Aksel Bernhoft

Organochlorines in polar bears (Ursus maritimus) at Svalbard

A comprehensive survey on organochlorine (OC) contaminants in polar bears at Svalbard has been undertaken. Subcutaneous tissue, blood and milk have been sampled from anesthetized free-ranging bears of both sexes and different ages in the period from 1990 to 1994. A number of sexually mature females have been fitted with satellite transmitters which make it possible to follow their reproductive behaviour pattern. We report on contamination levels and pattern of PCB congeners, chlordanes, DDT-compounds, HCB and HCH-isomers in the various tissues. The relation of age, sex and reproductive status to OC contamination has been described, and the capacity of polar bears in metabolising OCs has been discussed from the isomer/metabolite composition of residues. Finally, the possible association between OC contamination and reproduction success has been assessed. The PCB levels present in polar bears at Svalbard are extremely high. Especially high levels of higher chlorinated PCBs are found, and they accumulate with age, particularly in males. With females, considerable amounts of OCs are transferred to the offspring via milk. However, more efficient OC transfer between subcutaneous depot lipid and circulatory lipids than from the circulatory system to milk is found particularly for the most lipophilic compounds. The OC pattern in suckling yearlings reflects the low transfer of the highest chlorinated PCBs into maternal milk. The levels of most other OCs, however, are higher in depot lipid of yearlings than in that of their mothers. The polar bears have high capacity to metabolise several OCs. This may protect them against toxic action of the contaminants. No relation between OC levels in females and their ability to get pregnant is found in this preliminary investigation on OC effects on the polar bears at Svalbard.

Mycotoxin production and cytotoxicity of Fusarium strains isolated from Norwegian cereals.

Thirty-four isolates of the eight most common Fusarium species isolated from Norwegian cereals; F. avenaceum, F. culmorum, F. equiseti, F. graminearum, F. poae, F. sporotrichioides, F. torulosum and F. tricinctum were studied for their cytotoxicity and ability to produce mycotoxins. The strains were cultivated on rice, and analysed for trichothecenes (all species), zearalenone (all species), fusarochromanone (F. equiseti), wortmannin (F. torulosum), moniliformin and enniatins (F. avenaceum, F. tricinctum and F. torulosum). The cytotoxicity of the extracts were examined with an (in vitro) MTT-cell culture assay. All F. graminearum and five of seven F. culmorum isolates belonged to chemotype IA, producing deoxynivalenol and 3-acetyl-deoxynivalenol, while the two other F. culmorum strains were nivalenol producers (chemotype II). The F. equiseti isolates and one of the F. poae isolates produced both type A and B trichothecenes, and relatively large quantities of fusarochromanone were detected in the F. equiseti cultures. All Fusarium species studied showed significant cytotoxicity, but with a large variation between species, and also within each species. F. sporotrichioides and F. equiseti showed the highest average cytotoxicity.

Relationships between plasma levels of organochlorines, retinol and thyroid hormones from polar bears (Ursus maritimus) at Svalbard.

Associations were determined between retinol and the thyroid hormones thyroxine (T4) and triiodothyronine (T3), respectively, and the organochlorine contaminants (OCs) polychlorinated biphenyls (PCBs), 1,1-dichloro-2,2-bis-(4-chlorophenyl)ethylene (DDE), hexachlorobenzene (HCB), and hexachlorocyclohexanes (HCHs) in blood plasma from polar bears ( Ursus maritimus ) caught at Svalbard. The blood samples were collected from free-ranging polar bears of different age and sex in 1991-1994. The retinol concentration and the ratio of total T4 (TT4) to free T4(FT4) (TT4/FT4 ratio) decreased linearly with increasing concentrations of PCBs and HCB. Retinol was also negatively associated with HCHs, while the TT4/FT4 ratio was positively associated with DDE. The concentrations of retinol and thyroid hormones were significantly higher in females than in males. However, the TT4/FT4 and TT3/FT3 ratios were significantly higher in males than in females. The concentrations of thyroid hormones were negatively correlated with age in male bears, while in females, thyroid hormones did not change with age. The OCs were found to explain 12, 30, and 7% of the variation of retinol concentrations and the TT4/FT4 and TT3/FT3 ratios, respectively, after correcting for age and sex. The potential consequence of these associations for the individual and the population is unknown.

Possible immunotoxic effects of organochlorines in polar bears (Ursus maritimus) at Svalbard.

Associations between immunoglobulin G (IgG) levels and the organochlorine contaminants (OCs) polychlorinated biphenyls (PCBs), chlordanes, 1,1-dichloro-2,2-bis(4-chlorophenyl) ethylene (DDE), hexachlorobenzene (HCB), and hexachlorocyclohexanes (HCHs) in blood plasma from polar bears caught at Svalbard were determined. The blood samples were collected from free-living polar bears of different age and sex between 1991 and 1994. The IgC concentration increased with age and was significantly higher in males than in females. IgG was negatively correlated with sigmaPCB level and with three individual PCB congeners, IUPAC numbers 99, 194, and 206. HCB was also negatively correlated with IgG. The significant negative OC correlation with IgG levels may indicate an immunotoxic effect.

Organochlorines in top predators at Svalbard : occurrence, levels and effects

Alarmingly high polychlorinated biphenyl (PCB) levels have been found in the top predators such as glaucous gull (Larus hyperboreus) and polar bear (Ursus maritimus) at Svalbard [Gabrielsen, G.W., Skaare, J.U., Polder, A., Bakken, V., 1995. Chlorinated hydrocarbons in glaucous gull (Larus hyperboreus). Sci. Total Environ. 160/161, 337-346; Bernhoft, A., Skaare, J.U., Wiig, O., 1997. Organochlorines in polar bears (Ursus maritimus) at Svalbard. Environ. Pollut. 95, 159-175; Henriksen, E.O., Gabrielsen, G.W., Trudeau, S., Wolkers, H., Sagerup, K., Skaare, J.U., 1999. Organochlorines and possible biochemical effects in glaucous gull (Larus hyperboreus) from Bear Island, the Barents Sea. Arch. Environ. Contam. Toxicol. (in press). ]. Studies of the possible toxic effects, particularly on the immune system and reproduction, of the very high PCB levels in these species are currently being investigated. Data obtained in the field (f.i. reproductive success in polar bears and intestinal nematodes in glaucous gulls), as well as levels of various biochemical and physiological parameters (f.i. thyroid hormones, retinol, EROD activity, CYP1A, IgG), have been coupled with the PCB levels [Skaare, J.U., Wiig, O., Bernhoft, A., 1994. Klorerte organiske miljogifter; Nivâer og effekter i isbjorn. Norwegian Polar Institute Reportseries no. 86, 1-23 (in Norwegian); Bernhoft, A., Skaare, J.U., Wiig, O., 1997. Organochlorines in polar bears (Ursus maritimus) at Svalbard. Environ. Pollut. 95, 159-175; Bernhoft, A., Skaare, J.U., Wiig, O., Derocher, A.E., Larsen, H.J., 2000. Possible immunotoxic effects of organochlorines in polar bears (Ursus maritimus) at Svalbard (in press); Henriksen, E.O., Gabrielsen, G.W., Skaare, J.U., Skjegstad, N., Jensen, B.M., 1998a. Relationship between PCB levels, hepatic EROD activity and plasma retinol in glaucous gull, Larus hyperboreus. Marine Environ. Res. 46, 45-49; Henriksen, E.O., Gabrielsen, G.W., Trudeau, S., Wolkers, H., Sagerup, K., Skaare, J.U. , 1999. Organochlorines and possible biochemical effects in glaucous gull (Larus hyperboreus) from Bear Island, the Barents Sea. Arch. Environ. Contam. Toxicol. (in press); Sagerup, K., Gabrielsen, G.W., Skorping, A., Skaare, J.U., 1998. Association between PCB concentrations and intestinal nematodes in glaucou gulls, Larus hyperboreus, from Bear Island. Organohalogen compounds 39, 449-451; Skaare, J.U., Wiig, O., Bernhoft, A., 1994. Klorerte organiske miljogifter; Nivâer og effekter i isbjorn. Norwegian Polar Institute Reportseries no. 86, 1-23. (in Norwegian)].

Less Fusarium infestation and mycotoxin contamination in organic than in conventional cereals

A total of 602 samples of cereals, consisting of organically and conventionally produced barley, oats and wheat, were collected at harvest during 2002–2004 in Norway. Organic and conventional cereals were sampled in comparable numbers regarding cereal species, localisation and harvest time, and analysed for Fusarium mould and mycotoxins. Fusarium infestation and mycotoxin content were dependent on cereal species and varied year-by-year. However, in all cereal species, Fusarium infestation and levels of important mycotoxins were significantly lower when grown organically than conventionally. Concerning the most toxic trichothecenes, HT-2 and T-2 toxin, lower concentrations were found in organic oats and barley. Wheat was not contaminated by HT-2 and T-2, but lower concentrations of deoxynivalenol (DON) and moniliformin (MON) were found when organically produced. For mycotoxins considered to constitute the main risk to humans and animals in Norwegian cereals, i.e. HT-2 in oats and DON in oats and wheat, the median figures (mean levels in brackets) were as follows: HT-2 in organic and conventional oats were <20 (80) and 62 (117) µg/kg, DON in organic and conventional oats were 24 (114) and 36 (426) µg/kg, and DON in organic and conventional wheat were 29 (86) and 51 (170) µg/kg, respectively. Concentrations of HT-2 and T-2 in the samples were strongly correlated (r = 0.94). Other mycotoxins did not show a significant correlation to each other. Both HT-2 and T-2 concentrations were significantly correlated with infestation of F. langsethiae (r = 0.65 and r = 0.60, respectively). Concentrations of DON were significantly correlated with F. graminearum infestation (r = 0.61). Furthermore, nivalenol (NIV) was significantly correlated with infestation of F. poae (r = 0.55) and MON with F. avenaceum (r = 0.37). As lower Fusarium infestation and mycotoxin levels were found in organic cereals, factors related to agricultural practice may reduce the risk of contamination with Fusarium mycotoxins. Studies of these issues will be presented separately.

Effect of dietary selenium and omega-3 fatty acids on muscle composition and quality in broilers

BackgroundHuman health may be improved if dietary intakes of selenium and omega-3 fatty acids are increased. Consumption of broiler meat is increasing, and the meat content of selenium and omega-3 fatty acids are affected by the composition of broiler feed. A two-way analyses of variance was used to study the effect of feed containing omega-3 rich plant oils and selenium enriched yeast on broiler meat composition, antioxidation- and sensory parameters. Four different wheat-based dietary treatments supplemented with 5% rapeseed oil or 4% rapeseed oil plus 1% linseed oil, and either 0.50 mg selenium or 0.84 mg selenium (organic form) per kg diet was fed to newly hatched broilers for 22 days.ResultsThe different dietary treatments gave distinct different concentrations of selenium and fatty acids in thigh muscle; one percent linseed oil in the diet increased the concentration of the omega-3 fatty acids 18:3, 20:5 and 22:5, and 0.84 mg selenium per kg diet gave muscle selenium concentration at the same level as is in fish muscle (0.39 mg/kg muscle). The high selenium intake also resulted in increased concentration of the long-chain omega-3 fatty acids EPA (20:5), DPA (22:5) and DHA (22:6), thus it may be speculated if high dietary selenium might have a role in increasing the concentration of EPA, DPA and DHA in tissues after intake of plant oils contning omega-3 fatty acids.ConclusionModerate modifications of broiler feed may give a healthier broiler meat, having increased content of selenium and omega-3 fatty acids. High intakes of selenium (organic form) may increase the concentration of very long-chain omega-3 fatty acids in muscle.

Levels of selected individual polychlorinated biphenyls in different tissues of harbour seals (phoca vitulina) from the Southern coast of Norway.

Concentrations of individual polychlorinated biphenyls (PCBs) were determined in blubber, liver, kidney and brain tissues of 10 male harbour seals (Phoca vitulina) of different ages. The animals were found dead or dying on the southern coast of Norway in 1988 during the morbilli virus epizootic. Twenty-three PCB congeners were investigated in all tissues by capillary gas chromatography with electron capture detection and comparison with standards of the respective congeners (IUPAC numbers 28, 52, 74, 99, 101, 105, 110, 114, 118, 128, 138, 141, 149, 153, 156, 157, 170, 180, 183, 187, 194, 206 and 209). In addition, three toxic coplanar congeners (IUPAC numbers 77, 126 and 169) were investigated in the blubber samples. Total PCB (sum of the determined congeners) in kidney (12-60 nmol g(-1)), liver (12-90 nmol g(-1)) and blubber (10-79 nmol g(-1)) were similar on extractable lipid basis, while total PCB in brain lipid was about 1/10 (1.6-12 nmol g(-1)). The absolute concentrations of the individual congeners in brain lipid correlated poorly with the respective concentrations in each of the other tissues, while the individual congener concentrations in the lipid fractions of kidney, liver and blubber tended to correlate with each other. The relative concentrations of the sum of the most persistent congeners (PCB-99, -128, -138, -153, -170, -180, -187 and -194) correlated in all tissues, and an increase of the relative concentrations of these congeners, most distinct in blubber, with higher carcass weight, was shown. Also, total PCB in blubber correlated with carcass weight. When comparing the relative concentration patterns of individual congeners (per cent of total) in the four tissues, six congeners differed significantly between tissues, and the relative brain concentrations showed most deviation from the other tissues. The concentration of the coplanar PCBs was 1 : 1000-1 : 1000 of the total PCB concentration. The results demonstrate a lower accumulation together with a different concentration pattern in the brain as compared to other tissues.

Influence of agronomic and climatic factors on Fusarium infestation and mycotoxin contamination of cereals in Norway

A total of 602 samples of organically and conventionally grown barley, oats and wheat was collected at grain harvest during 2002–2004 in Norway. Organic and conventional samples were comparable pairs regarding cereal species, growing site and harvest time, and were analysed for Fusarium mould and mycotoxins. Agronomic and climatic factors explained 10–30% of the variation in Fusarium species and mycotoxins. Significantly lower Fusarium infestation and concentrations of important mycotoxins were found in the organic cereals. The mycotoxins deoxynivalenol (DON) and HT-2 toxin (HT-2) constitute the main risk for human and animal health in Norwegian cereals. The impacts of various agronomic and climatic factors on DON and HT-2 as well as on their main producers F. graminearum and F. langsethiae and on total Fusarium were tested by multivariate statistics. Crop rotation with non-cereals was found to reduce all investigated characteristics significantly – mycotoxin concentrations as well as various Fusarium infestations. No use of mineral fertilisers and herbicides was also found to decrease F. graminearum, whereas lodged fields increased the occurrence of this species. No use of herbicides was also found to decrease F. langsethiae, but for this species the occurrence was lower in lodged fields. Total Fusarium infestation was decreased with no use of fungicides or mineral fertilisers, and with crop rotation, as well as by using herbicides and increased by lodged fields. Clay and to some extent silty soils seemed to reduce F. graminearum in comparison with sandy soils. Concerning climate factors, low temperature before grain harvest was found to increase DON; and high air humidity before harvest to increase HT-2. F. graminearum was negatively correlated with precipitation in July but correlated with air humidity before harvest. F. langsethiae was correlated with temperature in July. Total Fusarium increased with increasing precipitation in July. Organic cereal farmers have fewer cereal intense rotations than conventional farmers. Further, organic farmers do not apply mineral fertiliser or pesticides (fungicides, herbicides or insecticides), and have less problem with lodged fields. The study showed that these agronomic factors were related to the infestation of Fusarium species and the concentration of mycotoxins. Hence, it is reasonable to conclude that farming system (organic versus conventional) impacts Fusarium infestation, and that organic management tends to reduce Fusarium and mycotoxins. However, Fusarium infestation and mycotoxin concentrations may be influenced by a range of factors not studied here, such as local topography and more local climate, as well as cereal species and variety.

Response of hepatic xenobiotic metabolizing enzymes in rainbow trout (Oncorhynchus mykiss) and cod (Gadus morhua) to 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD)

Abstract 2,3,7,8-Tetrachlorodibenzo- p -dioxin (2,3,7,8-TCDD) was administered intragastrically twice with 4 days interval, to juvenile cod and rainbow trout, total dose 8 μg/kg body weight. Fish were killed after 9 and 17 days and the effects on hepatic xenobiotic metabolizing enzymes were determined by examining aldrin epoxidase (AE), glutathione-S-transferase (GST) against 1-chloro-2,4-dinitrobenzene (CDNB) and the cytochrome P450-dependent ethoxyresorufin-O-deethylase (EROD) activities, and by immunoquantitating cytochrome P4501A1 using an indirect enzyme-linked immunosorbent assay (ELISA). AE and GST activities were not induced. However, 2,3,7,8-TCDD significantly induced EROD activities in rainbow trout and cod to 1450% and 415%, respectively, of the corresponding controls 9 days after the first treatment. The increase in EROD activity was supported by induction of the main catalyst P4501A1 as revealed by ELISA analyses. The distribution pattern of 14 C-labelled 2,3,7,8-TCDD was studied by whole-body autoradiography. The liver concentration of radiolabelled compound in cod exceeded that of the rainbow trout. However, the degree of hepatic enzyme induction did not correspond to the concentration of radiolabelled 2,3,7,8-TCDD in the liver. Despite of the substantially higher level of 2,3,7,8-TCDD in the liver of cod, the EROD induction was lower in this species compared to rainbow trout.