Low temperature bioremediation of oil-contaminated soil using biostimulation and bioaugmentation with a Pseudomonas sp. from maritime Antarctica.

AIMS: To identify native Antarctic bacteria capable of oil degradation at low temperatures. METHODS AND RESULTS: Oil contaminated and pristine soils from Signy Island (South Orkney Islands, Antarctica) were examined for bacteria capable of oil degradation at low temperatures. Of the 300 isolates cultured, Pseudomonas strain ST41 grew on the widest range of hydrocarbons at 4 degrees C. ST41 was used in microcosm studies of low temperature bioremediation of oil-contaminated soils. Microcosm experiments showed that at 4 degrees C the levels of oil degradation increased, relative to the controls, with (i) the addition of ST41 to the existing soil microbial population (bioaugmentation), (ii) the addition of nutrients (biostimulation) and to the greatest extent with (iii) a combination of both treatments (bioaugmentation and biostimulation). Addition of water to oil contaminated soil (hydration) also enhanced oil degradation, although less than the other treatments. Analysis of the dominant species in the microcosms after 12 weeks, using temporal temperature gradient gel electrophoresis, showed Pseudomonas species to be the dominant soil bacteria in both bioaugmented and biostimulated microcosms. CONCLUSIONS: Addition of water and nutrients may enhance oil degradation through the biostimulation of indigenous oil-degrading microbial populations within the soil. However, bioaugmentation with Antarctic bacteria capable of efficient low temperature hydrocarbon degradation may enhance the rate of bioremediation if applied soon after the spill. SIGNIFICANCE AND IMPACT OF THE STUDY: In the future, native soil bacteria could be of use in bioremediation technologies in Antarctica.

The life cycle of Paraquimperia tenerrima: a parasite of the European eel Anguilla anguilla.

Previous studies on the life history of the nematode eel specialist Paraquimperia tenerrima (Nematoda: Quimperiidae) have failed to determine whether an intermediate host is required in the life cycle. In the laboratory, eggs failed to hatch below 10 degrees C, hatching occurring only at temperatures between 11 and 30 degrees C. Survival of the free-living second stage larvae (L2) was also temperature dependent, with maximal survival between 10 and 20 degrees C. Total survival of the free-living stages (eggs and L2) is unlikely to exceed a month at normal summer water temperatures, confirming that parasite could not survive the 6 month gap between shedding of eggs in spring and infection of eels in early winter outside of a host. Eels could not be infected directly with L2, nor could a range of common freshwater invertebrate species. Third stage larvae (L3) resembling P. tenerrima were found frequently and abundantly in the swimbladder of minnows Phoxinus phoxinus from several localities throughout the year and were able to survive in this host in the laboratory for at least 6 months. Third stage larvae identical to these larvae were recovered from minnows experimentally fed L2 of P. tenerrima, and eels infected experimentally with naturally and experimentally infected minnows were found to harbour fourth stage larvae (L4) and juvenile P. tenerrima in their intestines. Finally, the whole life cycle from eggs to adult was completed in the laboratory, confirming that minnows are an obligate intermediate host for P. tenerrima.

Accounting for differences in estrogenic responses in rainbow trout (Oncorhynchus mykiss: Salmonidae) and roach (Rutilus rutilus: Cyprinidae) exposed to effluents from wastewater treatment works.

Effluents from wastewater treatment works (WwTWs) contain estrogenic substances that induce feminizing effects in fish, including vitellogenin (VTG) synthesis and gonadal intersex. Fish vary in their responsiveness to estrogenic effluents, but the physiological basis for these differences are not known. In this study, uptake of estrogen from two WwTW effluents (measured in hydrolyzed bile) and estrogenic response (VTG induction) were compared in a salmonid (rainbow trout, Onchorhynchus mykiss) and a cyprinid fish (roach, Rutilus rutilus). Immature rainbow trout were more responsive than maturing roach to the estrogenic effluents. The more potent of the two estrogenic effluents (containing between 24.3 and 104.1 ng estradiol-17beta equivalents/L [E2eq/L]) resulted in a 700-fold and 240-fold induction of plasma VTG in male and female trout, respectively, but only a 4-fold induction in roach (and in males only). The less potent effluent (varying between 4.1 and 6.8 ng E2eq/L) induced VTG in the trout only, with a 4-fold and 18-fold induction in males and females, respectively. In fish exposed to tap water, the estrogenicity of the hydrolyzed bile was 0.03+/-0.01 ng E2eq/microL (for both sexes in trout), 0.18+/-0.04 ng E2eq/microL in male roach, and 0.88+/-0.15 ng E2eq/microL in female roach. The higher bile content of estrogen in control roach reflected their more advanced sexual status (and thus higher endogenous estrogen) compared with the immature female trout. In trout maintained in effluents, the bile content of estrogen was 100-fold and 30-fold higher than controls at WwTW A and B, respectively. Bioconcentration factors (BCFs) for estrogenic activity in bile were between 16 344 and 46 134 in trout and between 3543 and 60 192 in roach (no gender differences were apparent). There were strong correlations between VTG induction and the estrogenic activity of bile extracts for both trout and roach. The results confirm that estrogenic contaminants bioconcentrate to a high degree in fish bile and that the level (and nature) of this accumulation may accountfor responsiveness to the endocrine disruptive effects of estrogenic effluents. Immature fish were the more appropriate life stage for quantifying estrogen exposure and uptake in bile, as they contain little circulating endogenous oestrogen compared with sexual maturing fish. The nature of the estrogenic contaminants is detailed in an accompanying paper.

Androgenic and estrogenic effects of the synthetic androgen 17alpha-methyltestosterone on sexual development and reproductive performance in the fathead minnow (Pimephales promelas) determined using the gonadal recrudescence assay.

The effects of the androgen, 17alpha-methyltestosterone were assessed on sexual development and reproductive performance in the fathead minnow (Pimephales promelas) using a gonadal recrudescence assay. In this assay, mature male and female fathead minnow, previously kept under simulated winter conditions (15 degrees C; 8:16 h light:dark regime) were transferred to simulated summer conditions (25 degrees C water temperature; 16:8 h light:dark regime) to induce gonadal recrudescence. To assess sexual development fish were exposed to nominal concentrations of 0, 0.1, 1, 5 and 50 microg/L 17alpha-methyltestosterone. After 3 weeks of chemical exposure, effects on condition (condition factor, CF), plasma vitellogenin (VTG), secondary sex characteristics, gonad growth (gonadosomatic index; GSI) and gonad histology were investigated. Reproductive performance, including reproductive output (egg production), spawning behaviour, and fertilisation rate were measured over a subsequent 3-week-period in breeding adults maintained in clean water. 17alpha-Methyltestosterone had no effects on the condition of fish at any of the doses tested. 17alpha-Methyltestosterone induced both androgenic and estrogenic effects with females generally more affected by 17alpha-methyltestosterone than males: atretic follicles and male-specific sex characteristics (androgenic effect) were induced in females at > or = 0.1 and > or = 1 microg/L 17alpha-methyltestosterone, respectively. An inhibitory effect on ovary growth occurred at an exposure concentration of 50 microg/L 17alpha-methyltestosterone. In males 1 microg/L 17alpha-methyltestosterone induced a concentration-response induction of plasma vitellogenin (estrogenic effect) likely due to its conversion into 17alpha-methylestradiol, rather to the competition with endogenous steroids and their cross reactivity with the estrogen receptor. In the fish breeding studies, concentration-dependent reductions in egg number, fertilisation rate and increases in abnormal sexual behaviour in females were observed. All of these effects occurred at exposure concentrations of > or = 5 microg/L 17alpha-methyltestosterone. Thus, it could be assumed that the observed estrogenic effects in male fathead minnow were likely to the conversion of 17alpha-methyltestosterone into the estrogen 17alpha-methylestradiol, rather to the acting of 17alpha-methyltestosterone itself. In conclusion to this, showing hormonally activity of 17alpha-methyltestosterone in fish down to 100 ng/L, indicates that its potency was close to the range of several naturally occurring estrogens.

Atmospheric distribution of polychlorinated dibenzo-p-dioxins, dibenzofurans (PCDD/Fs), and non-ortho biphenyls (PCBs) along a North-South Atlantic transect.

Air samples were taken on board the RRS Bransfield (typically for 24-72 h), during an Atlantic cruise from the U.K. to Antarctica in October-December 1998, to investigate the global scale distribution of PCDD/Fs and coplanar PCBs. Highest concentrations of Cl2-8DD/Fs all occurred between 25 and 52 degrees N; lowest concentrations were measured around approximately 60 degrees S and further south. Cl3DFs showed highest overall concentrations (up to 9800 fg/m3), followed by Cl2DFs (up to 5300 fg/m3) and OCDD (up to 1300 fg/m3). Lowest concentrations, measured in the remote Southern hemisphere, were generally 2 orders of magnitude lower than their highest concentrations over the North Atlantic. Concentrations of PCB-77 were higher in the northern hemisphere, while PCB-126 and PCB-169 exhibited highest concentrations around the equator. Evidence was obtained for substantial emissions of PCDD/Fs off west Africa and while in the port of Montevideo, Uruguay. Shifts in PCDD/F profile distribution were observed on increasing distance from source regions, such that those from the most remote locations were dominated by Cl3DF (approximately 40% of the total) and OCDD (approximately 20%). Gas-particle partition data was obtained for all samples. Cl4-6DD/Fs showed the widest range, varying between 10 and 90% of the total in the gas phase, depending on location/ temperature. The study gave limited evidence for the influence of OH-radical initiated depletion reactions of gaseous PCDD/Fs. The global atmospheric burden is estimated to be on the order of 350 kg sigmaCl4-8DD/Fs and approximately 3 kg sigmaTEQ.

Long-term dynamics of Ligula intestinalis and roach Rutilus rutilus: a study of three epizootic cycles over thirty-one years.

Data are presented on 2 full epizootic cycles and the start of a third of Ligula intestinalis in roach Rutilus rutilus in a small lake, and the relationships of these cycles to the densities of rudd, Scardinius erythrophthalmus, and Great Crested Grebes, Podiceps cristatus, over 31 years. The parasite was introduced to the lake by P. cristatus in 1973 at a time when the roach population had increased in response to eutrophication to a level at which individual fish growth was stunted and the hithero dominant rudd population had declined in numbers as a consequence of inter-specific competition with roach. Ligula prevalence peaked at 28% in only 2 years: thereafter parasite-induced host mortality caused a decline in the roach population, releasing fish from stunting and allowing the rudd population to recover. The consequent improved growth of roach individuals and their short life-span reduced Ligula transmission rates and prevalence levels declined to approximately 1% although Ligula nevertheless persisted for a further 10 years. Following a massive winter-kill of the fish populations in 1984-1985, fish and Ligula numbers declined to barely detectable levels and the parasite disappeared from samples. Rudd recovered first, then roach and interspecific competition again led to a decline in rudd numbers. This increase in roach numbers led to a decrease in roach growth rates, which coincided with the re-colonization of the lake by Ligula. This second epizootic of Ligula peaked within 2 years in 1991-1992, when up to 78% of roach were infected with a maximum abundance of 2.2 parasites and intensity of 21 parasites. Heavy parasite-induced mortality of roach led to a decline in numbers, an improvement in individual growth rate and a reduction of Ligula transmission rates such that the epizootic died out in 1996. Similar conditions of roach numbers and growth prevailed at the start of a third cycle in 1998. The course of events over the second cycle was so similar to that of the first that it confirms the interpretations of that cycle. Comparison with other localities shows that epizootics of Ligula always coincide with rapid increases in roach numbers, for whatever cause, and stunted growth, which together attract piscivorous birds. At the start of a cycle Ligula is a major determinant of the population dynamics of the roach, but at the end of the cycle the fish population dynamics determine those of the parasite. The cycles are not regulated and the roach-Ligula system is inherently unstable.

The significance of PCBs in the atmosphere of the southern hemisphere.

Air monitoring stations were set up at 2 sites in the southern hemisphere--Moody Brook, Falkland Island (51 degrees 25' S, 57 degrees 56' W) and Halley, Research Station, Antarctica (75 degrees 35' S, 26 degrees 30' W). PCBs were monitored at the stations throughout 1999. Highest concentrations were observed when temperatures were greater. In general, concentrations were greater at Moody Brook than at Halley, although the difference in concentrations between sites was less for more chlorinated congeners. Air concentrations at both sites were compared with samples collected nearby over-water. Over water air concentrations were found to be greater than over land air concentrations. Concentrations were also compared with literature data for air concentrations at a remote site in the Canadian Arctic. Atmospheric concentrations of tri-chlorinated biphenyls were found to be approximately double those reported for Ellesmere Island in the Canadian Arctic, whilst concentrations in samples from Antarctica were very similar to those found in the high Arctic. Most other PCBs were a factor of 2-4 greater in the Canadian Arctic.

Intra-atrial smoke-like echoes and thrombi formation.

Between December 1980 and December 1987, approximately 6,000 two-dimensional echocardiographic examinations were performed in our noninvasive cardiac laboratory. During this period, five cases of intra-atrial smoke-like echoes were diagnosed. The four patients with dense smoke-like echoes in the left atria all had echocardiographic evidence of severe mitral stenosis and thrombi in the left atrial cavity, both of which were confirmed at open-heart surgery. One patient with right-sided cardiomyopathy had echocardiographic evidence of dense smoke-like echoes and thrombus in the right atrial cavity. One of our five patients was receiving anticoagulant therapy, and none had a history of thromboembolism. We conclude that using conventional two-dimensional echocardiographic techniques, smoke-like echoes in the atria are a very infrequent finding and are very commonly associated with formation of thrombi.

Spectroscopic evidence for a photosensitive oxygenated state of ammonia mono-oxygenase.

Photoinactivation of ammonia oxidation by Nitrosomonas europaea cells by near-u.v. light was confirmed and further shown to occur with the same rate constant as loss of bromoethane-oxidation activity. Hydroxylamine oxidation was much less photosensitive. Protection against inactivation was afforded by anaerobiosis, organic substrates of ammonia mono-oxygenase such as bromoethane, or metal-ion-chelating agents such as thiourea. The presence of 10 mM-NH4+ or 1 mM-hydroxylamine made little difference, whereas hydrazine had a potentiating effect. Illumination of cells also caused a bleaching in the absorption spectrum around 380 nm, along with changes in the cytochrome gamma-band region. Similar effects below 400 nm were obtained when organic substrates and inhibitors of the mono-oxygenase were added to cells in the dark. The copper proteins haemocyanin and tyrosinase have a photosensitive oxygenated state with a near-u.v. absorption band of similar half-width. They also have a sensitivity to chelating agents similar to that of ammonia mono-oxygenase. The experimental results are explained in terms of a three-stage catalytic cycle analogous to that for tyrosinase. In resting cells most of the enzyme is believed to be in an oxygenated (Oxy) form, which absorbs maximally at 378 nm and is photosensitive. In the presence of a substrate, one O atom is inserted into the substrate and the other is reduced to water, leaving the enzyme in an oxidized (Met) state. This is followed by a two-electron reduction of the proposed binuclear copper site to give a reduced (Deoxy) state, which can bind O2 to complete the cycle.