcut influences of exposure. Genetic effects determined by DNA fingerprint analysis were indicated. It is concluded that repeated dose inhalation exposure of small numbers of animals is a useful mode of exposure for studying parameters that may elucidate toxic effects of air pollutants emitted from automotive engines, with a possibility to evaluate engine and fuel with regard to health hazards.

Measurement of HPRT mutations in splenic lymphocytes and haemoglobin adducts in erythrocytes of Lewis rats exposed to ethylene oxide.

Young adult male Lewis rats were exposed to ethylene oxide (EO) via single intraperitoneal (i.p.) injections (10-80 mg kg-1) or drinking water (4 weeks at concentrations of 2, 5, and 10 mM) or inhalation (50, 100 or 200 ppm for 4 weeks, 5 days week-1, 6 h day-1) to measure induction of HPRT mutations in lymphocytes from spleen by means of a cloning assay. N-ethyl-N-nitrosourea (ENU) and N-(2-hydroxyethyl)-N-nitrosourea (HOENU) were used as positive controls. Levels of N-(2-hydroxyethyl)valine (HOEtVal) adducts in haemoglobin (expressed in nmol g-1 globin) were measured to determine blood doses of EO (mmol kg-1 h, mM h). Blood doses were used as a common denominator for comparison of mutagenic effects of EO administered via the three routes. The mean HPRT mutant frequency (MF) of the historical control was 4.3 x 10(-6). Maximal mean MFs for ENU (100 mg kg-1) and HOENU (75 mg kg-1) were 243 x 10(-6) and 93 x 10(-6), respectively. In two independent experiments, EO injections led to a statistically significant dose-dependent induction of mutations, with a maximal increase in MF by 2.3-fold over the background. Administration of EO via drinking water gave statistically significant increases of MFs in two independent experiments. Effects were, at most, 2.5-fold above the concurrent control. Finally, inhalation exposure also caused a statistically significant maximal increase in MF by 1.4-fold over the background. Plotting of mutagenicity data (i.e., selected data pertaining to expression times where maximal mutagenic effects were found) for the three exposure routes against blood dose as common denominator indicated that, at equal blood doses, acute i.p. exposure led to higher observed MFs than drinking water treatment, which was more mutagenic than exposure via inhalation. In the injection experiments, there was evidence for a saturation of detoxification processes at the highest doses. This was not seen after subchronic administration of EO. The resulting HPRT mutagenicity data suggest that EO is a relatively weak mutagen in T-lymphocytes of rats following exposure(s) by i.p. injection, in drinking water or by inhalation.