<!DOCTYPE article PUBLIC "-//NLM//DTD JATS (Z39.96) Journal Archiving and Interchange DTD v1.0 20120330//EN" "JATS-archivearticle1.dtd">
<article xmlns:xlink="http://www.w3.org/1999/xlink">
  <front>
    <journal-meta />
    <article-meta>
      <title-group>
        <article-title>Impacts of High Concentration of CO2 on the Serum Biochemistry and Carbonic Anhydrase Enzyme Activity of Rainbow Trout, Oncorhynchus mykiss</article-title>
      </title-group>
      <contrib-group>
        <contrib contrib-type="author">
          <string-name>Sevdan Yılmaz</string-name>
          <email>sevdanyilmaz@comu.edu.tr</email>
          <xref ref-type="aff" rid="aff0">0</xref>
        </contrib>
        <contrib contrib-type="author">
          <string-name>Hasan Kaya</string-name>
          <xref ref-type="aff" rid="aff1">1</xref>
        </contrib>
        <contrib contrib-type="author">
          <string-name>Mert Gürkan</string-name>
          <xref ref-type="aff" rid="aff2">2</xref>
        </contrib>
        <contrib contrib-type="author">
          <string-name>Olcay Hisar</string-name>
          <xref ref-type="aff" rid="aff1">1</xref>
        </contrib>
        <contrib contrib-type="author">
          <string-name>Kahraman Selvi</string-name>
          <xref ref-type="aff" rid="aff4">4</xref>
        </contrib>
        <contrib contrib-type="author">
          <string-name>Selçuk Türel</string-name>
          <xref ref-type="aff" rid="aff0">0</xref>
        </contrib>
        <contrib contrib-type="author">
          <string-name>Bilal Aydın</string-name>
          <xref ref-type="aff" rid="aff3">3</xref>
        </contrib>
        <contrib contrib-type="author">
          <string-name>Samet Çetin</string-name>
          <xref ref-type="aff" rid="aff3">3</xref>
        </contrib>
        <aff id="aff0">
          <label>0</label>
          <institution>Department of Aquaculture, Faculty of Marine Sciences and Technology, University of Çanakkale Onsekiz Mart</institution>
          ,
          <country country="TR">Turkey</country>
        </aff>
        <aff id="aff1">
          <label>1</label>
          <institution>Department of Basic Science, Faculty of Marine Sciences and Technology, University of Çanakkale Onsekiz Mart</institution>
          ,
          <country country="TR">Turkey</country>
        </aff>
        <aff id="aff2">
          <label>2</label>
          <institution>Department of Biology, Faculty of Art and Science, University of Çanakkale Onsekiz Mart</institution>
          ,
          <country country="TR">Turkey</country>
        </aff>
        <aff id="aff3">
          <label>3</label>
          <institution>Natural and Applied Sciences, University of Çanakkale Onsekiz Mart</institution>
          ,
          <country country="TR">Turkey</country>
        </aff>
        <aff id="aff4">
          <label>4</label>
          <institution>Yenice Vocational Collage, University of Çanakkale Onsekiz Mart</institution>
          ,
          <country country="TR">Turkey</country>
        </aff>
      </contrib-group>
      <fpage>695</fpage>
      <lpage>698</lpage>
      <abstract>
        <p>In this study, the physiological effects that the increasing carbondioxide concentrations on rainbow trout, Oncorhynchus mykiss is evaluated by using serum biochemical variables and carbonic anhydrase activities. The fish were exposed for 14 days to 14 mg/L concentrations of CO2. The serum GLU value showed a significant increase in the group exposed to CO2 compared to the control group at days 14. Serum TRI, COL and blood CA values showed a significant decrease in the group exposed to CO2 at day 7 compared to the control group. The TRI value a statistically significant increase in the group exposed to CO2 at day 14 compared to the control group. In conclusion, this study results indicate that the some serum biochemical variables and blood carbonic anhydrase activity of rainbow trout significantly affected by high level of CO2.</p>
      </abstract>
      <kwd-group>
        <kwd>Oncorhynchus mykiss</kwd>
        <kwd>carbondioxide</kwd>
        <kwd>blood</kwd>
        <kwd>carbonic anhydrase</kwd>
      </kwd-group>
    </article-meta>
  </front>
  <body>
    <sec id="sec-1">
      <title>1 Introduction</title>
      <p>
        It is now recognized that the 21st century will show a significant global warming
trend induced by an increase in atmospheric greenhouse gases
        <xref ref-type="bibr" rid="ref3">(Houghton et al.,
2001)</xref>
        . Carbon dioxide (CO2), one of the important green gases, has increased by
40% from pre-industrial levels from approximately 280 parts per million by volume
(ppmv) in the 18th century to 390 ppmv in 2010
        <xref ref-type="bibr" rid="ref4">(IPCC 2007)</xref>
        . Water sources are
attractive sites for possible storage of CO2. Addition of CO2 to the water will result
in a decrease in pH due to the bicarbonate buffer system in sea- and fresh-water. It is
supposed that disposal of sufficient CO2 to stabilize atmospheric levels at twice the
pre-industrial level by the end of this century would lower the pH of the entire water
sources on average by more than 0.1 units
        <xref ref-type="bibr" rid="ref1">(Caldeira and Wicket 2003)</xref>
        . Addition of
CO2 to the water will result in a decrease in pH due to the bicarbonate buffer system
in seawater and freshwater. This is a large fraction of the normal variation of pH in
open water sources. Research interest in CO2-driven water acidification has been
centred on certain groups of calcifying water organisms, but knowledge on the
possible impacts of water acidification on fish is limited.
      </p>
      <p>The purpose of the present study was to evaluate the impact of high water CO2
levels (14 mg/L) in freshwater on serum biochemical variables and carbonic
anhydrase activities in rainbow trout, Oncorhynchus mykiss for 14 days.
2</p>
    </sec>
    <sec id="sec-2">
      <title>Material and Method</title>
      <p>The experiment was designed in triplicate and 12 fish were placed in each
experimental tank (140 L). During the experiment, the fish were exposed for 14 days
to 14 mg/L concentrations of CO2 by injecting CO2 (purity 99.9%) gas by means of
ceramic diffusers. Control group was not exposed to CO2. In the experiment, five
fish from each aquarium on the 7th and 14th day were used for analysis. The serum
biochemical variables (glucose, total protein, albumin, triglyceride and cholesterol)
in the blood serum was measured according to Yılmaz et al (in press). The CO2
hydratase activity of the CA enzyme was assayed colorimetrically by using the
method of Wilbur and Anderson (1976). Each value was expressed as mean ±
standard error (SE) for each parameter measured. Student's t-test was used to
determine the significance of differences between the exposure group and control
group. The statistical analyses were carried out by using SPSS 17.0, and the
significance level was considered to be 0.05.</p>
    </sec>
    <sec id="sec-3">
      <title>3 Results</title>
      <p>In the present study, results (Table 1) showed that CO2 exposed group did not
show differences of Tprot, ALB and GLO values at any of the two sampling periods
as compared with the control group (P&gt;0.05). However, the serum GLU value
showed a significant increase in the group exposed to CO2 compared to the control
group at days 14 (P&lt;0.05). Serum TRI, COL and blood CA values showed a
significant decrease in the group exposed to CO2 at day 7 compared to the control
group (P&lt;0.05). The TRI value a statistically significant increase in the group
exposed to CO2 at day 14 compared to the control group (P&lt;0.05).</p>
    </sec>
    <sec id="sec-4">
      <title>4 Conclusion</title>
      <p>7 th day
14 th day</p>
      <p>Control</p>
      <p>
        Measurement of blood parameters can indicate the welfare status of fish
        <xref ref-type="bibr" rid="ref5">(Roncarati et al 2006)</xref>
        . The CO2 reactions within the RBC are catalyzed by carbonic
anhydrase (CA)
        <xref ref-type="bibr" rid="ref7">(Swenson and Maren 1987)</xref>
        . The rapid anion exchange mechanism
therefore facilitates the loading of CO2 into the blood at the tissue level and provides
plasma HCO3¯ with access to CA during the short period that blood passes through
the gills
        <xref ref-type="bibr" rid="ref2">(Currie et al 1995)</xref>
        . As a result of the study, it is identified that CO2
concentrations cause negative effects on the serum glucose, triglyceride, cholesterol
and blood carbonic anhydrase activity. In conclusion, this study results indicate that
the some serum biochemical variables and blood carbonic anhydrase activity of
rainbow trout significantly affected by high level of CO2.
      </p>
      <p>Acknowledgments. This work was partially supported by The Scientific and
Technological Council of Turkey (TUBITAK, Project Number: 113O220,
coordinated by H Kaya).</p>
    </sec>
  </body>
  <back>
    <ref-list>
      <ref id="ref1">
        <mixed-citation>
          1.
          <string-name>
            <surname>Caldeira</surname>
            ,
            <given-names>K.</given-names>
          </string-name>
          and
          <string-name>
            <surname>Wickett</surname>
            ,
            <given-names>M.E.</given-names>
          </string-name>
          (
          <year>2003</year>
          )
          <article-title>Oceanography: anthropogenic carbon and ocean pH</article-title>
          ,
          <source>Nature</source>
          ,
          <volume>425</volume>
          ,
          <fpage>365</fpage>
          .
        </mixed-citation>
      </ref>
      <ref id="ref2">
        <mixed-citation>
          2.
          <string-name>
            <surname>Currie</surname>
            ,
            <given-names>S.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Kieffer</surname>
            ,
            <given-names>J.D.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Tufts</surname>
            ,
            <given-names>B.L.</given-names>
          </string-name>
          (
          <year>1995</year>
          )
          <article-title>The effects of blood CO2 reactionrates on CO2 removal from muscle in exercised trout</article-title>
          .
          <source>Respiratory physiology</source>
          ,
          <volume>100</volume>
          :
          <fpage>261</fpage>
          -
          <lpage>269</lpage>
          .
        </mixed-citation>
      </ref>
      <ref id="ref3">
        <mixed-citation>
          3.
          <string-name>
            <surname>Houghton</surname>
            ,
            <given-names>J.H.</given-names>
          </string-name>
          et al. (
          <year>2001</year>
          )
          <article-title>Climate change 2001</article-title>
          .
          <article-title>The scientific basis, the contribution of working group I to the third assesment report of the intergovernment panel on climate change</article-title>
          . J.H.
          <string-name>
            <surname>Houghton</surname>
          </string-name>
          et al. (ed), Cambridge, Cambridge University Press.. 944 pp.
        </mixed-citation>
      </ref>
      <ref id="ref4">
        <mixed-citation>
          4. IPCC. (
          <year>2007</year>
          )
          <article-title>Summary for policymakers</article-title>
          .
          <source>in climate change</source>
          <year>2007</year>
          :
          <article-title>The physical science basis. working group I contribution to the fourth assessment report of the IPCC, edited by S</article-title>
          . Solomon,
          <string-name>
            <given-names>D.</given-names>
            <surname>Qin</surname>
          </string-name>
          ,
          <string-name>
            <given-names>M.</given-names>
            <surname>Manning</surname>
          </string-name>
          ,
          <string-name>
            <given-names>Z.</given-names>
            <surname>Chen</surname>
          </string-name>
          ,
          <string-name>
            <given-names>M.</given-names>
            <surname>Marquis</surname>
          </string-name>
          ,
          <string-name>
            <surname>K.B. Averyt</surname>
            ,
            <given-names>M.</given-names>
          </string-name>
          <string-name>
            <surname>Tignor</surname>
            , and
            <given-names>H.L.</given-names>
          </string-name>
          <string-name>
            <surname>Miller</surname>
          </string-name>
          ,
          <volume>1</volume>
          -
          <fpage>18</fpage>
          . Cambridge: Cambridge University Press.
        </mixed-citation>
      </ref>
      <ref id="ref5">
        <mixed-citation>
          5.
          <string-name>
            <surname>Roncarati</surname>
            ,
            <given-names>A.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Melotti</surname>
            ,
            <given-names>P.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Dees</surname>
            ,
            <given-names>A.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Mordenti</surname>
            ,
            <given-names>O.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Angellotti</surname>
            ,
            <given-names>L.</given-names>
          </string-name>
          (
          <year>2006</year>
          )
          <article-title>Welfare status of cultured sea bass (Dicentrarchus labrax L.) and seabream (Sparus aurata L.) assessed by blood parameters and tissue characteristics</article-title>
          .
          <source>Journal of applied ichthyology</source>
          ,
          <volume>22</volume>
          , p.
          <fpage>225</fpage>
          -
          <lpage>234</lpage>
        </mixed-citation>
      </ref>
      <ref id="ref6">
        <mixed-citation>
          6.
          <string-name>
            <surname>Sabine</surname>
            ,
            <given-names>C.L.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Christopher</surname>
            <given-names>L.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Feely</surname>
            ,
            <given-names>R.A.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Gruber</surname>
            ,
            <given-names>N.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Key</surname>
            ,
            <given-names>M.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Lee</surname>
            ,
            <given-names>K.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Bullister</surname>
            ,
            <given-names>J.L.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Wanninkhof</surname>
            ,
            <given-names>R.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Wong</surname>
            ,
            <given-names>C. S.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Wallace</surname>
            ,
            <given-names>D.W. R.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Tilbrook</surname>
            ,
            <given-names>B.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Millero</surname>
            ,
            <given-names>F.J.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Peng</surname>
            ,
            <given-names>T.H.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Kozyr</surname>
            ,
            <given-names>A.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Ono</surname>
            ,
            <given-names>T.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Rios</surname>
            ,
            <given-names>A.F.</given-names>
          </string-name>
          (
          <year>2004</year>
          )
          <article-title>The oceanic sink for anthropogenic CO2</article-title>
          .
          <source>Science</source>
          ,
          <volume>305</volume>
          , Issue 5682.
        </mixed-citation>
      </ref>
      <ref id="ref7">
        <mixed-citation>
          7.
          <string-name>
            <surname>Swenson</surname>
            ,
            <given-names>E.R.</given-names>
          </string-name>
          , and
          <string-name>
            <surname>Maren</surname>
            ,
            <given-names>T.H.</given-names>
          </string-name>
          (
          <year>1987</year>
          )
          <article-title>Roles of Gill and Red Cell Carbonic Anhydrase in Elasmobranch HCO3 and CO2 Excretion</article-title>
          .
          <article-title>The american</article-title>
          journal of physiology,
          <volume>253</volume>
          , p.
          <fpage>450</fpage>
          -
          <lpage>458</lpage>
          .
        </mixed-citation>
      </ref>
      <ref id="ref8">
        <mixed-citation>
          8.
          <string-name>
            <surname>Wilbur</surname>
            ,
            <given-names>K.M.</given-names>
          </string-name>
          , and
          <string-name>
            <surname>Anderson</surname>
            ,
            <given-names>N.G.</given-names>
          </string-name>
          (
          <year>1976</year>
          )
          <article-title>Electrometric and colorimetric determination of carbonic anhydrase</article-title>
          .
          <source>The journal of biological chemistry</source>
          ,
          <volume>176</volume>
          , p.
          <fpage>147</fpage>
          -
          <lpage>154</lpage>
          .
        </mixed-citation>
      </ref>
      <ref id="ref9">
        <mixed-citation>
          9.
          <string-name>
            <surname>Yılmaz</surname>
            ,
            <given-names>S.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Ergün</surname>
            ,
            <given-names>S.</given-names>
          </string-name>
          ,
          <string-name>
            <surname>Çelik</surname>
            ,
            <given-names>E.Ş.</given-names>
          </string-name>
          <article-title>(in press) Effect of dietary spice supplementations on welfare status of sea bass</article-title>
          ,
          <source>Dicentrarchus labrax L. Proceedings of the national academy of sciences, India section B: biological sciences.</source>
        </mixed-citation>
      </ref>
    </ref-list>
  </back>
</article>