Organochlorine exposure and health effects in stranded Steller sea lions (Eumetopias jubatus) and Pacific harbor seals (Phoca vitulina richardii) from Oregon and southern Washington coasts

Organochlorine (OC) pesticides and polychlorinated biphenyls (PCBs) have been detected in a variety of marine mammal species at levels associated with adverse health effects. Little is known about OC levels and impacts on health in pinnipeds with different life histories. We determined the health and levels of 18 OC pesticides and 16 PCB congeners in blubber samples from 20 Steller sea lions and 39 Pacific harbor seals stranded from Oregon and Southern Washington. The most commonly detected OC at the highest concentration was p,p′- dichlorodiphenyldichloroethylene (DDE). PCBs were detected in all samples as well. Hypothesis testing indicated that diseased Steller sea lions (males and females combined) had higher contaminant concentrations than healthy Steller sea lions, and diseased Pacific harbor seals had higher concentrations of total OCs than healthy animals. Differences were also noted between diseased and healthy animals when looking at individual sexes of each species. Diseased Steller sea lions had higher mean contaminant levels than diseased harbor seals and healthy Steller sea lions had higher mean contaminant concentrations than healthy Pacific harbor seals. These results show that species differences exist in both contaminant loads and sensitivity to contaminants, which may be due to differences in life histories and physiology.     

Laboratory study of heavy metal phytoremediation by three wetland macrophytes

Detention ponds and constructed wetlands have proven to be effective in reducing peak stormwater runoff volume and flow, and recent interest has extended to utilizing them to improve stormwater runoff quality. A review of stormwater runoff studies indicated that lead, zinc, copper, cadmium, phosphorus, and chloride are contaminants of primary concern. In laboratory settings, the uptake of contaminants by three wetland plant species, Glyceria grandis, Scirpus validus, and Spartina pectinata, was examined and removal rates from nutrient solutions inflow and nonflow reactors were measured. The removal rates varied by plant species and target contaminant, and no one species was the best accumulator of all six contaminants. Belowground tissues of all three species accumulated higher concentrations of the four heavy metals and aboveground tissues accumulated higher concentrations of phosphorus and chloride. Plants grown in flow reactors showed significantly higher accumulation rates than those grown in nonflow reactors. Also, plants grown hydroponically accumulated higher concentrations of the six target contaminants than those grown in sand reactors. However, those grown in sand had a much greater increase of biomass and removed a greater mass of the six target contaminants. Removal rates measured in these experiments can be used to design detention ponds to maximize stormwater remediation.     

Effects of yo-yo diet, caloric restriction, and olestra on tissue distribution of hexachlorobenzene

Chlorinated hydrocarbons are lipophilic, toxic, and persistent in the environment and animal tissues. They enter the body in food and are stored in adipose tissue. Loss of body fat through caloric restriction mobilizes stored lipophilic xenobiotics and results in distribution to other tissues. We have studied the reversibility of this process in mice that followed a regimen of body weight cycling. Weight gain was followed by weight loss, a second gain, and a second loss ("yo-yo diet regimen"). We measured the distribution of orally gavaged [14C]hexachlorobenzene, which is sparingly metabolized. We found that weight cycling has different effects in different organs. Continued weight loss resulted in a threefold increase of 14C amount and concentration in the brain. After weight regain, 14C in the brain decreased but then increased again after a second weight loss. Weight loss resulted in an increase in the concentration of 14C in adipose tissue without changing the total amount in that tissue. Weight loss and regain resulted in an increase of 14C in the liver, which reflected an increase of fat in the liver. The regimen of weight gain and loss was repeated in mice gavaged with [14C]hexachlorobenzene, with one group receiving the nonabsorbable fat olestra in the diet. Combined dietary olestra and caloric restriction caused a 30-fold increase in the rate of excretion of 14C relative to an ad libitum diet or a reduced caloric diet alone. Distribution of 14C into the brain resulting from the restricted diet was reduced by 50% by dietary olestra.     

The influence of breeding colony and sex on mercury,selenium and lead levels and carbon and nitrogen stable isotope signatures in summer and winter feathers of <Emphasis Type="Italic">Calonectris</Emphasis> shearwaters

Contamination in marine foodwebs is nowadays of great environmental concern owing to the increasing levels of pollution in marine ecosystems from different anthropogenic sources. Seabirds can be used as indicators of regional contaminant patterns across large temporal and spatial scales. We analysed Hg, Se and Pb levels as well as stable isotope ratios of C (¹³C/¹²C, δ¹³C) and N (¹⁵N/¹⁴N, δ¹⁵N) in breeding- and winter-season feathers on males and females of two related shearwater species, providing information on spatiotemporal patterns of contaminants as well as the influence of the trophic ecology of these seabirds on contaminant levels. During the breeding season, Se and Pb concentrations were highest at the Cape Verde archipelago, showing no differences among the other colonies or between the sexes. However, Hg levels varied among colonies, being highest in the Mediterranean, probably resulting from the larger emissions and fallout of this pollutant in Europe. Feathers grown during breeding also showed sexual differences in Hg concentrations and δ¹³C. Differences in Hg concentration between sexes are mainly due to egg-laying decontamination in females. In contrast, differences in Hg among colonies are probably related to differences in trophic ecology, as indicated by δ¹³C and δ¹⁵N measurements. Contaminant concentrations in winter-grown feathers did not show any relationship with stable isotope values but were affected by contaminant loads associated with the breeding season. These findings suggest that the interpretation of contaminant levels of migratory species from feathers moulted out of the breeding season should be made with caution because those values could reflect exposures to contaminants acquired during the breeding season. We conclude that factors other than feeding ecology may play an important role in the interpretation of contaminant levels and their annual dynamics at several spatial scales. Consideration of the relevant temporal context provided by isotopic signatures and contaminant concentrations is important in deciphering contaminant information based on various tissues. Xavier Ruiz: deceased 27 April 2008.     

Biological monitoring of heavy-metal contamination in coastal waters off Casey Station, Windmill Islands, East Antarctica

Heavy-metal concentrations were determined in tissues of different species of benthic invertebrates collected in the Casey region (Australian Antarctic Territory) where an old waste-disposal tip site is a source of contamination. The species studied included the bivalve Laternula elliptica, starfish Notasterias armata, heart urchins Abatus nimrodi and A. ingens and gammaridean amphipod Paramoera walkeri. The specimens were collected at both reference and contaminated locations where lead was the priority element and copper was the next most important in terms of increased concentrations. The strong association between a gradient of contamination and concentrations in all species tested indicates that they are reflecting well the environmental changes, and that they appear as appropriate biological indicators of heavy-metal contamination. Aspects of the biology of species with different functional roles in the marine ecosystem are discussed in relation to their suitability for wider use in Antarctic monitoring programmes. For example, in terms of heavy-metal bioaccumulation, the bivalve appears as the most sensitive species to detect contamination; the starfish provides information on the transfer of metals through the food web while the heart urchin and gammarid give indications of the spatial and temporal patterns of the environmental contamination. The information gathered about processes of contaminant uptake and partitioning among different tissues and species could be used in later studies to investigate the behaviour and the source of contaminants.     

The use of rotifers for the maintenance of monoalgal mass cultures of Spirulina

Zooplankton was successfully used for the biological control of unicellular algal contaminants in Spirulina mass cultures even under conditions adverse to the growth of Spirulina (maximal winter daily temperature of approximately 10 degrees C and very low bicarbonate concentration). Brachionus plicatilis (Rotifera) was the most successful species of zooplankton used. The interrelationships between Spirulina, green unicellular contaminant, and B. plicatilis were studied under various conditions. Two species of unicellular contaminant were used; Monoraphidium minutum was isolated from local cultures and Chlorella vulgaris, obtained from contaminated Spirulina cultures in Israel. The rotifer B. plicatilis successfully controlled the population size of both contaminants whether they were introduced in a single addition or as a daily dose. The biological control of the unicellular contaminants allows Spirulina to be cultured in a medium low in bicarbonate, thereby reducing the cost of the medium and increasing the quantity of CO(2) that may be freely absorbed from the atmosphere at the optimal pH for Spirulina cultivation.     

Distribution of chlorpromazine and imipramine in adipose and other tissues of rats

Adipose tissue kinetics of chlorpromazine and imipramine, two drugs which are more lipophilic than thiopental, were studied in the rat. After single i.v. doses, the time-course of drug distribution was followed in adipose and various other tissues, until their concentrations in adipose tissues declined. Under these conditions the two drugs behaved almost identically. Among the tissues analyzed, the lowest concentrations were found in adipose tissue, with the exception of plasma. At its maximum concentration after about 30 minutes, total adipose tissue contained only 3 % of the dose of administered drugs. Adipose/plasma and adipose/lung concentration ratios were 2–5 and 0.05, respectively.After maximum tolerated oral doses of imipramine for 3 weeks, similar steady state concentration ratios (plasma:adipose:brain:lung 1:3:12:96) were observed. In adipose tissue the imipramine/desmethylimipramine ratio was about 1, and the desmethylimipramine steady state levels did not increase with time. Literature data indicate that many basic lipophilic drugs are not stored in adipose tissue. This is now clearly shown for chlorpromazine and imipramine, even under extreme, subchronic conditions in the case of imipramine.     

Accumulation of organochlorines and mercury in flounder — an approach to pollution assessments

Levels of PCBs, HCB and Hg were determined in the muscle tissue of flounder (Platichthys flesus L.), sampled at two sites of the highly industrialized Elbe estuary which is characterized by a decreasing gradient of contaminant levels in water and sediments towards the mouth of the river. The contaminant levels were compared to those found in muscle tissue of flounders caught in an unpolluted reference area, the Eider estuary. Juvenile flounders (1–2 cm total length) caught during their migration from the southern North Sea coast into the Elbe estuary in early summer showed a significantly higher level of contamination at both Elbe stations than those from the reference area. In the following spring, the flounders, having grown to a length of 12–14 cm, reflected the gradient of contaminants measured in water and sediment. However, during summer and late autumn a drastic increase in average contaminant concentration was observed in the muscle tissue of flounders taken from the less polluted estuarine site. It became evident that severe oxygen depletion in the heavily polluted area near Hamburg had provoked the migration of highly contaminated fish downstream to the less polluted sampling area; thus intermingling with the less contaminated flounder population of this region had occurred. The results obtained indicate that careful analysis of environmentally controlled behavioural patterns in the species used for monitoring purposes as well as an appropriate statistical treatment of the data gained from chemical analysis are essential for assessing the contaminant concentrations in organisms in relation to the pollution load of their habitats.     

Prevalence of skin ulceration in cod (<Emphasis Type="Italic">Gadus morhua callarias</Emphasis> L.) under anthropogenic contamination in the southeastern part of the Baltic Sea

Data on the occurrence of skin ulceration cases and concentrations of anthropogenic contaminants (PCBs; DDT; PAN; and heavy metals Hg, Pb, As, and Cd) in Baltic cod in the Russian exclusive economic zone (EEZ) of the southeastern part of the Baltic Sea are presented and an attempt is made to analyze the relationship between the prevalence of disease and contaminant concentrations. In 2005–2009 the prevalence of skin ulcers was 0.7%. The share of skin ulceration in fish was maximal in 3- and 6-year-old cod (1.0 and 1.1%, correspondingly) and in fish of the size group 39–45 cm (0.9%). Since 2007, a reduction of skin ulceration cases has been recorded in the Russian EEZ of the southern Baltic Sea. In 2005–2009, the concentration of contaminants in cod tissues did not exceed the permissible levels accepted in the Russian Federation. The relationship between the concentration of PCBs in skin and the prevalence of ulceration cases requires further studies.     

Aggregative oviposition behaviour of Helicoverpa spp. (Lepidoptera: Noctuidae) in contaminated chickpea crops

Abstract Aggregative egg distribution by Helicoverpa spp. within experimental field plots of chickpea containing various contaminant plant species was documented at different stages of plant growth. More eggs were laid on contaminant faba and wheat plants than on the surrounding chickpea. The level of egg aggregation on faba and wheat plants increased with general crop growth, becoming evident before a significant disparity in growth rates between these species and the surrounding chickpea became apparent. In chickpea plots contaminated with canola, aggregative oviposition activity on the contaminant plants was observed in the early seedling stage whilst a clear height differential between the two species was evident. Total Helicoverpa egg density on contaminant plants was not correlated with height above chickpea for any of the contaminant species. It is proposed that the aggregative oviposition behaviour of Helicoverpa spp. in contaminated chickpea results from differential apparency of the contaminant plants to ovipositing moths. Differential apparency may result from several factors including growth rate and morphological differences between chickpea and the contaminants. Physiological changes in chickpea, including growth stage-related changes in acid production by the foliage, may also contribute to the greater apparency of contaminant plants in chickpea plots. The implications of differential host plant apparency for insect pest management are discussed.     

Hepatic antioxidant responses related to levels of PCBs and metals in chicks of three Arctic seabird species

The efficiency of antioxidant defenses and relationship with body burden of metal and organic contaminants has not been previously investigated in arctic seabirds, neither in chicks nor in adults. The objective of this study was to compare such defenses in chicks from three species, Black-legged kittiwake (Rissa tridactyla), Northern fulmar (Fulmarus glacialis), and Herring gull (Larus argentatus), and the relationship with tissue concentrations of essential metals such as selenium and iron and halogenated organic compounds, represented by polychlorinated biphenyl (PCB). The results showed significant species-specific differences in the antioxidant responses which also corresponded with metal and PCB levels in different ways. The capability to neutralize hydroxyl radicals (TOSC-HO?) and the activities of catalase and Se-dependent glutathione peroxidases (GPX) clearly increased in species with the higher levels of metals and PCBs, while the opposite trend was observed for Se-independent GPX, TOSC against peroxyl radicals (ROO?) and peroxynitrite (ONOOH). Less clear relationships were obtained for glutathione levels, GSH/GSSG ratio, glutathione reductase and superoxide dismutase. The results showed differences in antioxidant efficiency between the species, and some of these defenses exhibited dose-response-like relationships with measured levels of selenium, iron and ΣPCBs. PCBs, selenium and iron levels were positively related to the responses of antioxidants with potential to reduce HO?/H?O? (Se-dependent GPX, CAT and TOSC against HO?). However, direct causal relationships between antioxidant responses and contaminant concentrations could not be shown on individual level. Varying levels of metals and contaminants due to different diet and age were probably the main explanations for the species differences in antioxidant defense.     

Antibody biodistribution coefficients

Tissue vs. plasma concentration profiles have been generated from a physiologically-based pharmacokinetic model of monoclonal antibody (mAb). Based on the profiles, we hypothesized that a linear relationship between the plasma and tissue concentrations of non-binding mAbs could exist; and that the relationship may be generally constant irrespective of the absolute mAb concentration, time, and animal species being analyzed. The hypothesis was verified for various tissues in mice, rat, monkey, and human using mAb or antibody-drug conjugate tissue distribution data collected from diverse literature. The relationship between the plasma and various tissue concentrations was mathematically characterized using the antibody biodistribution coefficient (ABC). Estimated ABC values suggest that typically the concentration of mAb in lung is 14.9%, heart 10.2%, kidney 13.7%, muscle 3.97%, skin 15.7%, small intestine 5.22%, large intestine 5.03%, spleen 12.8%, liver 12.1%, bone 7.27%, stomach 4.98%, lymph node 8.46%, adipose 4.78%, brain 0.351%, pancreas 6.4%, testes 5.88%, thyroid 67.5% and thymus is 6.62% of the plasma concentration. The validity of using the ABC to predict mAb concentrations in different tissues of mouse, rat, monkey, and human species was evaluated by generating validation data sets, which demonstrated that predicted concentrations were within 2-fold of the observed concentrations. The use of ABC to infer tissue concentrations of mAbs and related molecules provides a valuable tool for investigating preclinical or clinical disposition of these molecules. It can also help eliminate or optimize biodistribution studies, and interpret efficacy or toxicity of the drug in a particular tissue.     

Procedure for isolation of gangliosides in high yield and purity: simultaneous isolation of neutral glycosphingolipids

While several methods for ganglioside extraction and isolation have been described, relatively little attention has been given to the effectiveness of separation from peptides, phospholipids, and various low-molecular-weight contaminants. A procedure is described for isolation of gangliosides in high purity and good yield from 1- to 400-mg samples (wet wt). A key step was mild acidification following homogenization, designed to dissociate gangliosides from lipophilic peptides which coextracted into organic solvents. This has proved particularly helpful for myelin and myelin-containing tissues (e.g., white matter, nerve) whose proteins have presented special problems in ganglioside isolation. In this study isolation was effected by consecutive chromatographies on Sephadex LH-20, DEAE-Sephadex, and silica gel following the initial acidification. The method applied to bovine white matter gave tissue concentrations (calculated from yields and radiolabeled tracer recoveries) that were similar to those obtained with three previously described procedures; however, peptide contaminants were an order of magnitude lower. Removal of low-molecular-weight contaminants, including nucleotide sugars, was virtually complete. In addition to ganglioside isolation the method can be used to obtain neutral glycosphingolipids as well. It is believed to have broad applicability to a diversity of tissues.     

High-performance liquid chromatographic assay of propofol in human and rat plasma and fourteen rat tissues using electrochemical detection

This paper describes a sensitive HPLC-electrochemical detection analytical method for determining the concentration of the intravenous anesthetic, propofol, in human or rat plasma or serum and a variety of rat tissues. Internal standard and drug are extracted from serum or plasma and other tissues with pentane. 2,6-tert.-Butylmethylphenol is used as internal standard. It includes a novel steam distillation procedure for separating the highly lipophilic propofol from skin and fat. The plasma/serum assay has a precision of 1–4% (C.V.) in the range 10 ng/ml to 1 μg/ml and permits the assay of 5 ng/ml from 0.1 ml of plasma/serum. The tissue procedure allows the estimation of 50 ng/g in 0.1 g of tissue for most of the major organs with less than 2% (C.V.) precision. This assay was used to measure propofol concentrations in plasma/serum and tissue samples in support of a project to develop a physiological pharmacokinetic model for propofol in the rat.     

Variation in tissue element concentration in mire plants over a range of sites

Elements in surface peat and ground water were estimated on seven mire sites in central and north Sweden. The variation in tissue clement concentration was studied in eleven plants occurring on the studied sites. The elements estimated were: N, P, K. Ca, Mg, Fe, Mn, Zn, Al, Cu, Mo, Na, S, B, and Si. The species studied were; Carex diandra, Carex limosa, Carex rostrata, Epilobium palustre, Menyanthes trifoliate, Pedicularis palustris, Rumex acetosa, Saxifraga hirculus, Scirpus hudsonianus, Stellaria crassifolia and Tofteldia pusilla . In surface peat over the range of sites, the variation in total concentrations of nitrogen and phosphorus was six and tenfold, respectively. The variation in total concentration of potassium was four-fold. Extractable concentrations of these three elements showed an even greater variation between sites. Over the range of sites, the variation in tissue element concentration was larger in roots than in above-ground parls. In above-ground parts, floral parts varied less in tissue element concentration than did stems and leaves. The largest variation in tissue element concentration was found in Carex rostrata and Menyanthes trifoliata . The smallest variation was found in Saxifraga hirculus . Tissues of the rich fen species Saxifraga hirculus and Stellaria crassifolia had the lowest concentrations of heavy metals. The correlation between substrate element concentration and tissue concentration differed both between tissues and species. Concentrations of N, P and K in tissues of the Carex species were highly correlated with the concentrations of these elements in the substrate, while N, P and K concentrations in tissues of the herbaceous species, especially Saxifraga hirculus . correlated less well.     

Whole-body to tissue concentration ratios for use in biota dose assessments for animals

Environmental monitoring programs often measure contaminant concentrations in animal tissues consumed by humans (e.g., muscle). By comparison, demonstration of the protection of biota from the potential effects of radionuclides involves a comparison of whole-body doses to radiological dose benchmarks. Consequently, methods for deriving whole-body concentration ratios based on tissue-specific data are required to make best use of the available information. This paper provides a series of look-up tables with whole-body:tissue-specific concentration ratios for non-human biota. Focus was placed on relatively broad animal categories (including molluscs, crustaceans, freshwater fishes, marine fishes, amphibians, reptiles, birds and mammals) and commonly measured tissues (specifically, bone, muscle, liver and kidney). Depending upon organism, whole-body to tissue concentration ratios were derived for between 12 and 47 elements. The whole-body to tissue concentration ratios can be used to estimate whole-body concentrations from tissue-specific measurements. However, we recommend that any given whole-body to tissue concentration ratio should not be used if the value falls between 0.75 and 1.5. Instead, a value of one should be assumed.     

Effect of dietary fat on metabolism and DNA adduct formation after acute oral exposure of F-344 rats to fluoranthene

Adverse health effects such as cancer and toxicity may be attributed to consumption of chemically contaminated food rich in fat. This leads to a larger intake and retention of lipophilic toxic chemicals in the body with an increase in risks to human health. The objective of this study was to characterize the effect of dietary fat on disposition and metabolism of fluoranthene (FLA), a polycyclic aromatic hydrocarbon compound. FLA was administered to F-344 rats in monounsaturated (peanut oil), polyunsaturated (corn oil) and saturated (coconut oil) fats at doses of 50 and 100 microg/kg via oral gavage. Blood, small intestine, liver, lung, testis, adipose tissue, urine and feces were collected at various time points' post-FLA exposure. Samples were analyzed by reverse-phase high-performance liquid chromatography for FLA parent compound and metabolites. DNA was isolated from the tissues and subjected to (32)P-post labeling to measure FLA-DNA adducts. The concentrations of unchanged FLA (FLA parent compound) and its metabolites showed an increase for the saturated fat treatment group compared with mono- and polyunsaturated fat groups. The FLA-DNA adduct concentrations were high in tissues of rats that received FLA through saturated fat. The toxicokinetic parameters, concentrations of FLA metabolites and FLA-DNA adduct showed a dose-dependent increase, and this increase was statistically significant (P<.05) for saturated fat. These findings clearly demonstrate that the high residence time of FLA parent compound in saturated fat allows extensive metabolism, contributing reactive metabolites of FLA that bind with DNA and causing marked damage in a long-term exposure scenario.     

Acid phosphatase activity in liver macrophage aggregates as a marker for pollution-induced immunomodulation of the non-specific immune response in fish

The activity of acid phosphatase in liver macrophage aggregates (MA-AP) of different fish species was used as a marker for a pollution-induced modulation of the digestive capacity of phagocytes, since functions of the non-specific immune response play a central role in the maintenance of animals' health. Based upon the investigation of more than 900 individual flounders (Platichthys flesus) and mullets (Liza aurata), natural variations, gender-specific differences and pollution-induced alterations in AP activity are demonstrated in this study. MA-AP activity was dependent on temperature and season but, nevertheless, distinctions between differently polluted areas were visible in all sampling campaigns with lowest MA-AP activity in fish from the polluted areas of the German Bight and the Israeli coast of the Mediterranean Sea. For organochlorine contaminants, as well as for mercury and copper, a significant correlation could be observed between residue concentrations in fish tissues and MA-AP activity. In all cases, except mercury which showed a positive correlation, AP activity was suppressed in animals with a high contaminant burden. MA-AP activity turned out to give reliable and consistent results for a quantification of immunomodulation in both fish species.     

CYTOCHROME P450 1A1 EXPRESSION IN CETACEAN INTEGUMENT: IMPLICATIONS FOR DETECTING CONTAMINANT EXPOSURE AND EFFECTS

Contaminant related health risks to marine mammals are typically inferred from the levels of contaminants measured in blubber. Such measurements alone are insufficient to indicate the likelihood of biological effects from contaminant exposure, especially for contaminants that do not bioaccumulate. Cytochrome P450 1A1 (CYP1A1) in mammals is induced by, and involved in, the metabolism of planar halogenated aromatic hydrocarbons and polycyclic aromatic hydrocarbons, chemicals of concern in aquatic systems. CYP1A induction is a molecular response to exposure to these inducers in many vertebrates. Using immunohistochemistry, we semiquantitatively measured CYP1A1 expression in integument (epidermis and blubber) collected by biopsy or at necropsy from 17 species of cetaceans. CYP1A1 expression was detected in all species and, in some cases, varied both within and between species. CYP1A1 expression in mysticetes was comparable to that in odontocetes. Assessing how the differences in contaminant burdens, life history parameters, and physiological condition between individuals, populations, or species affect CYP1A1 expression in cetacean integument is essential to the interpretation of this induction as a biomarker of exposure to and effects of contaminants. Detection of CYP1A1 expression in integument samples offers a relatively simple, non-lethal technique to study biological changes associated with contaminant exposure in cetaceans.     

In vivo lipolysis in adipose tissue from two anatomical locations measured by microdialysis

In this study, the difference in lipolytic response in inguinal subcutaneous and epididymal adipose tissues of male Sprague-Dawley rats was assessed in vivo by microdialysis. Probes were perfused with Ringer solution in which increasing concentrations of isoproterenol (10(-7) - 10(-4) mol/L) were added. Glycerol release, expressed as extracellular glycerol concentration, was used as lipolytic index. The effect of isoproterenol on local blood flow was investigated using the ethanol technique. No differences were found in the interstitial glycerol concentration between both adipose tissues under basal conditions. When isoproterenol was perfused, a dose-response increase in glycerol production was induced in both tissues. Interstitial glycerol concentration from epididymal adipose tissue was higher than that of inguinal subcutaneous depot at all isoproterenol concentrations. No vasodilatory effect of isoproterenol was found. These results suggest that epididymal adipose tissue is more responsive in vivo to beta-adrenergic lipolysis stimulation than is subcutaneous fat pad from the inguinal region.