Distribution, fate and risk assessment of antibiotics in sewage treatment plants in Hong Kong, South China.

Occurrence, removal, consumption and environmental risks of sixteen antibiotics were investigated in several sewage treatment plants (STPs) featuring different treatment levels in Hong Kong, China. Cefalexin, ofloxacin and erythromycin-H(2)O were predominant with concentrations of 1020-5640, 142-7900 and 243-4740 ng/L in influent, respectively; their mass loads were comparable to levels reported in urban regions in China and were at the high end of the range reported for western countries. The target antibiotics behaved differently depending on the treatment level employed at the STPs and relatively higher removal efficiencies (>70%) were observed for cefalexin, cefotaxime, amoxicillin, sulfamethoxazole and chloramphenicol during secondary treatment. ß-lactams were especially susceptible to removal via the activated sludge process while macrolides were recalcitrant (<20%) in the dissolved phase. Two fluoroquinolones, ofloxacin (4%) and norfloxacin (52%), differed greatly in their removal efficiencies, probably because of disparities in their pK(a) values which resulted in different sorption behaviour in sludge. Overall antibiotic consumption in Hong Kong was back-calculated based on influent mass flows and compared with available prescription and usage data. This model was verified by a good approximation of 82% and 141% to the predicted consumption of total ofloxacin, but a less accurate estimate was obtained for erythromycin usage. Risk assessment indicated that algae are susceptible to the environmental concentrations of amoxicillin as well as the mixture of the nine detected antibiotics in receiving surface waters.

Brown mussels (Perna perna) and semi-permeable membrane devices (SPMDs) as indicators of organic pollutants in the South African marine environment.

A distinct lack of historical and current data on the status of organic pollutant contaminants within the South African marine environment is evident. This has highlighted the need for more current organic pollutant assessments. Reference mussels and SPMDs were transplanted at five South African harbour sites to assess organic bioaccumulation in brown mussels (Perna perna) and semi-permeable membrane devices (SPMDs). Spatial patterns of PAH and PCB contaminants were determined by GC-MS and GC-ECD after appropriate sample preparation. Significant (p<0.05) spatial differences were observed between the sites. Results indicate no correlations between the passive device and the transplanted mussels; however the SPMDs provided complementary information on the presence of dioxin-like PCBs within the environment not detected by the mussel. The results indicate that information provided by both the mussels and SPMDs allow for a more in depth scrutiny of environmental conditions as a result of anthropogenic influence.

Application of artificial mussels (AMs) under South African marine conditions: a validation study.

Over the last three decades there has been a significant decline in the number of marine pollution monitoring-related studies in South Africa. Thus, the current study was conducted to assess the current state of metal contamination within the South African marine environment through the validation of the artificial mussel (AM). Indigenous reference mussels (Perna perna) were deployed alongside the passive device within the South African marine environment for a 6 week exposure period. Analysis of metal uptake (Cd, Cr, Cu, Pb and Zn) was determined by filtration and elution of the AM chelex resins, microwave digestion of the transplanted mussels, and determination of their metal concentrations by ICP-MS and ICP-OES analysis. Uptake patterns between the AM and transplanted mussels showed significant temporal and spatial correlation for the majority of the elements analysed. While the AM provided relevant and complementary information on the dissolved metal concentrations, limitations were also observed.

Alpha-particle radiobiological experiments using thin CR-39 detectors.

The present paper studied the feasibility of applying comet assay to evaluate the DNA damage in individual HeLa cervix cancer cells after alpha-particle irradiation. We prepared thin CR-39 detectors (<20 microm) as cell-culture substrates, with UV irradiation to shorten the track formation time. After irradiation of the HeLa cells by alpha particles, the tracks on the underside of the CR-39 detector were developed by chemical etching in (while floating on) a 14 N KOH solution at 37 degrees C. Comet assay was then applied. Diffusion of DNA out of the cells could be generally observed from the images of stained DNA. The alpha-particle tracks corresponding to the comets developed on the underside of the CR-39 detectors could also be observed by just changing the focal plane of the confocal microscope.

Trace organic contamination in biota collected from the Pearl River Estuary, China: a preliminary risk assessment.

The marine ecosystem of the Pearl River Delta, located on the southern coast of China, has been heavily exploited following the rapid economic growth that has occurred since the 1980s. This investigation aimed to elucidate trace organic contamination in marine biota inhabiting the Pearl River Delta area. Biota samples, including green-lipped mussels (Perna viridis), oysters (Crassostrea rivularis) and shrimp (Penaeus orientalis) were sampled from 16 stations fringing the Estuary. Elevated concentrations (on a dry weight basis) of polycyclic aromatic hydrocarbons (27.8-1041.0 ng/g), petroleum hydrocarbons (1.7-2345.4 microg/g), polychlorinated biphenyls (2.1-108.8 ng/g), DDTs (1.9-79.0 ng/g), and hexachlorocyclohexanes (n.d.-38.4 ng/g) were recorded. A human health risk assessment was conducted to estimate the risk to local residents associated with the consumption of biota collected from the Pearl River Estuary. The results indicated that PCBs were at levels that may cause deleterious health effects in populations that consume large amounts of seafood. However, it would be instructive to establish health criteria for trace organic contaminants that are specific to the local populations, in order to derive a more accurate and relevant health risk assessment.

A preliminary risk assessment of organochlorines accumulated in fish to the Indo-Pacific humpback dolphin (Sousa chinensis) in the Northwestern waters of Hong Kong.

The Indo-Pacific humpback dolphin is considered threatened due to several factors including pollution in Hong Kong and the risks due to consumption of fish tainted with polychlorinated biphenyls (PCBs) and organochlorine pesticides were assessed. Six species of fish Collichthys lucida, Pseudosciaena crocea, Johnius sp., Thryssa sp., Mugil sp. and Trichiurus sp., which comprise the main prey species of humpback dolphins were collected for analyses. Risks due to total PCBs, total TEQs, PCB 118 and the pesticides were assessed with the use of toxicity reference values as the threshold reference benchmarks. The calculated risk quotients (RQs) showed that the risks associated with organochlorines were generally low. The highest RQ was associated with total TEQs suggesting that dioxin-like PCBs may pose the highest risk to the dolphins. The HCHs, total PCBs and heptachlor had comparatively high RQs and thus they should also be the priority organochlorines that would require further investigation.

A preliminary risk assessment of trace elements accumulated in fish to the Indo-Pacific Humpback dolphin (Sousa chinensis) in the northwestern waters of Hong Kong.

In order to assess the potential risks associated with consumption of contaminated prey items to the Indo-Pacific Humpback dolphin (Sousa chinensis), fish species (Collichthys lucida, Pseudosciaena crocea, Johnius sp., Thryssa sp., Mugil sp. and Trichiurus sp.) representing the main food items of the dolphin were collected from the northwestern waters of Hong Kong, including the Sha Chau and Lung Kwu Chau Marine Park, which form the main habitat of the dolphin in Hong Kong. Within these waters, there are several potential sources of pollution including significant inputs from the Pearl River catchment, several major sewage outfalls and a series of mud pits that receive contaminated dredged sediments. Concentrations of thirteen trace elements (Ag, As, Cd, Co, Cr, Cs, Cu, Hg, Mn, Ni, Se, V, and Zn) in the fish tissue were analyzed by inductively coupled plasma mass spectrometer (ICP-MS). An assessment of the risks of adverse effects on the dolphin due to consumption of tainted fish was undertaken using two toxic reference benchmarks, namely the reference dose (RfD) and toxicity reference value (TRV). The risk quotient (RQ) calculated for each element showed that the risks from consumption of fish were generally low and within safe limits. The risks associated with arsenic, cadmium and mercury were, however, elevated. The highest calculated RQ was associated with total arsenic; however, the majority of arsenic in marine organisms tends to be in the non-toxic organic form, and the actual risk to the dolphin due to this metalloid is likely to be lower.

Application of the comet and micronucleus assays to the detection of B[a]P genotoxicity in haemocytes of the green-lipped mussel (Perna viridis).

Green-lipped mussels (Perna viridis) were exposed to water-borne benzo[a]pyrene (B[a]P) at nominal concentrations of 0, 0.3, 3 and 30 microg l(-1) for up to 12 days, and both the relative levels of DNA strand breaks (assessed using an alkaline comet assay) and the proportion of micronucleus (MN) formation were monitored in mussel haemocytes at days 0, 1, 3, 6 and 12. The results of the comet assay indicated that an increase in the proportion of strand breaks occurred generally with increasing B[a]P concentration, but a significant decrease in the levels of DNA damage was observed after exposure for 12 days at all concentrations tested, suggesting that the patterns of changes in the levels of DNA strand breakage can be explained by the threshold dependent DNA repair theory. Moreover, the relatively slow development and recovery of the DNA damage response in mussel haemocytes in comparison with previous findings utilizing P. viridis hepatopancreas suggests that the response of DNA alteration upon exposure to B[a]P may be tissue-specific in this species. Monitoring the frequency of micronucleus development in mussel haemocytes indicated both dose- and time-response relationships within the exposure period. Furthermore, the levels of DNA strand breakage correlated well with the levels of micronucleus induction, suggesting a possible cause and effect relationship between the two damage types. We suggest that DNA strand breakage and micronucleus formation in mussel haemocytes can potentially be used as convenient biomarkers of exposure to genotoxicants in the marine environment.

Antioxidant responses to benzo[a]pyrene and Aroclor 1254 exposure in the green-lipped mussel, Perna viridis.

In this study, the green-lipped mussel, Perna viridis (L.), was exposed to two concentrations of benzo[a]pyrene (B[a]P) (0.3 microg l(-1); 3 microg l(-1)) and two concentrations of Aroclor 1254 (0.5 microg l(-1); 5 microg l(-1)). In addition, a mixture of the contaminants was used (0.3 microg l(-1) B[a]P+0.5 microg l(-1) Aroclor 1254; 3 microg l(-1) B[a]P+5 microg l(-1) Aroclor 1254). All concentrations were nominal. A suite of enzymes [glutathione S transferase (GST), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR)], glutathione (GSH) level and lipid peroxidation (LPO) in the mussel gill and hepatopancreas were monitored over 18 days. CAT and GSH in gill tissue were positively correlated with concentration of Aroclor 1254. Activity of hepatic GST and SOD was significantly related to body burden of Aroclor 1254. LPO, GR and GPx in gill and hepatopancreas and hepatic GST were positively correlated with B[a]P concentration. The results indicate the importance of using biomarkers specific to the type of contaminant(s) that are likely to be present. Controlled laboratory experiments, such as this study, are useful in ascertaining biomarkers suitable for use with complex contaminant mixtures in the marine environment.

Mussel-based monitoring of trace metal and organic contaminants along the east coast of China using Perna viridis and Mytilus edulis.

Green lipped mussels, Perna viridis, and blue mussels, Mytilus edulis, were collected from seven locations along the east coast of China in September and October 2001. The mussel tissues were analyzed for metals (Ag, As, Cd, Cr, Ni, Pb, Se, Zn, Cu, Fe and Hg), and trace organic contaminants including organochlorine compounds (OCs), polycyclic aromatic hydrocarbons (PAHs; based on 24 individual PAHs), polychlorinated biphenyls (PCBs) and petroleum hydrocarbons (PHCs). Maximum concentrations of Ag, As, Cd, Cr, Ni, Pb, Se, Zn, Cu, Fe and Hg in the mussel tissues were 0.14, 26.76, 5.31, 15.72, 4.78, 2.93, 7.40, 231.0, 54.17, 1002 and 317.3 microg/g dry weight respectively. Levels of DDTs, PAHs, PCBs and PHCs in the mussel samples were 14-640, 456-3495, 1-13 ng/g and 621-2863 microg/g dry weight, respectively. Results of this study indicated that contaminant levels were, in general, higher or at least comparable to those reported in other local or regional studies. Mussel samples collected in Chongming Dao in Shanghai and Jiao Zhou Wan in Qingdao had significantly higher levels of metal and trace organic contaminants among the seven sampling stations. Examination of the contaminant profiles suggests that PHCs originated from petrogenic sources, while both petrogenic and pyrolytic sources were important for PAHs. The generally high levels of metals and organochlorine compounds are probably the result of increasingly intense industrial activities along the east coast of China. An assessment of potential risks to human health due to consumption of the mussels was undertaken for the metals, PCBs, DDTs and chlordanes, and the results indicated that all metals, except Ni, could pose a health risk to heavy seafood consumers, while Hg appears to be of concern even for low level consumers.

Exposure and time dependent DNA strand breakage in hepatopancreas of green-lipped mussels (Perna viridis) exposed to Aroclor 1254, and mixtures of B[a]P and Aroclor 1254.

Green-lipped mussels (Perna viridis) were exposed to Aroclor 1254 (0.5, 5 and 50 microgl(-1)) and a mixture of benzo[a]pyrene (B[a]P) and Aroclor 1254 (0.3+0.5 and 3+5 microgl(-1)) for 12 days. On day 0, 1, 3, 6 and 12, the levels of DNA strand breaks in the mussel hepatopancreas were monitored using an alkaline unwinding assay. The results were compared to the findings of a previous study in which the levels of DNA strand breakage in the same species were measured following exposure to various concentrations of B[a]P (0.3, 3 and 30 microgl(-1)). The results indicated that Aroclor 1254 at a concentration

To fix or not to fix: the role of 2-phenoxyethanol in rabbit, Oryctolagus cuniculus, chin gland secretion.

The European rabbit (Oryctolagus cuniculus) uses the secretion of the chin gland to maintain dominance hierarchies in the wild. Recent work has investigated changes in the secretion when social status is manipulated in the rabbit. When a rabbit becomes dominant, a new compound appears in his secretion, 2-phenoxyethanol. This compound is used as a fixative in the perfume industry. This study investigates whether the compound performs a similar function in the secretion of the rabbit. 2-Phenoxyethanol is not detected olfactorially by rabbits, and slows the release rate of some of the compounds that occur naturally in rabbit chin gland secretion. We suggest that when a rabbit becomes dominant, he adds a fixative to his secretion so that his scent will persist in the environment and not dissipate. He will thus come to dominate the olfactory environment, in much the same way as he does the physical environment.

A rapid population expansion retains genetic diversity within European rabbits in Australia.

The well documented historical translocations of the European rabbit (Oryctolagus cuniculus) offer an excellent framework to test the genetic effects of reductions in effective population size. It has been proposed that rabbits went through an initial bottleneck at the time of their establishment in Australia, as well as multiple founder events during the rabbit's colonization process. To test these hypotheses, genetic variation at seven microsatellite loci was measured in 252 wild rabbits from five populations across Australia. These populations were compared to each other and to data from Europe. No evidence of a genetic bottleneck was observed with the movement of 13 rabbits from Europe to Australia when compared to French data. Within Australia the distribution of genetic diversity did not reflect the suggested pattern of sequential founder effects. In fact, the current pattern of genetic variation in Australia is most likely a result of multiple factors including mutation, genetic drift and geographical differentiation. The absence of reduced genetic diversity is almost certainly a result of the rabbit's rapid population expansion at the time of establishment in Australia. These results highlight the importance of population growth following a demographic bottleneck, which largely determines the severity of genetic loss.

Risk to breeding success of fish-eating Ardeids due to persistent organic contaminants in Hong Kong: evidence from organochlorine compounds in eggs.

Eggs of two Ardeid species, the Little Egret (Egretta garzetta) and the Black-crowned Night Heron (Nycticorax nycticorax), were collected from two egretries located in the New Territories of Hong Kong with one located near the internationally acclaimed wetland reserve, the Mai Po Marshes, and the other in a remote site (A Chau). The eggs were analysed for organochlorine (OC) compounds including the DDTs, PCBs, hexachlorocyclohexanes (HCHs) and the chlordanes (CHLs). All of the OCs under investigation were detected in the eggs of both species with significantly higher levels in the Little Egret (DDTs, 560-2200; PCBs, 270-1700; CHLs, 81-470 ng g(-1) wet weight) than the Night Heron (DDTs, 210-1200; PCBs, 85-600; CHLs 59-75 ng g(-1) wet weight). The DDTs consisted mainly of DDE with levels ranging from 85% to 95% of the total. The HCHs were at about the same levels in both species (8.4-30 ng g(-1) wet weight). All of the OCs had linear concentration probability distributions on a log-normal basis which were used to evaluate exposure associated with these compounds as part of a probabilistic risk analysis. A linear dose/response relationship for the percentage reduction in the survival of young associated with DDE in eggs was developed. This probabilistic relationship was used to establish the threshold level (1000 ng g(-1) wet weight) at which there was a significant level of reduction in the survival of young above zero and the variability in DDE concentrations at this effect level. Using a threshold level of 1000 ng g(-1), the calculated Risk Quotient (RQ) had a 12.4% probability of RQ exceeding unity with the Night Heron, and 40.9% with the Little Egret. These results indicate that the DDTs in eggs would be expected to be associated with adverse effects on the survival of young of both species, particularly the Little Egret.

Relationships between tissue concentrations of chlorinated hydrocarbons (polychlorinated biphenyls and chlorinated pesticides) and antioxidative responses of marine mussels, Perna viridis.

Marine mussels, Perna viridis, were transplanted from a reference site to various polluted sites around Hong Kong. After 30 d of exposure, antioxidative responses in the gills and hepatopancreas and tissue concentrations of chlorinated hydrocarbons [polychlorinated biphenyls (PCBs) and chlorinated pesticides (CPs)] were determined for individual mussels. Glutathione S transferase (GST) and glutathione (GSH) were positively correlated with tissue PCB concentrations. Only one of the enzymatic antioxidants, glutathione peroxidase (GPx), showed significant response to tissue PCB. No significant correlation was found between tissue concentrations of chlorinated hydrocarbons and other enzymatic antioxidants (superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR) and NADPH DT-diaphorase (DT-d). Oxidative stress, measured as thiobarbituric acid reactive substances, was correlated with chlorinated pesticide concentrations in tissues. This study demonstrated a correlation between GST/ GSH and chlorinated hydrocarbons. The apparent lack of correlation between trace organic pollutants and some of the enzymatic antioxidants may be due to the inhibitory effects caused by these chemicals. The above results suggest that more investigations are needed before these enzymes can be used as biomarkers.

A study of the biological partitioning behavior of n-alkanes and n-alkanols in causing anesthetic effects.

n-Alkanes and n-alkanols are two groups of common volatile organic compounds (VOCs) having potential anesthetic effects on workers and building occupants. A partition model based on the octanol-air partition coefficient was developed in this investigation to describe the biological partitioning of n-alkanes and n-alkanols in causing general anesthesia. Data on anesthetic potency (minimum alveolar concentration, MAC) for the test groups in rats were found to fit the model. The slight difference between the n-alkanes and n-alkanols in testing the model could be largely eliminated by correcting for the potential partial pressure gradients of the long-chain alkanes across the blood-brain barrier. The corrected MAC data for the two test groups fit well onto one common activity-partition regression line. This suggests that n-alkanes and n-alkanols may share a common biophase or mechanistic pathway for anesthesia. The present findings may provide some useful insight into setting anesthesia-based health standards for VOC mixtures.

Semiochemicals and social signaling in the wild European rabbit in Australia: I. Scent profiles of chin gland secretion from the field.

The European rabbit (Oryctolagus cuniculus) uses the secretion of the chin gland in the maintenance of social status. Previous work has concentrated on secretion collected directly from the animal. In this study, the analysis was conducted by collecting scent marks made by free-ranging animals. Scent marks were found to be concentrated at the center of the area controlled by a social group, and at the boundaries between two adjacent social groups. Only the mark from dominant animals could be identified. Marks were also collected from the skin of rabbits, where they had been placed by the dominant individual. The mark found on the head of a subordinate animal may, in the future, be used to identify the dominant animal of the social group, who placed the mark.

Risk to breeding success of Ardeids by contaminants in Hong Kong: evidence from trace metals in feathers.

The feathers of two Ardeid species, the Little Egret (Egretta garzetta) and the Black-crowned Night Heron (Nycticorax nycticorax) were collected from six egretries and two egretries respectively, located in different areas in the New Territories of Hong Kong, including the Mai Po Marshes (within a Ramsar site). These feathers were digested and concentrations (microg/g dry weight) of copper (4.6-19.4), iron (8.1-641.3), manganese (0.4-19.4), zinc (51.3-183.5), lead (0.1-5.1), cadmium (0.01-0.15), chromium (0.06-1.7) and mercury (0.0-7.1) were determined by ICP-AES, ICP-MS and CVAAS. The levels of manganese, mercury and lead found were equal to or less than the concentrations found in previous investigations, reflecting a slight downward trend most apparent with lead. As a general rule, the levels of lead and mercury were higher in the egretries close to the polluted Deep Bay. A probabilistic risk assessment of the possible adverse effects on the breeding success of the Little Egret was carried out with respect to mercury, lead and cadmium. It was concluded that mercury (0.5-7.1 microg/g dry weight feathers) probably has had adverse effects at the Au Tau egretry of the Little Egrets, but there was no evidence of adverse effects at other egretries. The probabilistic analysis also indicated a low likelihood of adverse effects of mercury on the breeding of the Black-crowned Night Herons at A Chau (0.3-1.2 microg/g) and Mai Po Village (0.0-1.4 microg/g). The evidence for the effects of lead and cadmium was limited but suggested there may possibly be adverse effects with lead but not cadmium.

Semiochemicals and social signaling in the wild European rabbit in Australia: II. Variations in chemical composition of chin gland secretion across sampling sites.

The volatile components of the chin gland secretion of the wild European rabbit, Oryctolagus cuniculus (L.), were investigated with the use of gas chromatography. Studies of the chemical nature of this secretion by previous workers demonstrated that it was important in the maintenance of social structure in this species. This study identified 34 different volatile components that consist primarily of aromatic and aliphatic hydrocarbons. Especially common are a series of alkyl-substituted benzene derivatives that provide most of the compound diversity in the secretion. Samples of chin gland secretion collected from animals at three different geographical locations, separated by more than 100 km, showed significant differences in composition. This work suggests that variation among populations needs to be considered when undertaking semiochemical research. Alternate nonparametric methods are also used for the analysis of chromatographic data.

A comparison of mussels (Perna viridis) and semi-permeable membrane devices (SPMDs) for monitoring chlorinated trace organic contaminants in Hong Kong coastal waters.

A comparison of mussels (Perna viridis) and semi-permeable membrane devices (SPMDs) was carried out at five sites, representing a gradient of contaminant concentrations, in Hong Kong coastal waters. Mussels, originally collected from a "clean" location, were deployed along with SPMDs at each site for 30 days. Analyses for chlorinated pesticides and polychlorinated biphenyls (PCBs) indicated that SPMDs have potential as monitoring tools, and to some extent can overcome the problems associated with mussels, such as natural variability, differing age, sex, and physical condition. However, in most cases, SPMDs failed to rank the sites in the same order as mussels in terms of contaminant concentrations. Nonetheless, in localities where mussels cannot survive--as shown at Kwun Tong in the present experiment--SPMDs may be valuable in providing an indication of potentially bio-available lipophilic pollutants.