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Evaluation of the effects of cryopreservation of isolated erythrocytes and leukocytes of largemouth bass by flow cytometry (1995)

Fisher, S. K. ; Lingenfelser, J. T. ; Jagoe, C. H. ; [et al.]

Oxford, UK : Blackwell Publishing Ltd

Journal of fish biology 46 (1995), S. 0

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ISSN: 1095-8649

Source: Blackwell Publishing Journal Backfiles 1879-2005

Topics: Biology

Notes: We examined the effects of separation and freezing on fish leukocyte and erythrocyte morphology by light microscopy and on DNA content as measured by flow cytometry (FCM). Leukocytes and erythrocytes of largemouth bass Micropterus salmoides were isolated by density gradient centrifugation of whole blood, and frozen in liquid nitrogen in a buffer containing DMSO as a cryopreservative. The coefficient of variation (CV) of the G0/G1 peak of the cells was used to assess variation in nuclear DNA content within cell populations before and after separation and freezing treatments. In erythrocytes, the CV did not change significantly (P〉0.05) when nuclei were isolated and stained without freezing or when erythrocytes were frozen prior to nuclear isolation and staining. In leukocytes, freezing and thawing prior to isolation and staining of nuclei significantly increased the CV (P〈0.05), and produced hyperdiploid shoulders of the G0/G1 peak. However, the CV of leukocyte nuclei that were isolated and stained prior to freezing and the CV of non-frozen leukocyte nuclei did not differ (P〉0.05). Microscopy showed that the freezing protocol had little effect on erythrocyte morphology, but caused irregular swelling in leukocytes. Freezing intact leukocytes also significantly (p〈0.05) altered the apparent distribution of cells among the phases of the cell cycle as measured by FCM. The distributions of leukocyte nuclei that were isolated and stained prior to freezing were not different to non-frozen leukocytes. DNA measurements of nucleated blood cells are widely used in physiological, genetic and toxicological studies. Our results suggest that whole blood and erythrocytes for use in such studies can be frozen whole using a simple protocol, but leukocyte nuclei must be isolated and stained before freezing to avoid serious artifacts.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1111/j.1095-8649.1995.tb05983.x

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Tissue Mercury Concentrations in Alligators (Alligator mississippiensis) from the Florida Everglades and the Savannah River Site, South Carolina (1997)

Yanochko, G. M. ; Jagoe, C. H. ; Brisbin Jr., I. L.

Springer

Archives of environmental contamination and toxicology 32 (1997), S. 323-328

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ISSN: 1432-0703

Source: Springer Online Journal Archives 1860-2000

Topics: Energy, Environment Protection, Nuclear Power Engineering , Medicine

Notes: Abstract. Mercury pollution is a serious problem in some areas of the southeastern United States. Due to biomagnification, long-lived predators should have high Hg concentrations in affected areas. American alligators (Alligator mississippiensis) are important predators in many southwestern wetlands, but little information is available on Hg concentrations in this species. We collected tissues from alligators inhabiting two sites in the Everglades, Florida (n = 18) and a manmade reservoir in South Carolina (Par Pond; n = 44), all with documented histories of Hg contamination, and analyzed them for total Hg. Mean concentrations in kidney, liver, muscle, and dermal scutes of alligators from the Everglades (expressed as mg Hg/kg dry mass ± S.E.M.) were 36.42 ± 5.23, 41.09 ± 5.90, 5.57 ± 0.47, and 5.83 ± 1.04, respectively. Concentrations in liver, muscle, and scutes from Par Pond alligators were 17.73 ± 2.56, 4.08 ± 0.46, and 4.58 ± 0.63, respectively. Blood from Par Pond alligators contained 2.20 ± 0.38 mg Hg/kg wet mass. Mercury concentrations did not differ among sexes at any location. Tissue Hg levels did not differ significantly between Everglades locations, but were lower in Par Pond. In Everglades alligators, Hg concentrations in all tissues were positively correlated, as were tissue Hg and total length. Only total length and scute Hg were correlated in Par Pond alligators. Regression revealed a significant relationship between muscle and scute Hg concentrations in Everglades alligators, but not Par Pond alligators. Alligators living in polluted areas can accumulate substantial concentrations of Hg, but relationships among Hg concentrations in specific tissues may vary with location or age and size of the animals sampled.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/s002449900192

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Dietary Exposure to Mercury in Nestling Wood Storks (Mycteria americana) in Georgia (1998)

Gariboldi, J. C. ; Jagoe, C. H. ; Bryan, Jr., A. L.

Springer

Archives of environmental contamination and toxicology 34 (1998), S. 398-405

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ISSN: 1432-0703

Source: Springer Online Journal Archives 1860-2000

Topics: Energy, Environment Protection, Nuclear Power Engineering , Medicine

Notes: Abstract. The United States breeding population of the wood stork (Mycteria americana) was listed as endangered in 1984; the U.S. Fish and Wildlife Service has listed the determination of potential contaminant hazards as a task important to the recovery of the species. Storks nest in colonies and adult storks forage over wide areas and return with prey items to feed to nestlings. The most recently consumed prey items are usually regurgitated by nestlings when they are handled. In 1995, over 200 food items from four wood stork colonies were collected while banding nestlings between 20 and 40 days old, and total mercury determined in individual food items. Two colonies were located on the Atlantic coast and two in inland areas. Overall dietary mercury concentration for each colony was calculated based on the biomass of each type of food item within the diet. Freshwater prey items had higher mercury concentrations than saltwater items most frequently chosen by storks. Daily Hg doses were estimated for a range of nestling sizes and daily food consumption rates. Risk factors were calculated based on two lowest observable adverse effect concentrations (LOAECs) from previous avian mercury ingestion studies. Based on one LOAEC, all four colonies in this study were at risk of sublethal effects due to mercury in the diet of nestling wood storks. More extensive studies of sublethal effects in avian species from mercury contamination are necessary for more accurate risk assessment.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/s002449900336

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Perch mercury content is related to acidity and color of 26 Russian lakes (1995)

Haines, T. A. ; Komov, V. T. ; Matey, V. E. ; [et al.]

Springer

Water, air & soil pollution 85 (1995), S. 823-828

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ISSN: 1573-2932

Keywords: fish ; mercury ; lake chemistry ; biomagnification

Source: Springer Online Journal Archives 1860-2000

Topics: Energy, Environment Protection, Nuclear Power Engineering

Notes: Abstract We determined mercury in fish (perch Perca fluviatilis) from 26 Russian lakes in three regions over four years. The lakes ranged in size from 2 to 395,000 ha, in pH from 4.5 to 10.0, and in color from 3 to 190 hazen. Sixteen lakes were drainage lakes, with permanent outlets, and 10 were seepage lakes, with no permanent inlets or outlets. The lakes were generally located in forested regions with little or no human habitation in the watershed. The three regions were geologically distinct: Precambrian Shield granitic bedrock covered with thin soil; Triassic bedrock covered with thick glacial tills; and Triassic bedrock covered with thin sediments. At each lake water samples were collected and analyzed for pH, add neutralizing capacity (ANC), major cations, and anions. Dissolved mercury species were estimated with a thermodynamic equilibrium model (MINTEQA2). Mercury content of dorsal muscle varied from 0.04 to 1.0 μg/g wet weight, and was linearly related to calculated HgCH3Cl (r20.68, p〈0.001). Lake HgCH3Cl, in turn, was related to lake pH (r2=0.86, p〈0.001). Stepwise multiple regression selected lake HgCH3Cl and color as the factors most highly related to fish mercury content, with the model accounting for 75% of the variation.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF00476931

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