Dolphin Kidney Research Articles

Subcellular Cadmium, Lead and Mercury Compartmentalization in Guiana Dolphins (Sotalia guianensis) From Southeastern Brazil

Marine mammals are considered excellent ocean health sentinels and are ubiquitously exposed to chemical contaminants worldwide. The Guiana dolphin (Sotalia guianensis) is a near-threatened dolphin species from Brazil with unknown population size data. This indicates the need for assessments regarding deleterious effects that may arise from exposure to chemical contamination, especially metals. After entry in the organism, these compounds are subject to internal subcellular compartmentalization, which in turn alters their bioavailability. However, almost no assessments regarding subcellular metal contents in marine mammals are available. In this context, metal compartmentalization was determined in three subcellular fractions for three toxic elements, Cd, Hg and Pb, by inductively coupled plasma mass spectrometry (ICP-MS) in Guiana dolphin kidney and liver samples from Southeastern Brazil. Differential metal-detoxification mechanisms were observed for the three elements, where metallothionein (MT) detoxification was postulated for only for Pb, while Cd and Hg were poorly associated to MT, and mostly present in the insoluble fraction, indicating low bioavailability. This is the first report on subcellular metal compartmentalization in Guiana dolphins and indicates that critical biochemical detoxification data is obtained through subcellular fraction analyses in marine mammals. This indicates an emerging study field for this type of assessment, which may, in turn, aid in conservation efforts.

Crystallization of Hierarchical Ammonium Urate: Insight into the Formation of Cetacean Renal Stones

Among a wide variety of pathological crystalline materials in nephrolithiasis, uric acid and its salt form are the most abundant organic constituents. Even though rarely found in humans, ammonium urate (NH4HU) is the most prevalent kidney stone reported in managed bottlenose dolphins. In this study, we investigate the physicochemical properties of NH4HU crystals associated with dolphin kidney stones, which exhibit distinct hierarchical structures. We present a method of crystallizing NH4HU without common impurities, which allowed for the first measurement of NH4HU crystal solubility over a broad range of temperatures. Parametric evaluations of NH4HU crystallization are reported, wherein we assess the effects of supersaturation and solution alkalinity on crystal phase behavior, the kinetics of urate crystallization, and the size and morphology of synthetic NH4HU crystals. In vitro bulk crystallization assays from aqueous growth solutions containing supersaturated ionic species (NH4+ and urate1–) were perfo...

Determination of cadmium and lead in cetacean Dolphinidae tissue from the coast of Bahia state in Brazil by GFAAS

In the present study, cadmium and lead in the muscle, lung, liver and kidney of dolphins ( Sotalia guianensis and Stenella clymene) of the Bahia coast in the northwest of Brazil were determined by graphite furnace atomic absorption spectrometry. Samples were digested using a diluted oxidant mixture (HNO 3 + H 2O 2) with a microwave heating program performed in five steps. The optimized temperatures and chemical modifier for the pyrolysis and atomization were 700 °C, 1400 °C and Pd plus Mg for Cd, and 900 °C, 1800 °C and NH 4H 2PO 4 for Pb, respectively. Characteristic masses and limits of detections ( n = 20, 3 σ) for Cd and Pb were 1.6 and 9.0 pg and 0.82 ng g − 1 and 0.50 ng g − 1 , respectively. Repeatability ranged from 0.87 to 8.22% for Cd and 4.31 to 8.09% for Pb. The found concentrations presented no statistical differences at the 95% confidence level when compared with the ICP OES methods. Addition and recovery tests were also performed and the results ranged between 87 and 112% for both elements. Samples of cetacean Dolphinidae ( S. guianensis and S. clymene) were analyzed, and the higher concentrations ranged from 0.09 to 46.2 µg g − 1 for Cd and 0.04 to 0.47 µg g − 1 for Pb in liver, and from 0.133 to 277 µg g − 1 for Cd in the kidney.

Perfluorooctane sulfonate (PFOS), perfluorooctanoic acid (PFOA) and their salts Scientific Opinion of the Panel on Contaminants in the Food chain.

Perfluorooctane sulfonate (PFOS), perfluorooctanoic acid (PFOA) and their salts Scientific Opinion of the Panel on Contaminants in the Food chain.

Elemental composition of liver and kidney tissues of rough-toothed dolphins (Steno bredanensis).

On December 14, 1997, 62 rough-toothed dolphins (Steno bredanensis) stranded on Cape San Blas, on the Florida coast of the Gulf of Mexico. Approximately 30 animals died either on the beach or in rehabilitation facilities. Two were successfully rehabilitated and released. Liver, kidney, blubber, and muscle tissues were collected from 15 animals that died on the beach. Portions of the liver and kidney from each dolphin were analyzed using instrumental neutron activation analysis and inductively coupled plasma mass spectrometry to determine mass fractions of 37 elements. Levels of several electrolytes (Na, Cl, K, Br, Rb, I, Cs) and of the essential trace elements Fe, Cu, and Zn in both tissues were similar to those found in other Odontoceti. Mass fractions of Ca ranged from 60 mg/kg to 1,200 mg/kg (wet mass basis), indicating significant inhomogeneity in the kidney tissues of several animals. Necropsy reports noted that the kidneys of many of these animals contained fibrous nodules. The measured Ca inhomogeneity may be due to mineralization of the fibrous kidney tissue. Hepatic levels of Hg and Se were at the high end of the ranges generally found in livers of other Odontoceti and were slightly higher in animals with fibrous kidneys than in the others. Mass fractions of Se, Ag, and Hg in liver tissues increased with the size and age of the animals indicating accumulation of these elements in the liver with age. Results also indicate that Se and Hg accumulate in rough-toothed dolphin kidney. Accumulation of these elements with age has been reported commonly for marine mammals and other species.

Inhibition of gap junctional intercellular communication by perfluorinated compounds in rat liver and dolphin kidney epithelial cell lines in vitro and Sprague-Dawley rats in vivo.

Gap junctional intercellular communication (GJIC) is the major pathway of intercellular signal transduction, and is thus important for normal cell growth and function. Recent studies have revealed a global distribution of some perfluorinated organic compounds, especially perfluorooctane sulfonic acid (PFOS) in the environment. Because other perfluoroalkanes had been shown to inhibit GJIC, the effects of PFOS and related sulfonated fluorochemicals on GJIC were studied using a rat liver epithelial cell line (WB-F344) and a dolphin kidney epithelial cell line (CDK). In vivo effects on GJIC were studied in Sprague-Dawley rats orally exposed to PFOS for 3 days or 3 weeks. Effects on GJIC were measured using the scrape loading dye technique. PFOS, perfluorooctane sulfonamide (PFOSA), and perfluorohexane sulfonic acid (PFHA) were found to inhibit GJIC in a dose-dependent fashion, and this inhibition occurred rapidly and was reversible. Perfluorobutane sulfonic acid (PFBS) showed no significant effects on GJIC within the concentration range tested. A structure activity relationship was established among all 4 tested compounds, indicating that the inhibitory effect was determined by the length of fluorinated tail and not by the nature of the functional group. The results of the studies of the 2 cell lines and the in vivo exposure were comparable, suggesting that the inhibitory effects of the selected perfluorinated compounds on GJIC were neither species- nor tissue-specific and can occur both in vitro and in vivo.

Effects of benzo(a)pyrenf and tetrachlorodibenzo(p) dioxin on fetal dolphin kidney cells: Inhibition of proliferation and initiation of DNA damage

Dolphin kidney cells (CDK) were exposed in vitro to benzo(a)pyrene (BaP) in the presence or absence of 2,3,7,8-tetrachlorodibenzo(p)dioxin (TCDD), a cytochrome P450-inducing agent, and/or α-naphthoflavone (α NF), an inhibitor of cytochrome P450 induction. BaP inhibited mitosis in CDK cells in a dose-dependent manner. TCDD, while inhibiting cell proliferation, did not show a strict dose-dependent mode of action. BaP inhibition of mitosis was decreased by αNF, which also decreased the inhibitory effects of TCDD on CDK proliferation. BaP treatment initiated both 3H-thymidine incorporation and the increased alkali lability of DNA functions of the initiation of excision repair. Cells pre-treated with TCDD and then exposed to BaP exhibited increased BaP-DNA adduct levels and increased DNA excision repair. These data indicate that dolphin cells metabolized BaP in vitro as a function of cytochrome P450-associated activities, that BaP metabolites covalently bound to cellular DNA and initiated excision repair. Inhibition of the cytochrome P450-mediated metabolism of BaP decreased the BaP-associated inhibition of mitosis in dolphin cells.

CHARACTERIZATION OF A BOTTLENOSE DOLPHIN (TURSIOPS TRUNCATUS) KIDNEY EPITHELIAL CELL LINE

Abstract: An epithelial cell line, Carvan dolphin kidney (CDK), isolated from a prematurely born female bottlenose dolphin, Tursiops truncatus, exhibited growth characteristics not previously reported for cetacean cells in culture. CDK cells were cytokeratin positive and demonstrated a maximum doubling time of 1.31 days, with plating and colony forming efficiencies approaching 100% for the early population doublings. Despite an unusually efficient colony‐forming ability and rapid growth, these cells were neither transformed nor immortal, displaying normal contact inhibition, anchorage dependence, and the requirement for high concentrations of fetal bovine serum in the growth medium. CDK cells exhibited age‐dependent changes in growth rate, colony‐forming efficiency, and cytoplasmic profile, and showed a finite lifespan of about 50 population doublings and a stable 2N = 44 karyotype which correlates with previously reported cytogenetic analyses. Velocity sedimentation analysis showed that CDK cells contained nuclear aryl hydrocarbon receptor (Ah), indicating their potential to be induced for cytochrome P450. These data suggest that CDK cells may have utility as an in vitro toxicological model for evaluating hydrocarbon contaminant effects on Tursiops truncatus, a protected marine mammal.

Susceptibility of a line of dolphin kidney cell culture to several herpesviruses.

A cell line was established from cell cultures of kidney cortex of a pantropical spotted dolphin, Stenella attenuate. The replication of 6 strains of herpesviruses was studied in the cells. Five strains of them, herpes simplex virus type I and type II, equine rhinopneumonitis virus, infectious bovine rhinotracheitis virus and Aujeszky's disease virus, were grown fairly well in showing clear cytopathic effects and plaques under agar overlay medium.

Organochlorine chemical and heavy metal contaminants in white-beaked dolphins (Lagenorhynchus albirostris) and pilot whales (Globicephala melaena) from the coast of Newfoundland, Canada.

Polychlorinated biphenyls (∑PCB), DDT isomers (∑DDT), chlordanes (∑CHLOR), toxaphene, chlorobenzenes and hexachlorocyclohexane isomers and seven elements (As, Cd, Cu, Hg, Pb, Se, Zn) were determined in tissues of 41 stranded pilot whales (Globicephala melaena) and 27 ice-entrapped white-beaked dolphins (Lagenorhynchus albirostris) from Newfoundland, Canada. Cadmium, Hg, As and Se were significantly higher in pilot whale kidney and liver than in dolphin tissues. Mercury in liver and blubber, and Cd in kidney, of pilot whales were positively correlated with age. Cadmium levels in both species were much higher than reported for other cetaceans from Canadian east coast waters. Lead concentrations in dolphin kidney and mucles were 5 times higher than in pilot whales. Levels of ∑PCB (sum of 49 congeners) in blubber samples ranged from 31 to 61 mg/kg (lipid wt) in female and male dolphins, respectively, and from 5.6 to 12 mg/kg in female and male pilot whales. ∑DDT and toxaphene were present at similar levels to ∑PCB while mean ∑CHLOR (sum of seven components) ranged from 1.6 to 17 mg/kg in blubber of pilot whales and dolphins, respectively. The presence of high levels of toxaphene in the blubber of both species was unexpected and may be due to increased use of this pesticide during the 1970's. Higher levels of all organochlorine chemicals in blubber, as well as Pb in kidney and muscle, of dolphins than in pilot whales may reflect greater exposure to contaminants because of overwintering and feeding in Gulf of St. Lawrence waters.

Acidic glycolipids from dolphin kidney.

The composition and contents of acidic glycolipids in the kidney of a striped dolphin (Stenella coeruleoalba, the order Cetacea, whales) were determined. The following eight acidic glycolipids were isolated and characterized: SM4s (124.2 nmol/g tissue), SM3 (8.7), GM3 (NeuAc) (12.3), GM3 (NeuGc) (31.6), GD3 (NeuAc-NeuAc) (14.7), GD3 (NeuAc-NeuGc) (II3 alpha(NeuAc alpha 2-8NeuGc)-LacCer) (9.8), GD3 (NeuGc-NeuAc) (II3 alpha(NeuGc alpha 2-8NeuAc)-LacCer) (5.3), and GD3 (NeuGc-NeuGc) (15.8). The assignment of the four types of GD3 was further confirmed as described below. Evidence indicating 2-8 linkages of the disialosyl residues of GD3 was obtained on methylation analysis of sialic acid. GD3 (NeuAc-NeuAc) and GD3 (NeuGc-NeuAc) were degraded to GM3 (NeuAc), and GD3 (NeuAc-NeuGc) and GD3 (NeuGc-NeuGc) yielded GM3 (NeuGc) on mild acid hydrolysis. Fragment ions characteristic of the carbohydrate and lipophilic moieties of the permethylated GD3 were observed in direct inlet-electron impact-mass spectra. The presence of these four types of GD3 in a tissue has not been reported previously. GD3 contained non-hydroxy (69-84%) and hydroxy fatty acids (16-31%) with 16-24 carbons. The long chain base of all GD3, except GD3 (NeuGc-NeuAc) (not analyzed), consisted of 4-sphingenine (d18:1) and 4-hydroxysphinganine (t18:0) in almost equal amounts. The total amount of renal lipid-bound acidic groups (sulfate and sialic acid) of the dolphin (190 mumol/animal) is considerably higher than that of a terrestrial mammal (88 mumol/animal) with a body weight comparable to that of the dolphin. This deviation suggests that the amount of renal acidic amphiphiles required to maintain the osmotic balance of body fluids in marine mammals might be higher than that in terrestrial ones.

Growth potential of sheep and sea mammal cells transformed by SV40 early region DNA.

AbstractNormal fibroblasts of bearded seal, northern fur seal, sea lion, domestic sheep, and normal kidney cells of spotted dolphin and Pacific common dolphin have been transformed by fragments of SV40 DNA containing the entire early region. The transformed, T-antigen-positive cell strains grew to higher saturation densities and, except for the transformed dolphin kidney cell strains, were able, albeit poorly, to form colonies in soft agar medium. The transformed northern fur seal, spotted dolphin and Pacific common dolphin cell strains have remained diploid, the transformed sheep cells have become hypodiploid due to Robertsonian translocations, and the transformed bearded seal and sea lion cells had become hypotetraploid. Although all the transformed cells have acquired increased divisional potential and prolonged lifespan in cell culture, only the hypotetraploid cell strains of bearded seal and sea lion have evolved into continuous cell strains.

Replication of measles virus in dolphin kidney cells.

Replication of measles virus in dolphin kidney cells.

Water and Electrolyte Content of Dolphin Kidney and Extraction of Pressor Substance (Renin)

1. The values for the water content and electrolyte concentrations of a normal dolphin kidney were as follows: Total water, 821.0 g; chloride, 65.3 mM; sodium, 84.2 mM; potassium, 56.8 mM; calcium, 1.53 mM; and magnesium, 6.3 mM per kg of fat-free tissue.