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Isolated and combined exposure to ammonia and nitrite in giant freshwater pawn (Macrobrachium rosenbergii): effects on the oxidative stress, antioxidant enzymatic activities and apoptosis in haemocytes

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Abstract

The residual contaminators such as ammonia and nitrite are widely considered as relevant sources of aquatic environmental pollutants, posing a great threat to shrimp survival. To study the toxicological effects of ammonia and nitrite exposure on the innate immune response in invertebrates, we investigated the oxidative stress and apoptosis in haemocytes of freshwater prawn (Macrobrachium rosenbergii) under isolated and combined exposure to ammonia and nitrite in order to provide useful information about adult prawn immune responses. M. rosenbergii (13.44 ± 2.75 g) were exposed to 0, 5, and 25 mg/L total ammonia-N (TAN) and 0, 5, and 20 mg/L nitrite-N for 24 h. All ammonia concentrations were combined with all nitrite concentrations, making a total of nine treatments studied. Following the exposure treatment, antioxidant enzyme activity, reactive oxygen species (ROS) generation, nitric oxide (NO) generation, and apoptotic cell ratio of haemocytes were measured using flow cytometry. Results indicated that ROS generation was sensitive to the combined effect of ammonia and nitrite, which subsequently affected the Cu–Zn SOD activity. In addition, CAT showed the highest activity at 5 mg/L TAN while GPx decreased at 5 mg/L TAN and returned towards baseline at 25 mg/L. NO generation synchronized with the apoptotic cell ratio in haemocytes, indicating that NO production was closely associated with programmed cell death. Both NO production and apoptotic ratios significantly decreased following 25 mg/L TAN, which may be due to the antagonistic regulation of NO and GPx. We hypothesized that the toxicological effect of nitrite exhibited less change in physiological changes compared to that of ammonia, because of the high tolerance to nitrite exposure in mature M. rosenbergii and/or the competitive effects of chloride ions. Taken together, these results showed that ammonia and nitrite caused a series of combined oxidative stress and apoptosis in M. rosenbergi, but further studies are of great need to explain the mechanisms.

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References

  • Alcaraz G, Chiappa-Carrara X, Vanegas C (1997) Temperature tolerance of Penaeus setiferus postlarvae exposed to ammonia and nitrite. Aquat Toxicol 39:345–353

    Article  CAS  Google Scholar 

  • Armstrong DA, Stephenson MJ, Knight AW (1976) Acute toxicity of nitrite to larvae of the giant Malaysian prawn, Macrobrachium rosenbergii. Aquaculture 9:39–46

    Article  CAS  Google Scholar 

  • Armstrong DA, Chippendale D, Knight AW, Colt JE (1978) Interaction of ionized and un-ionized ammonia on short-term survival and growth of prawn larvae, macrobrachium rosenbergh. Biol Bull 154:15–31

    Article  CAS  Google Scholar 

  • Babior BM (1999) NADPH oxidase: an update. Blood 93:1464–1476

    CAS  Google Scholar 

  • Borutaite V, Brown GC (2006) S-nitrosothiol inhibition of mitochondrial complex I causes a reversible increase in mitochondrial hydrogen peroxide production. Biochimica et Biophysica Acta (BBA)-Bioenergetics 1757:562–566

    Article  CAS  Google Scholar 

  • Brown GC (2010) Nitric oxie and neuronal death. Nitric Oxide 23:153–165

    Article  CAS  Google Scholar 

  • Cavalli RO, Berghe EV, Lavens P, Thuy NTT, Wille M, Sorgeloos P (2000) Ammonia toxicity as a criterion for the evaluation of larval quality in the prawn Macrobrachium rosenbergii. Comp Biochem Physiol C 125:333–343

    CAS  Google Scholar 

  • Chanock SJ, El Benna J, Smith RM, Babior BM (1994) The respiratory burst oxidase. J Biol Chem 270:24519

    Google Scholar 

  • Chen JC, Cheng SY (1995) Accumulation of urea in the haemolymph and ammonia excretion of Penaeus japonicas exposed to ambient nitrite. Comp Biochem Physiol C 110:1–6

    Google Scholar 

  • Chen JC, Lee Y (1997) Effects of nitrite on mortality, ion regulation and acid-base balance of Macrobrachium rosenbergii at different external chloride concentrations. Aquat Toxicol 39:291–305

    Article  Google Scholar 

  • Cheng W, Chen JC (2002) The virulence of Enterococcus to freshwater prawn Macrobrachium rosenbergii and its immune resistance under ammonia stress. Fish Shellfish Immunol 12:97–109

    Article  Google Scholar 

  • Cheng W, Chen SM, Wang FI, Hsu PI, Liu CH, Chen JC (2003) Effects of temperature, pH, salinity and ammonia on the phagocytic activity and clearance efficiency of giant freshwater prawn Macrobrachium rosenbergii to Lactococcus garvieae. Aquaculture 219:111–121

    Article  Google Scholar 

  • Cheng W, Tung YH, Liu CH, Chen JC (2006) Molecular cloning and characterisation of cytosolic manganese superoxide dismutase (cytMn-SOD) from the giant freshwater prawn Macrobrachium rosenbergii. Fish Shellfish Immunol 20:438–449

    Article  CAS  Google Scholar 

  • Cheng SY, Shieh LW, Chen JC (2013) Changes in hemolymph oxyhemocyanin, acid–base balance, and electrolytes in Marsupenaeus japonicas under combined ammonia and nitrite stress. Aquat Toxicol 130:132–138

    Article  CAS  Google Scholar 

  • Chiu ST, Hsieh SL, Yeh SP, Jian SJ, Cheng W, Liu CH (2010) The increase of immunity and disease resistance of the giant freshwater prawn, Macrobrachium rosenbergii by feeding with selenium enriched-diet. Fish Shellfish Immunol 29:623–629

    Article  CAS  Google Scholar 

  • Cooper CE, Giulivi C (2007) Nitric oxide regulation of mitochondrial oxygen consumption II: molecular mechanism and tissue physiology. Am J Physiol-Cell Ph 292:C1993–C2003

    Article  CAS  Google Scholar 

  • Dandapat J, Chainy GBN, Janardhana Rao K (2003) Lipid peroxidation and antioxidant defence status during larval development and metamorphosis of giant prawn, Macrobrachium rosenbergii. Comp Biochem Physiol C 135:221–233

    Article  CAS  Google Scholar 

  • Das K, Samanta L, Chainy GBN (2000) A modified spectrophotometric assay of superoxide dismutase using nitrite formation by superoxide radicals. Indian J Biochem Bio 37:201–204

    CAS  Google Scholar 

  • Du J, Zhu HX, Liu P, Chen J, Xiu YJ, Yao W, Wu T, Ren Q, Meng QG, Gu W (2013) Immune responses and gene expression in hepatopancreas from Macrobrachium rosenbergii challenged by a novel pathogen spiroplasma MR-1008. Fish Shellfish Immunol 34:315–323

    Article  CAS  Google Scholar 

  • Guo H, Xian JA, Li B, Ye CX, Wang AL, Miao YT, Liao SA (2013) Gene expression of apoptosis-related genes, stress protein and antioxidant enzymes in hemocytes of white shrimp Litopenaeus vannamei under nitrite stress. Comp Biochem Physiol C 157:366–371

    CAS  Google Scholar 

  • Kim JH, Guo X, Park HS (2008) Comparison study of the effects of temperature and free ammonia concentration on nitrification and nitrite accumulation. Process Biochem 43:154–160

    Article  CAS  Google Scholar 

  • Kleinert H, Pautz A, Linker K, Schwarz PM (2004) Regulation of the expression of inducible nitric oxide synthase. Eur J Pharmacol 500:255–266

    Article  CAS  Google Scholar 

  • Liao Shaoan, Li Qing, Wang Anli, Xian Jianan, Chen Xiaodan, Gou Nina, Zhang Shengpeng, Wang Liang, Xuerun Xu (2012) Effect of nitrite on immunity of the white shrimp Litopenaeus vannamei at low temperture and low salinity. Ecotoxicology 21:1603–1608

    Article  CAS  Google Scholar 

  • Liu ZS, Li SM (2013) China fisheries yearbook. China Agriculture Press, Beijing

    Google Scholar 

  • Meinelt T, Kroupova H, Stüber A, Rennert B, Wienke A, Steinberg CEW (2010) Can dissolved aquatic humic substances reduce the toxicity of ammonia and nitrite in recirculating aquaculture systems? Aquaculture 306:378–383

    Article  CAS  Google Scholar 

  • Naqvi AA, Adhikari S, Pillai BR, Sarangi N (2007) Effect of ammonia-N on growth and feeding of juvenile Macrobrachium rosenbergii (De-Man). Aquac Res 38:847–851

    Article  CAS  Google Scholar 

  • Orrenius S, Gogvadze V, Zhivotovsky B (2007) Mitochondrial oxidative stress: implications for cell death. Annu Rev Pharmacol Toxicol 47:143–183

    Article  CAS  Google Scholar 

  • Palumbo A (2005) Nitric oxide in marine invertebrates: a comparative perspective. Comp Biochem Physiol A 142:241–248

    Article  CAS  Google Scholar 

  • Savvides SN, Scheiwein M, Böhme CC, Arteel GE, Karplus PA, Becker K, Schirmer RH (2002) Crystal structure of the antioxidant enzyme glutathione reductase inactivated by peroxynitrite. J Biol Chem 277:2779–2784

    Article  CAS  Google Scholar 

  • Tacon AGJ, Cody JJ, Conquest LD, Divakaran S, Forster LP, Decamp OE (2002) Effect of culture system on the nutrition and growth performance of pacific white shrimp Litopenaeus vannamei (Boone) fed different diets. Aquacult Nutr 8:121–137

    Article  Google Scholar 

  • Tidwell JH, Coyle S, Arnum A, Weibel C (2000) Production response of freshwater prawns Macrobrachium rosenbergiito increasing amounts of artificial substrate in ponds. J World Aquacult Soc 31:452–458

    Article  Google Scholar 

  • Vedel NE, Korsgaard B, Jensen FB (1998) Isolated and combined exposure to ammonia and nitrite in rainbow trout (Oncorhynchus mykiss): effects on electrolyte status, blood respiratory properties and brain glutamine/glutamate concentrations. Aquat Toxicol 41:325–342

    Article  CAS  Google Scholar 

  • Wang WN, Zhou J, Wang P, Tian TT, Zheng Y, Liu Y, Mai WJ, Wang AL (2009) Oxidative stress, DNA damage and antioxidant enzyme gene expression in the Pacific white shrimp, Litopenaeus vannamei when exposed to acute pH stress. Comp Biochem Physiol C 150:428–435

    Google Scholar 

  • Xian JA, Wang AL, Ye CX, Chen XD, Wang WN (2010) Phagocytic activity, respiratory burst, cytoplasmic free-Ca2+ concentration and apoptotic cell ratio of haemocytes from the black tiger shrimp, Penaeus monodon under acute copper stress. Comp Biochem Physiol C 152:182–188

    Google Scholar 

  • Xian JA, Wang AL, Chen XD, Gou NN, Miao YT, Liao SA, Ye CX (2011) Cytotoxicity of nitrite on haemocytes of the tiger shrimp, Penaeus monodon, using flow cytometric analysis. Aquaculture 317:240–244

    Article  CAS  Google Scholar 

  • Xian JA, Wang AL, Hao XM, Miao YT, Li B, Ye CX, Liao SA (2012) In vitro toxicity of nitrite on haemocytes of the tiger shrimp, Penaeus monodon, using flow cytometric analysis. Comp Biochem Physiol C 156:75–79

    CAS  Google Scholar 

  • Yeh SP, Liu KF, Chiu ST, Jian SJ, Cheng W, Liu CH (2009) Identification and cloning of a selenium dependent glutathione peroxidase from giant freshwater prawn, Macrobrachium rosenbergii. Fish Shellfish Immunol 27:181–191

    Article  CAS  Google Scholar 

  • Zbigniew T, Pokora Wojciech (2006) Individual and combined effect of anthracene, cadmium, and chloridazone on growth and activity of SOD izoformes in three Scenedesmus species. Ecotoxicol Environ Saf 65:323–331

    Article  CAS  Google Scholar 

  • Zhang SP, Li JF, Wu XC, Zhong WJ, Xian JA, Liao SA, Miao YT, Wang AL (2013) Effects of different dietary lipid level on the growth, survival and immune-relating genes expression in Pacific white shrimp, Litopenaeus vannamei. Fish Shellfish Immunol 34:1131–1138

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This research was supported by the National Natural Science Foundation of China (31100296), Guangdong Provincial Natural Science Foundation (S2011020003256), Scientific and Technological Planning Project of Guangdong Province (2012B020307004), and the Scientific and Technological Planning Project of Guangzhou City.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Key Laboratory of Ecology and Environmental Science in Guangdong Higher Education, Key Laboratory of Safe and Healthy Aquaculture in Guangdong Province, College of Life Science, South China Normal University, Guangzhou, 510631, People’s Republic of China

    Yufan Zhang, Chaoxia Ye, Anli Wang, Xuan Zhu, Changhong Chen, Jianan Xian & Zhenzhu Sun

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  1. Yufan Zhang
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  2. Chaoxia Ye
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  3. Anli Wang
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  4. Xuan Zhu
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  7. Zhenzhu Sun
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Correspondence to Chaoxia Ye or Anli Wang.

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Zhang, Y., Ye, C., Wang, A. et al. Isolated and combined exposure to ammonia and nitrite in giant freshwater pawn (Macrobrachium rosenbergii): effects on the oxidative stress, antioxidant enzymatic activities and apoptosis in haemocytes. Ecotoxicology 24, 1601–1610 (2015). https://doi.org/10.1007/s10646-015-1477-x

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