The effects of landscape-level disturbance on the composition of Minnesota caddisfly (Insecta: Trichoptera) trophic functional groups: evidence for ecosystem homogenization

Over 300,000 caddisfly specimens representing 249 species were collected from nearly 250 sites throughout Minnesota during 2000 and 2001 to determine the effects of human disturbance on the composition of caddisfly trophic functional groups at the landscape level. Canonical correspondence analysis determined that stream width was the most important variable influencing functional group composition in regions of the state with relatively low disturbance, and that differences in the caddisfly fauna between sizes of streams generally followed trends predicted by the river continuum concept. In regions of the state with moderate disturbance, both stream width and the percentage of disturbed habitat upstream of a site were important variables influencing functional group composition. In highly disturbed regions, no variables corresponded to changes in the composition of caddisfly functional groups. Instead, ecosystems were homogeneous: fine-particle filtering collectors dominated in all sizes of streams. The observed aquatic ecosystem homogenization is attributed mostly to input of fine-particle organic and inorganic sediment from extensive agriculture.     

Organochlorine residual concentrations in cattle egret from the Punjab Province, Pakistan

In this study, residual concentration of organochlorine pesticides (OCPs) in the sediments, prey, and eggs of Bubulcus ibis were measured from three breeding heronries from the Punjab province of Pakistan. Pattern of contamination in eggs followed the order: DDTs > HCHs > heptachlor > aldrin. Overall, pesticide residual concentrations were greater in eggs of cattle egrets collected from heronry on the River Ravi. Among HCHs, ??-HCH was more prevalent in eggs, whereas DDTs followed the order: DDD > DDE > p,p ^??-DDT > o,p ^??-DDT. Eggshell thinning was detected which showed negative relationship with residual concentration of DDE. In prey samples, residual concentration of POPs followed the order: DDTs > HCHs > dicofol > heptachlor; however, contamination pattern in sediments followed a slightly different order: DDTs > heptachlor > dicofol > HCHs > dieldrin > aldrin. Concentration of ??-HCH was more prevalent in sediments and comparatively greater concentrations of POPs were measured in sediments collected from the River Ravi. Dicofol was found for the very first time in the biological samples from Pakistan, and its concentration was measured as relatively high in eggs from heronry from the River Chenab. Residual concentrations measured in eggs were below the levels that could affect egret populations. Biomagnification of the total OCPs through the food chain was evident in three breeding heronries. The concentration of DDE measured in eggs of the cattle egret suggests the need for monitoring this contaminant in other bird species at different trophic levels.     

Organochlorine Pesticides and PCBs in Wild Boars from Calabria (Italy)

At present, there are no specific studies on the evaluation of environmental toxicological risks in Calabria (south of Italy) and on the presence of contaminants in the fauna of this region. The aim of the present research was to investigate the levels of contamination by OC pesticides and PCBs in some organs and tissues of wild boars (utilized as 'biological indicator') from various areas of Calabria. Quantitative determinations of organochlorines were carried out using GC-ECD and confirmed with GC-MS in 154 samples from wild boars (heart, liver, lung, kidney, muscle tissue and spleen) during the hunting season from 2000 to 2002. The results indicate the low residual levels of DDE in 8 samples and DDT in 4 samples and PCBs residues (Aroclor 1232) below the detection limits were found. Therefore, these results show that the Calabria region is not at contamination risk from organochlorines and moreover is free from health problems for the consumer of boar meat.     

Mercury in wild mushrooms and underlying soil substrate from the great lakes land in Poland

Fifteen species of wild mushrooms and underlying soil samples collected in a virgin landscape of Augustowska Forest in northeastern Poland in 1997-98 were analyzed for total mercury to evaluate the status of contamination and usefulness of higher mushrooms as possible bioindicators of mercury pollution. Among the 15 species analyzed, Pinewood King Bolete, Scaly Tooth and King Bolete showed relatively high bioconcentration factors (BCF: dry-weight normalized concentrations of mercury in mushrooms relative to concentrations in soil) for mercury, which varied between 69 and 110. These three species were also characterized by great concentrations of total mercury in caps (between 2,000 +/- 800 and 2,300 +/- 1,100 ng g-1 dry wt) and stalks (between 850 +/- 390 and 1,000 +/- 500 ng g-1 dry wt.). Species such as Red-hot Milk Cap, Poison Pax and Common Chantherelle had mercury BCFs of less than 1, while Gipsy Bolete, Orange Birch Bolete, Brown Scaber Stalk, Variegated Bolete, Sandy Knight-cap and Yellow-cracking Bolete were weak or moderate mercury accumulators with BCFs between 1 and 40. Concentrations of mercury in mushrooms were greater than the tolerance limits suggested for mercury in plant foods.     

Volatile organic compounds (VOCs) emitted from materials collected from buildings affected by microorganisms

In this study mould damaged materials, including carpet, concrete, gypsum board, insulation, plastic, sand and wood, from 20 different buildings with moisture problems were collected. To study emissions from these materials both conventional methods for sampling, such as collection on Tenax TA, were used as well as complementary methods for sampling a wider spectrum of compounds, such as more volatile VOCs, amines and aldehydes. Analysis was carried out using gas chromatography and high-performance liquid chromatography. Mass spectrometry was used for identification of compounds. Alcohols and ketones were almost exclusively emitted from the materials after they had been wet for a week. Acids were also emitted in large quantities from wet gypsum board and plastic. No primary or secondary amines could be identified, but two tertiary amines, trimethylamine and triethylamine, were emitted from sand contaminated by Bacillus. The most common moulds found were Penicillium and Aspergillus. A multivariate method (partial least squares, PLS) was used to investigate the emission patterns from the materials. Materials with bacterial growth had a different VOC profile to those with only mould growth.     

Copper, zinc, and cadmium body concentrations in Hexaplex trunculus collected from the Tunisian coast

Copper, Zn, and Cd were analyzed in the soft tissues of the edible gastropod Hexaplex trunculus collected from seven localities along the Tunisian coast. The body concentration of these metals was significantly different depending on the gender and site. Copper and Zn varied, respectively, from 47.70 to 343.64 μg/g dry weight (dw) and 149.46 to 530.44 μg/g dw, being higher in males. Cadmium varied from 0.22 to 18.95 μg/g dw and was always higher in females. Overall, the highest concentrations of the three metals were all recorded in Gabès fishing harbor. Comparison of metal concentrations with the European standards compiled by the Food and Agriculture Organization showed that values exceeded standards in several localities. Imposex had been previously recorded in H. trunculus along the Tunisian coast; however, it does not seem to be related with Cu, Zn, and Cd pollution because the body concentration of these metals is not correlated with imposex degree. Nevertheless, interactive effects resulting from the combination of different pollutants and other stressors cannot be disregarded.     

Monitoring metal contamination levels and fecal pollution in clam (Tapes decussatus) collected from Izmir Bay (Turkey)

The aim of this study was to monitor the heavy metal contents and fecal pollution in Tapes decussatus (carpet shell clam) from Izmir Bay (Eastern Aegean). Bivalve mollusks were sampled on January, March, July, and October 2007 in the Izmir Bay. Izmir Bay is one of the great natural bays of the Mediterranean. Concentrations of heavy metals were determined in the clams from the different seasons. Fecal coliform densities were determined to evaluate the degree of water pollution and clams’ microbiological accumulation of the classical microbial pollution indicators. The concentration of heavy metals in T. decussatus from Izmir Bay were Hg 0.044–0.13; Cd 0.026–0.24; Pb 0.38–1.2; Cr 2.3–3.7; Cu 6.4–8.4; Zn 56.0–81.8, and Ni 8.1–9.6 μg/g (dry weight). The maximum values were generally obtained in July and March except Ni. This study found that the levels of heavy metals except Cr in T. decussatus were below Spanish and European Communities legislations for shellfish as food. Microbial pollution indicators (heterotrophic bacteria and fecal coliform) were measured in T. decussatus. Maximum heterotrophic bacteria and fecal coliforms were recorded in the winter while the lowest were detected in summer.     

水磨河流域有机氯农药污染调查及防治对策

在对水磨河流域实地勘察的基础上,用GC-ECD定量测定了乌鲁木齐市水磨河底泥中有机氯农药HCHs和DDTs的含量,以此来了解水磨河有机氯农药污染情况。结果表明,样品中HCHs和DDTs含量分别为0.107~111.690和0.476~66.512ng/g,其检出率为100%,说明该流域已经普遍受到有机氯农药污染。针对目前的污染现状提出相应的治理措施,以减轻水磨河有机氯农药的污染。     

Shell disturbances and butyltins burden in commercial bivalves collected from the Bizerta lagoon (northern Tunisia)

Shell disturbances and soft tissues butyltin burden were investigated in commercial bivalves Lithophaga lithophaga, Mytilus galloprovincialis, Solen marginatus and Crassostrea gigas from the Bizerta lagoon. Shell disturbances were found in all bivalves, being scarce in S. marginatus. In the internal valve of L. lithophaga, burrowing annelids and sipunculids living inside galleries were observed, while in the external valve, brown-blackish or white stains were found. In M. galloprovincialis, a yellowish mass located at the shell anterior side was found fixed firmly to the pearly layer by a hard brownish structure covering some annelid elliptic eggs. In the internal shell layer of some specimens collected in April, embryos belonging to tubiculous annelids at various developmental stages were observed. In C. gigas, shell thickening was revealed in some specimens corresponding to white doughy deposits at the internal valve and between shell layers. In S. marginatus, only one specimen showing a cavity at the posterior site was found. Total butyltin concentrations in the studied bivalves varied between 30 and 245?ng/g dry weight with tributyltin (TBT) being the predominant compound. The highest concentration was recorded in L. lithophaga collected from the Bizerta Bay and the lowest concentration in S. marginatus from Maghraoua. This study provided baseline data that could serve for long-term monitoring of TBT pollution in Tunisia, since legislation to reduce the use of TBT-based antifouling paints has not been introduced yet.     

Metal and metallothionein concentrations in Paracentrotus lividus from Amvrakikos gulf (Ionian Sea-Greece)

Concentrations of Cd, Cu, Cr, Ni, Zn, Fe and metallothioneins (MTs) were measured in the gonads of Paracentrotus lividus from Amvrakikos gulf (Ionian Sea, Greece). Three natural populations were selected; two of them, growing inside the gulf (Agios Thomas and Koronisia), presented higher density and smaller body size than the population living in a coastal area just outside the gulf (Mytikas). Metal and MT levels were not elevated, with the exception of Zn, showing high values related to the reproduction stage of the sea urchins. Significant geographical variations were recorded in the concentrations of Cu, Zn, Cd, Cr and MTs. The highest mean and maximum values of Cu, Zn and MTs were recorded in Agios Thomas while Cd and Cr were higher in Mytikas population. Copper, Zn, Fe and MT concentrations were negatively correlated to the sea urchin body size, while a positive concentration-size relationship was observed for Cd. Although all studied populations grow in a low metal level marine environment, urchins with smaller body size living in a food limited marine environment showed higher gonadosomatic index, metal concentrations and MT levels in their gonads (Agios Thomas and Koronisia) than larger specimens growing in a food unlimited area (Mytikas).     

Organochlorine pesticide residues in sediments from coastal environment of Cantabria (northern Spain) and evaluation of the Atlantic Ocean

This paper documents levels of organochlorine pesticides (OCs) in coastal surface sediments from selected reference sites on the northern Atlantic Spanish coast. One hundred eight samples covering three estuaries in the Cantabrian Coast were sampled in 2006 and analyzed in the finer fraction (<63 ??m) for 19 OCs by gas chromatography with electron capture detector after confirmation by mass spectrometry. Detected organochlorine pesticides were endosulfan ??, endosulfan ??, endosulfan sulfate, hexachlorobenzene (HCB), aldrin, dieldrin, methoxychlor, 1,1-dichloro-2,2-bis(4-chlorophenyl)ethylene (4,4??-DDE) and 1,1-dichloro-2,2-bis(4-chlorophenyl)ethane (4,4??-DDD). Total OCs concentrations ranged from 1.8 ng g^???1 dry weight (dw) to 3.9 ng g^???1 dw, showing a uniform distribution along the studied area, and being consistent with recorded levels in the literature for coastal sediments in other reference sites with low levels of pollution by OCs along the Atlantic Ocean. Endosulfan, 4,4??-DDD, HCB, aldrin, and dieldrin seemed to be ubiquitous as the legacy of past uses and deposition. OCs concentrations were significantly correlated to organic matter content and particle size distribution. No adverse biological effects derived from these pollutants are expected to take place as it can be concluded from the comparison with the existent marine sediment quality guidelines.     

Organochlorine pesticides and polychlorinated biphenyl residues in reared and wild Dicentrarchus labrax from the Mediterranean Sea (Sicily, Italy)

The aim of this research was to compare the levels of organochlorine pesticides and PCBs in samples of Dicentrarchus labrax living in the Straits of Messina with samples cultivated in cages in the Mediterranean Sea. Muscles and liver tissues sampled over the months, within the same year, were analyzed. The quantitative determination of the organochlorine compounds was performed by GC-ECD and confirmed with GC–MS. The results showed that the concentrations of DDTs in muscles and livers as such of reared sea bass were in the range 0.2–1.3 μg/kg and 9.6 –48.4 μg/kg, respectively. In wild fish the concentrations of DDTs were very much lower: 0.1 μg/kg in muscles, 5.1–9.0 μg/kg in livers. Total PCBs levels were higher in cultivated sea bass than in wild fish; the concentration ranges were 5.3–59.7 μg/kg and 74.4–267.4 μg/kg in muscle and liver of reared samples, respectively, and 1.1–1.5 μg/kg and 63.2–109.4 μg/kg in muscle and liver of wild samples, respectively.     

Spatial and Temporal Trends in Organochlorine Contamination and Bill Deformities in Double-Crested Cormorants (Phalacrocorax Auritus) from the Canadian Great Lakes

The levels of organochlorine contaminants (OCs) in the eggs of double-crested cormorants (Phalacrocorax auritus) from the Canadian Great Lakes, Lake Nipigon and Lake-of-the-Woods were monitored between 1970 and 1995. PCBs and p,p-DDE were present at the highest concentrations. Significant declines in OC concentrations on the Great Lakes were observed over this period for Lake Ontario, Lake Superior, Georgian Bay and North Channel but not Lake Erie where levels remained relatively stable. In the early 1970s, the greatest OC levels were generally observed in cormorant eggs from nesting sites in Georgian Bay and North Channel of Lake Huron. Between 1984 and 1995 mirex and PCB levels were consistently highest in samples from Lakes Ontario and Erie, respectively. Similar levels of PCDDs and PCDFs were observed from all regions of the Canadian Great Lakes in 1989. In general, OC levels in cormorant eggs between 1984-95 were ranked as follows: Lake Erie>Lake Ontario Lake Superior>Lake Huron. In 1995, eggshell thickness in Canadian Great Lakes cormorants, ranged from 0.423 to 0.440 mm and was on average only 2.3% thinner than pre-DDT era values. Between 1988 and 1996, 31 cormorant chicks with bill defects were observed at 16 different colonies (21% of all colonies surveyed) in Lakes Ontario and Superior, Georgian Bay and North Channel, and the main body of Lake Huron. No bill deformities were observed at reference sites in northwestern Ontario (Lake Nipigon and Lake-of-the-Woods). For the period 1988-96, the prevalence of bill defects in cormorant chicks (0.0 to 2.8/10,000 chicks) did not differ significantly (P > 0.05) among most regions in the Canadian Great Lakes. Georgian Bay was the only region to show a significant decrease in the prevalence of bill defects between the periods 1979-87 and 1988-95.     

Organochlorine pesticides and polychlorinated biphenyls in common buzzard (<Emphasis Type="Italic">Buteo buteo</Emphasis>) from Sicily (Italy)

In the present study, we investigated the concentrations and distribution of organochlorine pesticides (OCs) and polychlorinated biphenyls (PCBs) in intestine, liver, and muscle samples of 11 common buzzards (Buteo buteo) from Sicily used as bioindicator for monitoring pollution in environment. All samples of common buzzards were collected at the “Recovery Center of Wild Fauna” of Palermo, through the Zooprophilactic Institute. Quantitative determination of OCs and PCBs in the samples examined has been carried out using HRGC-ECD and GC-MS. The results obtained show the presence of concentrations of ∑DDT and ∑PCB in almost all samples. Regarding ∑DDT (4,4^′-DDE, 2,4^′-DDD, 4,4^′-DDD, 2,4^′-DDT, and 4,4^′-DDT), the highest concentrations were found in intestine (0.411 ± 0.050 μg/g) followed by muscle (0.130 ± 0.017 μg/g) and liver samples (0.109 ± 0.014 μg/g). As regards the ∑PCB congeners (PCB-28, PCB-52, PCB-95, PCB-99, PCB-101, PCB-110, PCB-138, PCB-146, PCB-149, PCB-151, PCB-153, PCB-170, PCB-177, PCB-180, PCB-183, and PCB-187), the highest concentrations were found in intestine (1.686 ± 0.144 μg/g) followed by liver (1.064 ± 0.162 μg/g) and muscle samples (0.797 ± 0.078 μg/g). Our data deserve particular attention not only for their significance but especially because they were recorded in Sicily, a region with a very low risk of environmental pollution due to the shortage of industries.     

Concentrations of perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) in vacuum cleaner dust collected in Japanese homes

Perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) are shown to be globally distributed, environmentally persistent and bioaccumulative. Although there is evidence that these compounds exist in the serum of non-occupationally exposed humans, the pathways leading to the presence of PFOS and PFOA are not well characterized. The concentrations of PFOS and PFOA in the vacuum cleaner dust collected in Japanese homes were measured. The compounds were detected in all the dust samples and the ranges were 11-2500 ng g(-1) for PFOS and 69-3700 ng g(-1) for PFOA. It was ascertained that PFOS and PFOA were present in the dust in homes, and that the absorption of the dust could be one of the exposure pathways of the PFOS and PFOA to humans. With regard to risk management, it is important to consider the usage of PFOS and PFOA in the indoor environment in order to avoid further pollution.     

Genotoxicity of the sediments collected from Pearl River in China and their polycyclic aromatic hydrocarbons (PAHs) and heavy metals

The accelerated industrialization and urbanization in the last three decades around the Pearl River Delta within Guangdong Province in China have led to serious concerns about the impacts on the aquatic environment. In the present study, the genotoxicity of the sediments collected from the Pearl River was evaluated by micronucleus (MN) assay with Vicia faba root tip cells, and the 16 EPA priority polycyclic aromatic hydrocarbons (PAHs) and heavy metals (HMs, including Cr, Cu, As, Se, Cd, Hg, and Pb) in the sediments were determined respectively by GC-MS, inductively coupled plasma mass spectrometry, and inductively coupled plasma atomic emission spectrometry. The results showed that there were significant increases of MN frequencies observed in the sediment-exposed groups, compared with the negative group (P?相似文献   

Imposex and butyltin concentrations in Bolinus brandaris (Gastropoda: Muricidae) from the northern Tunisian coast

The imposex incidence and butyltin concentration i.e. tributyltin with its di- and mono-substituted metabolites were investigated in the muricid Bolinus brandaris sampled from two sites on the northern Tunisian coast (the Lagoon of Bizerta and the small Gulf of Tunis). Both populations had imposex, with stages of imposex development varying between VDS 1 and VDS 4.3. All imposex indices (imposex frequency (I %), female penis length, female vas deferens length, vas deferens sequence index, relative penis length index, and vas deferens length index) were significantly higher in snails from the Bizerta lagoon. Butyltins were detected in the whole tissues of both sexes from the two sites. TBT levels were higher in gastropods collected from the lagoon of Bizerta (12.65 ±1.48 ng Sn g^???1 dw in female and 15.21 ±1.13 ng Sn g^???1 dw in male) than in individuals from the Gulf of Tunis (10.71 ±1.26 ng Sn g^???1 dw in female and 11.65 ±1.63 ng Sn g^???1 dw in male), corroborating the data of imposex analysis. These results confirmed that B. brandaris could be used as a bioindicator species of butyltin pollution in the studied areas. In addition, this study provided baseline data that could serve for long-term monitoring of TBT pollution in Tunisia, since legislation to reduce the use of TBT-based antifouling paints has not been introduced yet.     

Metal concentrations and metallothionein levels in Mytilus galloprovincialis from Elefsis bay (Saronikos gulf, Greece)

Spatial and temporal variability of Cd, Cu, Cr, Ni, Zn, Fe and Mn and metallothionein (MT) concentrations were determined in mussels Mytilus galloprovincialis from Elefsis bay (Saronikos gulf, Greece). Higher concentrations of both metal concentrations and MTs were recorded in mussels inhabiting industrial locations (steelworks and shipyard), indicating a markedly higher metal bioavailability. However at these sites, located at the eastern part of the bay, mussel metal concentrations were not always correlated with both seawater metal concentrations and MTs possibly due to different time scales of integration of the metal sources into mussels and/or the participation of other metal regulatory mechanisms except MT induction. The pattern of the temporal variation of mussel metal concentrations and the MT levels was similar among stations with higher values during the winter–spring season and lower during the summer–autumn period. The inverse relationship of flesh condition index with mussel metal concentrations was attributed to the influence of mussel annual reproductive cycle.     

Trace metal concentrations in suspended particles, sediments and clams (Ruditapes philippinarum) from Jiaozhou Bay of China

Suspended particulate matter (SPM), sediments and clams were collected at three sites in Jiaozhou Bay to assess the magnitude of trace metal pollution in the area. Metal concentrations in SPM (Cu: 40.11–203; Zn: 118–447; Pb: 50.1–132; Cd: 0.55–4.39; Cr: 147.6–288; Mn: 762−1670 μg/g), sediments (Cu: 17.64–34.26; Zn: 80.79–110; Pb: 24.57–49.59; Cd: 0.099–0.324; Cr: 41.6–88.1; Mn: 343−520μg/g) and bivalves (Cu: 6.41–19.76; Zn: 35.5–85.5; Pb: 0.31–1.01; Cd: 0.51–0.67; Mn: 27.45−67.6 μg/g) are comparable to those reported for other moderately polluted world environments. SPM showed a less clear pattern. Metal concentrations in sediments displayed a clear geographical trend with values increasing with proximity to major urban centers. The clams (on dry weight) showed a complex pattern due to the variability introduced by age-related factors. Cd showed an apparent reverse industrial trend with higher concentrations in clams collected at distant stations. Zn, Pb and Mn showed no clear geographical pattern, whereas Cu increased in the clams collected in the most industrialized area. In addition, the bioaccumulation factors (BAF) were calculated. The result indicated that the studied Ruditapes philippinarum in Jiaozhou Bay possessed different bioaccumulation capacities for Cd, Zn, Cu, Pb and Mn, and Cd, Zn had a relatively high assimilation of those metals from sediment particles. A significant relationship with clam age was observed for Zn (positive) and Cu (negative) suggesting different physiological requirements for both metals with age. Trace metal concentrations measured in the tissue of the investigated clam were in the range considered safe by the WHO for human use.     

Trace metal concentrations in suspended particles, sediments and clams (Ruditapes philippinarum) from Jiaozhou Bay of China

Suspended particulate matter (SPM), sediments and clams were collected at three sites in Jiaozhou Bay to assess the magnitude of trace metal pollution in the area. Metal concentrations in SPM (Cu: 40.11-203; Zn: 118-447; Pb: 50.1-132; Cd: 0.55-4.39; Cr: 147.6-288; Mn: 762-1670 microg/g), sediments (Cu: 17.64-34.26; Zn: 80.79-110; Pb: 24.57-49.59; Cd: 0.099-0.324; Cr: 41.6-88.1; Mn: 343-520 microg/g) and bivalves (Cu: 6.41-19.76; Zn: 35.5-85.5; Pb: 0.31-1.01; Cd: 0.51-0.67; Mn: 27.45-67.6 microg/g) are comparable to those reported for other moderately polluted world environments. SPM showed a less clear pattern. Metal concentrations in sediments displayed a clear geographical trend with values increasing with proximity to major urban centers. The clams (on dry weight) showed a complex pattern due to the variability introduced by age-related factors. Cd showed an apparent reverse industrial trend with higher concentrations in clams collected at distant stations. Zn, Pb and Mn showed no clear geographical pattern, whereas Cu increased in the clams collected in the most industrialized area. In addition, the bioaccumulation factors (BAF) were calculated. The result indicated that the studied Ruditapes philippinarum in Jiaozhou Bay possessed different bioaccumulation capacities for Cd, Zn, Cu, Pb and Mn, and Cd, Zn had a relatively high assimilation of those metals from sediment particles. A significant relationship with clam age was observed for Zn (positive) and Cu (negative) suggesting different physiological requirements for both metals with age. Trace metal concentrations measured in the tissue of the investigated clam were in the range considered safe by the WHO for human use.