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      Effects of Organochlorine Contaminants on Loggerhead Sea Turtle Immunity: Comparison of a Correlative Field Study and In Vitro Exposure Experiments

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          Abstract

          Several laboratory and field studies indicate that organochlorine contaminants (OCs), such as poly-chlorinated biphenyls (PCBs) and pesticides, modulate immune responses in rodents, wildlife, and humans. In the present study we examined the effects of OCs on immunity in free-ranging loggerhead sea turtles ( Caretta caretta). Mitogen-induced lymphocyte proliferation responses, lysozyme activity, and OC concentrations were measured from blood samples. Mitogens chosen in the lymphocyte proliferation assay were phytohemagglutinin (PHA) and concanavalin A (ConA) for T-lymphocyte stimulation, and lipopolysaccharide (LPS) and phorbol 12,13-dibutyrate (PDB) for B-lymphocyte stimulation. Lysozyme activity was significantly and negatively correlated with whole-blood concentrations of 4,4′-dichlorodiphenyldichloroethylene (4,4′-DDE) and the sum of chlordanes. Lymphocyte proliferation responses stimulated by PHA, LPS, and PDB were significantly and positively correlated with concentrations of the sum of PCBs measured in whole blood. LPS- and PDB-induced proliferation were also significantly and positively correlated with 4,4′-DDE blood concentrations. These correlative observations in free-ranging turtles suggest that current, chronic exposure to OCs may suppress innate immunity and enhance certain lymphocyte functions of loggerhead sea turtles. To further test this hypothesis, lymphocyte proliferation was measured after in vitro exposure of peripheral blood leukocytes from 16 turtles to Aroclor 1254 (0–13.5 μg/mL) or 4,4′-DDE (0–13.4 μg/mL). Both contaminants increased PHA- and PDB-induced proliferation at concentrations below those that affected cell viability. Moreover, the concentrations that enhanced PDB-induced proliferation in vitro were similar to concentrations measured in turtles with the highest proliferative responses. The similarities between the in vitro experiments and the correlative field study suggest that OC exposure modulates immunity in loggerhead turtles.

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          Most cited references38

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          The immediate effects of stress on hormones and plasma lysozyme in rainbow trout.

          The fight-or-flight response prepares an animal for coping with alarming situations and their potential consequences, which include injury. The possible involvement of innate components of immunity in the response has received little attention. We determined plasma concentrations of stress hormones and lysozyme activity before and after a 10 min handling stressor. Rainbow trout (Oncorhynchus mykiss) were anesthetized in their home tanks, bled, revived, and then stressed by being held in the air in a net for 30 s and placed in a shallow bucket of water for 10 min. Fish were then captured, concussed (in one of two experiments) and bled again. Control fish were also bled twice, but were kept anesthetized in their holding tanks between bleedings. Following the stressor, plasma cortisol, adrenaline and lysozyme activity were significantly increased. The experiment was repeated 4 months later with a similar outcome. While chronic stress is eventually immunosuppressive, acute stress/trauma may help enhance both cellular and humoral components of innate defenses at times of likely need.
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            Associations between Organochlorine Contaminant Concentrations and Clinical Health Parameters in Loggerhead Sea Turtles from North Carolina, USA

            Widespread and persistent organochlorine (OC) contaminants, such as polychlorinated biphenyls (PCBs) and pesticides, are known to have broad-ranging toxicities in wildlife. In this study we investigated, for the first time, their possible health effects on loggerhead sea turtles (Caretta caretta). Nonlethal fat biopsies and blood samples were collected from live turtles for OC contaminant analysis, and concentrations were compared with clinical health assessment data, including hematology, plasma chemistry, and body condition. Concentrations of total PCBs (∑PCBs), ∑DDTs, ∑chlordanes, dieldrin, and mirex were determined in 44 fat biopsies and 48 blood samples. Blood concentrations of ∑chlordanes were negatively correlated with red blood cell counts, hemoglobin, and hematocrit, indicative of anemia. Positive correlations were observed between most classes of OC contaminants and white blood cell counts and between mirex and ∑TCDD-like PCB concentrations and the heterophil:lymphocyte ratio, suggesting modulation of the immune system. All classes of OCs in the blood except dieldrin were correlated positively with aspartate aminotransferase (AST) activity, indicating possible hepatocellular damage. Mirex and ∑TCDD-like PCB blood concentrations were negatively correlated with alkaline phosphatase (ALP) activity. Significant correlations to levels of certain OC contaminant classes also suggested possible alteration of protein (↑blood urea nitrogen, ↓albumin:globulin ratio), carbohydrate (↓glucose), and ion (↑sodium, ↓magnesium) regulation. These correlations suggest that OC contaminants may be affecting the health of loggerhead sea turtles even though sea turtles accumulate lower concentrations of OCs compared with other wildlife.
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              Contaminant-induced immunotoxicity in harbour seals: wildlife at risk?

              Persistent, lipophilic polyhalogenated aromatic hydrocarbons (PHAHs) accumulate readily in the aquatic food chain and are found in high concentrations in seals and other marine mammals. Recent mass mortalities among several marine mammal populations have been attributed to infection by morbilliviruses, but a contributing role for immunotoxic PHAHs, including the polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs), and polychlorinated dibenzofurans (PCDFs) was not ruled out. We addressed this issue by carrying out a semi-field study in which captive harbour seals were fed herring from either the relatively uncontaminated Atlantic Ocean or the contaminated Baltic Sea for 2 years. We present here an overview of results obtained during this study. An impairment of natural killer (NK) cell activity, in vitro T-lymphocyte function, antigen-specific in vitro lymphocyte proliferative responses, and in vivo delayed-type hypersensitivity and antibody responses to ovalbumin was observed in the seals fed the contaminated Baltic herring. Additional feeding studies in PVG rats using the same herring batches suggested that an effect at the level of the thymus may be responsible for changes in cellular immunity, that virus-specific immune responses may be impaired, and that perinatal exposure to environmental contaminants represents a greater immunotoxic threat than exposure as a juvenile or adult. Together with the pattern of TCDD toxic equivalents of different PHAHs in the herring, these data indicate that present levels of PCBs in the aquatic food chain are immunotoxic to mammals. A review of contaminant levels in free-ranging harbour seals inhabiting polluted areas of Europe and North America suggests that many populations may be at risk to immunotoxicity. This could result in diminished host resistance and an increased incidence and severity of infectious disease.
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                Author and article information

                Journal
                Environ Health Perspect
                Environmental Health Perspectives
                National Institute of Environmental Health Sciences
                0091-6765
                January 2006
                21 September 2005
                : 114
                : 1
                : 70-76
                Affiliations
                [1 ] Nicholas School of the Environment and Earth Sciences, Coastal Systems Science and Policy, and Integrated Toxicology Program, Duke University, Beaufort, North Carolina, USA
                [2 ] National Institute of Standards and Technology, Hollings Marine Laboratory, Charleston, South Carolina, USA
                [3 ] Department of Environmental and Molecular Toxicology, and Center for Marine Science and Technologies, North Carolina State University, Morehead City, North Carolina, USA
                [4 ] Marine Biomedicine and Environmental Science Center, and
                [5 ] Department of Pediatrics, Medical University of South Carolina, Charleston, South Carolina, USA
                [6 ] Grice Marine Laboratory, College of Charleston, Charleston, South Carolina, USA
                [7 ] Mystic Aquarium and Institute for Exploration, Mystic, Connecticut, USA
                Author notes
                Address correspondence to J.M. Keller, National Institute of Standards and Technology, Hollings Marine Laboratory, 331 Ft. Johnson Rd., Charleston, SC 29412 USA. Telephone: (843) 762-8863. Fax: (843) 762-8742. E-mail: Jennifer.keller@noaa.gov
                Current address: University of Nevada-Las Vegas, Clinical Laboratory Sciences Program, Las Vegas, NV, USA.

                The authors declare they have no competing financial interests.

                Article
                ehp0114-000070
                10.1289/ehp.8143
                1332659
                16393661
                f1eea3b5-ddca-425a-acf2-6ae17cf63928
                This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original DOI.
                History
                : 23 March 2005
                : 21 September 2005
                Categories
                Research

                Public health
                polychlorinated biphenyls,pcbs,reptile,ddt,organochlorine pesticides,persistent organic pollutants,immunotoxicity,organochlorine contaminants

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