Concentrations of Polychlorinated Biphenyls (PCB’s), Chlorinated Pesticides, and Heavy Metals and Other Elements in Tissues of Belugas, Delphinapterus leucas, from Cook Inlet

Tissues from Cook Inlet beluga whales, Delphinapterus leucas, that were collected as part of the Alaska Marine Mammal Tissue Archival Project were analyzed for polychlorinated biphenyls (PCB’s), chlorinated pesticides, and heavy metals and other elements. Concentrations of total PCB’s (ΣPCB’s), total DDT (ΣDDT), chlordane compounds, hexachlorobenzene (HCB), dieldrin, mirex, toxaphene, and hexachlorocyclohexane (HCH) measured in Cook Inlet beluga blubber were compared with those reported for belugas from two Arctic Alaska locations (Point Hope and Point Lay), Greenland, Arctic Canada, and the highly contaminated stock from the St. Lawrence estuary in eastern Canada. The Arctic and Cook Inlet belugas had much lower concentrations (ΣPCB’s and ΣDDT were an order of magnitude lower) than those found in animals from the St. Lawrence estuary. The Cook Inlet belugas had the lowest concentrations of all (ΣPCB’s aver-aged 1.49 ± 0.70 and 0.79 ± 0.56 mg/kg wet mass, and ΣDDT averaged 1.35 ± 0.73 and 0.59 ± 0.45 mg/kg in males and females, respectively). Concentrations in the blubber of the Cook Inlet males were significantly lower than those found in the males of the Arctic Alaska belugas (ΣPCB’s and ΣDDT were about half). The lower levels in the Cook Inlet animals might be due to differences in contaminant sources, food web differences, or different age distributions among the animals sampled. Cook Inlet males had higher mean and median concentrations than did females, a result attributable to the transfer of these compounds from mother to calf during pregnancy and during lactation. Liver concentrations of cadmium and mercury were lower in the Cook Inlet belugas (most cadmium values were <1 mg/kg and mercury values were 0.704–11.42 mg/kg wet mass), but copper levels were significantly higher in the Cook Inlet animals (3.97–123.8 mg/kg wet mass) than in Arctic Alaska animals and similar to those reported for belugas from Hudson Bay. Although total mercury levels were the lowest in the Cook Inlet population, methylmercury concentrations were similar among all three groups of the Alaska animals examined (0.34–2.11 mg/kg wet mass). As has been reported for the Point Hope and Point Lay belugas, hepatic concentrations of silver were re

DNA extraction from formalin-fixed Franciscana tissues

The present paper reports the extraction of DNA from formalin-fixed Pontoporia blainvillei tissues. Following the Vachot and Monerot (1996) protocol, fragmented DNA (300-700bp) was extracted from more than 95% of liver and muscle samples. DNA yield in liver samples was significantly higher than in muscle samples (4.574 ± 1.169mg DNA/mg versus 0.808 ± 0.297mg DNA/mg). Similar results were obtained from nine other species of cetaceans and five species of pinnipeds. It is of special interest to have a method that allows the utilisation of museum specimens not originally preserved for genetic studies, which may include rarely available, declining or extinct species. SPANISH: El presente trabajo reporta la extracción de ADN a partir de tejidos formolizados de Pontoporia blainvillei. Siguiendo el protocolo de Vachot y Monerot (1996) se pudo extraer ADN degradado (300-700pb) en más del 95% de las muestras de hígado y músculo analizadas. El rendimiento en ADN fue significativamente mayor en muestras de hígado que en muestras de músculo (4.574 ± 1.169mg DNA/mg tejido húmedo versus 0.808 ± 0.297mg DNA/mg tejido húmedo). Resultados similares se obtuvieron en otras nueve especies de Cetáceos y cinco de Pinnípedos. Resulta de gran interés contar con un método que permita la utilización de especímenes depositados en museos y que no hayan sido originalmente colectados para estudios genéticos, incluyendo especies de difícil obtención, en franca declinación o extintas.

Preliminary report on bottlenose dolphin (Tursiops truncatus) uterine samples for parity analysis

There have been numerous studies on various mammalian species regarding vascular changes in uterine arteries elucidating the effects of parity. In equids, vascular changes of uterine arteries have been demonstrated to occur in uniparous and multiparous mares. The severity of these arteriole changes suggests a link to previous pregnancies. Differences in the number or range of pregnancies can be ascertained through microscopic evaluation of elastin deposition in the arterioles, perivascular fibrosis, and stromal cellularity. There has been little, if any, work performed on parity in the bottlenose dolphin (Tursiops truncatus). The objective of this preliminary study was to determine the feasibility of detecting similar vascular changes in the endometrium of known-aged female bottlenose dolphins to assess parity. Archived formalin fixed samples of uterus were obtained from nine bottlenose dolphins with known age and parity. Four slides were made from each sample and individually stained with four different techniques. From our small sample pool, it appears that uteri from nulliparous animals do not develop perivascular fibrosis. Parous uteri developed perivascular fibrosis and arteriolar elastosis. These changes agree with our expectations that some degeneration (elastosis) and compensation (fibrosis) occurs as a result of uterine expansion of pregnancy. The assessment of this technique for use in bottlenose dolphins would provide an important tool in the determination of the reproductive success of dolphin populations, identify individuals who are sexually mature but nulliparous, which could indicate reproductive dysfunction or increased calving intervals, and increase our knowledge on the role contaminants play in reproductive dysfunction.

Ascorbic acid turnover in the ocular tissues of some fishes

The ascorbic acid turnover from the ocular tissues of 11 species of fishes from a culture pond and the river Godavari (Andhra Pradesh, India) has been studied. The free ascorbic acid and ascorbigen contents were more in the case of bottom or deep dwelling fishes and the least in the case of surface living forms, depending upon the light penetration in the area that each species inhabits. The enzymic utilization and ascorbic acid-macromolecule complex varied among the fishes possibly depending upon individual energy requirements and not upon light intensity. No size-related or sex-related variation was observed. No variation was observed between riverine and pond-reared fishes of the same species.

Determination of ubiquinone and tocopherol in heart, liver and muscle tissues of fresh water fish, Rohu (Labeo rohita, Hamilton)

The liver, heart and muscle tissues of Rohu, (Labeo rohita, Hamilton) were examined for their ubiquinone (UQ) and tocopherol contents. These three tissues contained respectively 11.60, 3.94 and 0.19 mg of ubiquinone and 10.16, 5.32 and 3.58 mg of tocopherol per 100 g. The 4% (V/V) of diethyl ether (EE) in light petroleum ether (PE) fractions of all three tissues on paper chromatographic separation gave spots having the same Rf value as standard ubiquinone-50 (UQ 10). Both the 4% and 6% (V/V) diethyl ether in light petroleum ether fraction of liver, heart and muscle tissues gave a single spot with the same Rf value as α – tocopherol.

Palatability of tissues in Holothuria leucospilota (Brandt) from central west coast of India

The palatability of various organs (body wall, cuvierian gland, viscera, longitudinal muscle bands and gonads) of sea cucumber Holothuria leucospilota (Brandt) was studied by feeding experiments, performed on a freshwater fish Sarotherodon mossambicus and a marine fish Therapon jarbua. The result shows that the food pellets of the body wall were less toxic and more palatable than the gonads, viscera and cuvierian gland (p<0.001).

Distribution of 5'-nucleotidase activity in the tissues of a freshwater teleost, Ophicephalus striatus Bloch

Homogenates of tissues have been analysed for the presence of 5'-nucleotidase activity. Sexes are not treated individually since no significant differences were observed. All organs showed marked activity, the highest being in the cardiac tissues, then the kidney, spleen, brain, and low values in the liver and muscles. Comparison with data from other fish studied shows a marked difference between O. striatus and O. punctatus.

Survival and changes in the fine structure of selected tissues of Penaeus monodon Fabricius juveniles fed various carbohydrates

Penaeus monodon juveniles were reared on semipurified diets containing various carbohydrates (maltose, sucrose, dextrin, molasses, cassava starch, corn starch or sago palm starch). Significant differences were observed between the type as well as the level of carbohydrate in the diet on the survival of the juveniles. Results indicate that there does not seem to be any correlation between survival and the complexity of the carbohydrates.

Determination of bioaccumulation and microscopic survey of LAS pollutant effect on the liver, kidney and gill tissues of Rutilus frisii kutum in Caspian sea

Linear alkylbenzene sulfonate (LAS) are widely used in detergent industry. Due to contaminants entering the water, and the effects of their accumulation in fish, LAS, has a great importance in environmental pollution. In the present study, accumulation of LAS and its histological effects on gill tissue, liver and kidney of Caspian kutum (Rutilus frisii kutum) were studied. Caspian kutum is the most important and most valuable teleosts of the Caspian Sea. Due to releasing Caspian Kutum in rivers and Anzali Lagoon and unlimited entry of wastewater to the aquatic ecosystem, research on the impact of LAS on Caspian kutum is important. In the present study, fish exposed to sublethal concentrations of LAS (0.58, 1.16 and 2.32 mg/l) for 192 hours. Control treatments with three replicates at 0, 24, 48, 72, 96 and 192 hours were done. For assessments of the histological effects of LAS, tissue sections prepared and by using Hematoxylin - Eosin were stained, then the prepared sections, examined by light microscopy. For determination of the bio accumulation of LAS, the soxhlet extraction and solid phase extraction was performed to determine the amount of LAS using HPLC with fluorescence detector. According to results average of bioconcentration factor and LAS concentrations in fish had reached stable levels after approximately 72 h and thus represented steady state BCF values in this species. The value of steady-state bio-concentration factor of total LAS was 33.96 L.Kg- 1 and for each of the homologous C10-n-LAS, C11-n-LAS, C12-n-LAS and C13-n- LAS were 3.84, 6.15, 8.58 and 15.57 L.Kg-1 respectively. According to the results obtained in gills exposed to LAS, histopathological alteration include hypertrophy, lifting of lamella epithelium, edema, clubbing of lamellae hyperplasia, lamellar fusion and aneurysm were seen. In liver tissue exposed to three concentrations of LAS, congestion and dilation of sinusoids, irregular-shaped nuclei and degeneration in the hepatocyte, vacuolar degeneration and necrosis were observed. In kidney exposed to three concentrations of LAS, reduction of the interstitial haematopoietic tissue, degeneration in the epithelial cells of renal tubule, tubular degeneration, necrosis, shrinkage and luminal occlusion were observed. According to the results the most alteration due to exposure to LAS was seen in the gill tissue. None of the control samples showed histological effects of LAS.

The study of biological characteristics and heavy metal determination of Cd, Pb, Fe, V, Ni in the muscle, liver and gonad tissues of kingfish Scomberomorus commerson in the coastal waters of the Hormozagan Province (Persian Gulf)

Biological studies and heavy metal (Ni, v, Fe, Pb,Cd) determination in liver, gonad and muscle of Scomberomorus commerson were carried out from Oct 2006 to Sept 2007 in Hormozgan coastal waters. 599 Samples were gutted for reproduction and nutrition studies, fork length and weight were measured to nearest cm and g respectively. Meanwhile 40 samples were also investigated for heavy metal studies. All specimens were collected from two major landing sites (Bandar Lengeh & Bandar Abbas). Minimums & maximum fork length & weight were 29, 128 cm & 235 and 15350 g respectively.Isometric growth was shown according to our study and b was estimated 2.9 (overall), 2.91(male) & 2.89 (female). The average relative gut length was 0.52± 0.007 and it was determined that S. commerson is a carnivorous. More than 99 percent of gut content was different teleost fishes. Gastro somatic index had two peaks in Nov & Jan (before spawning) and with a decreased trend in July, the spawning period. Occurrence of empty stomach was estimated % 65.77. Maximum amount of condition factor was in Dec. Spawning season was started from June. The average of Absolute & relative fecundity (to weight unit) was 1217149±179315 and 178.2±15.58 respectively. Lm50% was estimated 75 cm for females. Sex ratio was 0.97: 1 (male: female). Chi- Square test showed no significant difference (p>0.05). Maximum amount of hepatosomatic index was estimated in March.Metal concentrations were determined using either Flame Atomic Absorption Spectroscopy (for Fe) or Graphite Furnace Atomic Absorption Spectroscopy (for Pb,Cd,Ni and V). The mean concentration (μg/g dry weight)of Pb,Cd,Ni,V and Fe in the liver were 0.0309, 0.0268, 0.0672, 0.0077, 2.5159 in the gonad 0.0440 ,0.0295, 0.1096, 0.0000, 1.4449 and in the muscle 0.0244, 0.0324, 0.0656, 0.0128, 1.6138 respectively. The maximum metal concentrations were below the maximum permissible limits for human consumption recommended by the USEPA, WHO and the UK. The results of Kendall's Tau-b correlation coefficient were as follows: The Liver tissue: There were significant positive linear relationships between accumulation of V, Fe, and Pb with Fork length, Pb and Fe with weight, GSI with Pb, Cd, V and 109 Fe, and a negative linear relationships between HSI with accumulation of V and Fe, Fork length, weight and GSI. The Gonad tissue: There were significant positive linear relationships between GSI with accumulation of Pb, Cd, Fe, Fork length and weight, a negative linear relationship between HSI with Fork length, weight and GSI. The Muscle tissue: There were significant positive linear relationships between accumulations of V, Fe with Fork length and weight factors and as well as GSI with Cd, V, Fe, Pb, Fork length and weight,a negative linear relationship between HSI with Fork length, weight, Cd, Fe and GSI. The results of Mann-Whitney U tests (P≤0.05) show that there were significant differences between summer and autumn from heavy metal contents in the studied tissues point of view. The only exceptions were for Ni in the liver, gonads and muscle and as well as there were significant differences between male and female from heavy metal contents in the studied tissues. The only exceptions were for Pb in muscle, Ni in liver, gonad and muscle, V in muscle, and Cd and Fe in gonads.