Abstract
Anemia is a widespread hematological disorder in vertebrates. In channel catfish (Ictalurus punctatus), anemia is a persistent problem in culture environments, however, its causes and physiological impacts are not well understood. To better understand the symptoms and characterize associated biomarkers of anemia, 1-D 1H and 2-D 1H J-resolved NMR were used to analyze metabolite changes in healthy and anemic channel catfish kidney and liver tissue. Additionally, standard physiological analytical techniques were used to analyze blood plasma. NMR analyses revealed energy sources such as glucose were depleted and many metabolites associated with anaerobic metabolism or alternative energy pathways such as lactate, creatine, alanine, acetate and myo-inositol had changed. Energy demanding processes were reduced, such as muscle function, as indicated by reductions in taurine and inosine, and protein synthesis. Stress and oxidative stress related metabolites changed, with increases in valine, leucine and isoleucine and decreases in glutathione. Inhibitory neurotransmitters such as 4-aminobutyric acid (GABA) increased, and excitatory neurotransmitters such as glutamine and glutamate generally decreased, although there were tissue-specific differences. Immune function also decreased in anemic fish. Blood analyses revealed decreased respiratory gas transport capabilities through reductions in erythrocytes and hemoglobin-markers. Taken together, these tissues and analytical techniques produced complementary results, and metabolite expression in anemic channel catfish was similar to that observed in other vertebrates exposed to anoxia, severe hypoxia or experiencing oxidative stress from deficiencies in iron homeostasis.
References
Andras, F., & Meretey, K. (1992). Histamine: An early messenger in inflammatory and immune reactions. Immunology Today, 13, 154–156.
Andrews, J. W., & Murai, T. (1979). Pyridoxine requirements of channel catfish. Journal of Nutrition, 109, 533–537.
Atzori, L., Xanthos, T., Barberini, L., Antonucci, R., Murgia, F., Lussu, M., et al. (2010). A metabolomic approach in an experimental model of hypoxia-reoxygenation in newborn piglets: urine predicts outcome. The Journal of Maternal-Fetal and Neonatal Medicine, 23, 134–137.
Bower, R. H., Kern, K. A., & Fischer, J. E. (1985). Use of a branched chain amino acid enriched solution in patients under metabolic stress. The American Journal of Surgery, 149, 266–270.
Brown, D. W., Jack, S. W., & Waterstrat, P. R. (1994). Histopathology of hematopoietic tissue in channel catfish, Ictalurus punctatus, with anemia. Journal of Applied Aquaculture, 3, 101–108.
Butterworth, C. E, Jr, Plumb, J. A., & Grizzle, J. M. (1986). Abnormal folate metabolism in feed-related anemia of cultured channel catfish. Proceedings of the Society for Experimental Biology and Medicine, 181, 49–58.
Camus, A. C., Wise, D. J., Khoo, L. H., Shi, L. H., & Berghaus, R. D. (2014). Iron status of channel catfish Ictalurus punctatus affected by channel catfish anemia and response to parenteral iron. Diseases of Aquatic Organisms, 107, 241–248.
Clauss, T. M., Dove, A. D. M., & Arnold, J. E. (2008). Hematologic disorders of fish. Veterinary Clinics Exotic Animal Practice, 11, 445–462.
Dave, G., Johansson-Sjobeck, M. L., Larsson, A., Lewander, K., & Lidman, U. (1975). Metabolic and hematological effects of starvation in the European eel, Anguilla anguilla L.—I. Carbohydrate, lipid, protein and inorganic ion metabolism. Comparative Biochemistry and Physiology A, 52, 423–430.
De Bandt, J. P., & Cynober, L. (2006). Therapeutic use of branched-chain amino acids in burn, trauma, and sepsis. Journal of Nutrition, 136, 308S–313S.
Duncan, P. L., & Lovell, R. T. (1994). Influence of vitamin C on the folate requirement of channel catfish, Ictalurus punctatus, for growth, hematopoiesis, and resistance to Edwardsiella ictaluri infection. Aquaculture, 127, 233–244.
Duncan, P. L., Lovell, R. T., Butterworth, C. E, Jr, Freeberg, L. E., & Tamura, T. (1993). Dietary folate requirement determined for channel catfish, Ictalurus punctatus. Journal of Nutrition, 123, 1888–1897.
Feala, J. D., Coquin, L., McCulloch, A. D., & Paternostro, G. (2007). Flexibility in energy metabolism supports hypoxia tolerance in Drosophila flight muscle: Metabolomic and computational systems analysis. Molecular Systems Biology, 3, 99.
Fiehn, O., Roberston, D., Griffin, J., Werf, M., Nikolau, B., Morrison, N., et al. (2007). The metabolomics standards initiative (MSI). Metabolomics, 3, 175–178.
Fijan, N. (2002). Morphogenesis of blood cell lineages in channel catfish. Journal of Fish Biology, 60, 999–1014.
Goodpaster, A. M., Romick-Rosendale, L. E., & Kennedy, M. A. (2010). Statistical significance analysis of nuclear magnetic resonance-based metabonomics data. Analytical Biochemistry, 401, 134–143.
Grabitzki, J., & Lochnit, G. (2009). Immunomodulation by phosphocholine-biosynthesis, structures and immunological implications of parasitic PC-epitopes. Molecular Immunology, 47, 149–163.
Hallman, T. M., Rojas-Vargas, A. C., Jones, D. R., & Richards, J. G. (2008). Differential recovery from exercise and hypoxia exposure measured using 31P- and 1H-NMR in white muscle of the common carp Cyprinus carpio. Journal of Experimental Biology, 211, 3237–3248.
Hand, S. C., & Hardewig, I. (1996). Downregulation of cellular metabolism during environmental stress: mechanisms and implications. Annual Review of Physiology, 58, 539–563.
Hasko, G., Sitkovsky, M. V., & Szabo, C. (2004). Immunomodulatory and neuroprotective effects of inosine. Trends in Pharmacological Science, 25, 152–157.
Hu, X., Camus, A. C., Aono, S., Morrison, E. E., Dennis, J., Nusbaum, K. E., et al. (2007). Channel catfish hepcidin expression in infection and anemia. Comparative Immunology, Microbiology and Infectious Diseases, 30, 55–69.
Ill, A. M., Mitchell, T. R., Neely, E. B., & Connor, J. R. (2006). Metabolic analysis of mouse brains that have compromised iron storage. Metabolic Brain Disease, 21, 77–87.
Khoo, L. Diagnoses of anemia in channel catfish (unpublished data).
Klar, G. T., Hanson, L. A., & Brown, S. W. (1986). Diet related anemia in channel catfish: Case history and laboratory induction. The Progressive Fish-Culturalist, 48, 60–64.
Lardon, I., Eyckmans, M., Vu, T. N., Laukens, K., De Boek, G., & Dommisse, R. (2013a). 1H-NMR study of the metabolome of a moderately hypoxia-tolerant fish, the common carp (Cyprinus carpio). Metabolomics, 9, 1216–1227. doi:10.1007/s11306-013-0540-y.
Lardon, I., Nilsson, G. E., Stecyk, J. A. W., Vu, T. N., Laukens, K., Dommisse, R., & De Boek, G. (2013b). 1H-NMR study of the metabolome of an exceptionally anoxia tolerant vertebrate, the crucian carp (Carassius carassius). Metabolomics, 9, 311–323. doi:10.1007/s11306-012-0448-y.
Lee, R., West, D., Phillips, S. M., & Britz-McKibbin, P. (2010). Differential metabolomics for quantitative assessment of oxidative stress with strenuous exercise and nutritional intervention: Thiol-specific regulation of cellular metabolism with N-acetyl-l-cysteine pretreatment. Analytical Chemistry, 82, 2959–2968.
Li, P., Mai, K., Trushenski, J., & Wu, G. (2009). New developments in fish amino acid nutrition: Towards functional and environmentally oriented aquafeeds. Amino Acids, 37, 43–53.
Lin, C. Y., Wu, H., Tjeerdema, R. S., & Viant, M. R. (2007). Evaluation of metabolite extraction strategies from tissue samples using NMR metabolomics. Metabolomics, 3, 55–67.
Ludwig, C., Easton, J. M., Lodi, A., Tiziani, S., Manzoor, S., Southam, A. D., et al. (2012). Birmingham Metabolite Library: A publicly accessible database of 1-D 1H and 2D 1H J-resolved NMR spectra of authentic metabolite standards (BML-NMR). Metabolomics, 8, 8–18.
Ludwig, C., & Viant, M. R. (2010). Two-dimensional J-resolved NMR spectroscopy: Review of a key methodology in the metabolomics toolbox. Phytochemical Analysis, 21, 22–32.
Lujan, R., Shigemoto, R., & Lopez-Bendito, G. (2005). Glutamate and GABA receptor signalling in the developing brain. Neuroscience, 130, 567–580.
Lutz, P. L., Nilsson, G. E., & Perez-Pinzon, M. A. (1996). Anoxia tolerant animals from a neurobiological perspective. Comparative Biochemistry and Physiology, 113B, 3–13.
Miller, L. L., Bly, C. G., Watson, M. L., & Bale, W. F. (1951). The dominant role of the liver in plasma protein synthesis. Journal of Experimental Medicine, 94, 431–453.
Milligan, C. L. (1997). The role of cortisol in amino acid mobilizataion and metabolism following exhaustive exercise in rainbow trout (Oncorhynchus mykiss Walbaum). Fish Physiology and Biochemistry, 16, 119–128.
Minhas, G., Modgil, S., & Anand, A. (2014). Role of iron in ischemia-induced neurodegeneration: Mechanisms and insights. Metab Brain Disease, 1–9, 583–591. doi:10.1007/s11011-014-9522-7.
Noyes, A. D., Grizzle, J. M., & Plumb, J. A. (1991). Hematology and histopathology of an idiopathic anemia of channel catfish. Journal of Aquatic Animal Health, 3, 161–167.
Parsons, H. M., Ekman, D. R., Collette, T. W., & Viant, M. R. (2009). Spectral relative standard deviation: A practical benchmark in metabolomics. Analyst, 134, 478–485.
Pauli, G. F., Godecke, T., Jaki, B. U., & Lankin, D. C. (2012). Quantitative 1H NMR. Development and potential of an analytical method: An update. Journal of Natural Products, 75, 834–851.
Pena-Llopis, S., Ferrando, M. D., & Pena, J. B. (2003). Fish tolerance to organophosphate-induced oxidative stress is dependent on the glutathione metabolism and enhanced by N-acetylcysteine. Aquatic Toxicology, 65, 337–360.
Plumb, J. A., Horowitz, S. A., & Rogers, W. A. (1986). Feed-related anemia in cultured channel catfish (Ictalurus punctatus). Aquaculture, 51, 175–179.
Plumb, J. A., Liu, P. R., & Butterworth, C. E, Jr. (1994). Folic acid and pteroic acid related to anemia of cultured channel catfish, Ictalurus punctatus. Journal of Applied Aquaculture, 3, 91–100.
Podrabsky, J. E., Lopez, J. P., Fan, T. W. M., Higashi, R., & Somero, G. N. (2007). Extreme anoxia tolerance in embryos of the annual killifish Austrofundulus limnaeus: Insights from a metabolomics analysis. Journal of Experimental Biology, 210, 2253–2266.
Rao, R., Ennis, K., Oz, G., Lubach, G. R., Georgieff, M. K., & Coe, C. L. (2013). Metabolomic analysis of cerebrospinal fluid indicates iron deficiency compromises cerebral energy metabolism in the infant monkey. Neurochemical Research, 38, 573–580.
Rorabacher, D. (1991). Statistical treatment for rejection of deviant values: Critical values of Dixon’s Q parameter and related subrange ratios at the 95 % confidence level. Analytical Chemistry, 63, 139–146.
Saccenti, E., Hoefsloot, H. C. J., Smilde, A. K., Westerhuis, J. A., & Hendriks, M. M. W. B. (2013). Reflections on univariate and multivariate analysis of metabolomics data. Metabolomics. doi:10.1007/s11306-013-0598-6.
Schaffer, S. W., Jong, C. J., Ramila, K. C., & Azuma, J. (2010). Physiological roles of taurine in heart and muscle. Journal of Biomedical Science, 17(Suppl 1), S2.
Smith, C. E. (1968). Hematological changes in coho salmon fed a folic acid deficient diet. Journal of the Fisheries Research Board of Canada, 25, 151–156.
Soga, T., et al. (2006). Differential metabolomics reveals ophthalmic acid as an oxidative stress biomarker indicating hepatic glutathione consumption. Journal of Biological Chemistry, 281, 16768–16776.
Sumner, L. W., Amber, A., Barrett, D., Beale, M. H., Beger, R., Daykin, C. A., et al. (2007). Proposed minimum reporting standards for chemical analysis. Chemical Analysis Working Group (CAWG) Metabolomics Standard Initiative (MSI). Metabolomics, 3, 211–221.
Tavares, R. G., Santos, C. E. S., Tasca, C. I., Wajner, M., Souza, D. O., & Dutra-Filho, C. S. (2000). Inhibition of glutamate uptake into synaptic vesicles of rat brain by the metabolites accumulating in maple syrup urine disease. Journal of the Neurological Sciences, 181, 44–49.
Teague, C. R., Dhabhar, F. S., Barton, R. H., Beckwith-Hall, B., Powell, J., Cobain, M., et al. (2007). Metabonomic studies on the physiological effects of acute and chronic psychological stress in Sprague-Dawley rats. Journal of Proteome Research, 6, 2080–2093.
Tucker, C. S., Francis-Floyd, R., & Beleau, M. H. (1989). Nitrite-induced anemia in channel catfish, Ictalurus punctatus Rafinesque. Bulletin of Environmental Contamination and Toxicology, 43, 295–301.
Tuffnail, W., Mills, G. A., Cary, P., & Greenwood, R. (2009). An environmental 1H NMR metabolomic study of the exposure of the marine mussel Mytilus edulis to atrazine, lindane, hypoxia and starvation. Metabolomics, 5, 33–43.
Vijayan, M. M., Pereira, C., Grau, E. G., & Iwama, G. K. (1997). Metabolic responses associated with confinement stress in tilapia: The role of cortisol. Comparative Biochemistry and Physiology Part C: Pharmacology Toxicology and Endocrinology, 116, 89–95.
Ward, K. L., Tkac, I., Jing, Y., Felt, B., Beard, J., Connor, J., et al. (2007). Gestational and lactational iron deficiency alters the developing striatal metabolome and associated behaviors in young rats. Journal of Nutrition, 137, 1043–1049.
Weljie, A. M., Bondareva, A., Zang, P., & Jirik, F. R. (2011). 1H NMR metabolomics identification of markers of hypoxia-induced metabolic shifts in a breast cancer model system. Journal of Biomolecular NMR, 49, 185–193.
Wishart, D. S., Jewison, T., Guo, A. C., Wilson, M., Knox, C., Liu, Y., et al. (2013). HMDB 3.0—The human metabolome database in 2013. Nucleic Acids Research, 41, D801–D807.
Wu, H., Southam, A. D., Hines, A., & Viant, M. R. (2008). High-throughput tissue extraction protocol for NMR- and MS-based metabolomics. Analytical Biochemistry, 372, 204–212.
Yuk, J., McIntyre, K. L., Fischer, C., Hicks, J., Colson, K. L., Lui, E., et al. (2013). Distinguishing Ontario ginseng landraces and ginseng species using NMR-based metabolomics. Analytical and Bioanalytical Chemistry, 405, 4499–4509.
Yuk, J., McKelvie, J. R., Simpson, M. J., Spraul, M., & Simpson, A. J. (2010). Comparison of 1-D and 2-D NMR techniques for screening earthworm responses to sub-lethal endosulfan exposure. Environmental Chemistry, 7, 524–536.
Acknowledgments
We thank Todd Byars and Charles Walker for their technical assistance, the Mississippi Agricultural and Forestry Experiment Station, the US Department of Agriculture (USDA) Agricultural Research Service and the USDA National Institute of Food and Agriculture for funding and facility support; and two anonymous reviewers for helpful comments on the manuscript.
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The authors declare that they have no conflict of interest.
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All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.
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Allen, P.J., Wise, D., Greenway, T. et al. Using 1-D 1H and 2-D 1H J-resolved NMR metabolomics to understand the effects of anemia in channel catfish (Ictalurus punctatus). Metabolomics 11, 1131–1143 (2015). https://doi.org/10.1007/s11306-014-0767-2
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DOI: https://doi.org/10.1007/s11306-014-0767-2