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<article xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:mml="http://www.w3.org/1998/Math/MathML" dtd-version="3.0" article-type="research-article" xml:lang="pt">
  <front>
    <journal-meta>
      <journal-id journal-id-type="publisher-id">Arq. Bras. Cardiol.</journal-id>
      <journal-title-group>
        <journal-title>Arquivos Brasileiros de Cardiologia</journal-title>
        <abbrev-journal-title abbrev-type="publisher">Arq. Bras.
					Cardiol.</abbrev-journal-title>
      </journal-title-group>
      <issn pub-type="ppub">0066-782X</issn>
      <issn pub-type="epub">1678-4170</issn>
      <publisher>
        <publisher-name>Sociedade Brasileira de Cardiologia</publisher-name>
      </publisher>
    </journal-meta>
    <article-meta>
      <article-id pub-id-type="publisher-id" specific-use="scielo-v3">Hczr8hPNwFtpxVTpkDkCSWM</article-id>
      <article-id pub-id-type="publisher-id" specific-use="scielo-v2">S0066-782X2014000100014</article-id>
      <article-id pub-id-type="other">0014</article-id>
      <article-id pub-id-type="doi">10.5935/abc.20130239</article-id>
      <article-categories>
        <subj-group subj-group-type="heading">
          <subject>Review Article</subject>
        </subj-group>
      </article-categories>
      <title-group>
        <article-title xml:lang="pt">A ecocardiografia no laborat&#243;rio cardiovascular: um
					guia para a pesquisa com animais</article-title>
      </title-group>
      <contrib-group>
        <contrib contrib-type="author">
          <name>
            <surname>Abduch</surname>
            <given-names>Maria Cristina Donadio</given-names>
          </name>
          <xref ref-type="aff" rid="aff1"/>
          <xref ref-type="corresp" rid="c01"/>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Assad</surname>
            <given-names>Renato Samy</given-names>
          </name>
          <xref ref-type="aff" rid="aff1"/>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Mathias</surname>
            <given-names>Wilson</given-names>
            <suffix>Jr</suffix>
          </name>
          <xref ref-type="aff" rid="aff1"/>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Aiello</surname>
            <given-names>Vera Demarchi</given-names>
          </name>
          <xref ref-type="aff" rid="aff1"/>
        </contrib>
        <aff id="aff1"><institution content-type="orgdiv2">Instituto do Cora&#231;&#227;o</institution><institution content-type="orgdiv1">Faculdade de Medicina</institution><institution content-type="orgname">Universidade de S&#227;o Paulo</institution><addr-line><named-content content-type="city">S&#227;o Paulo</named-content><named-content content-type="state">SP</named-content></addr-line><country>Brasil</country>Instituto do Cora&#231;&#227;o, Faculdade de Medicina da
					Universidade de S&#227;o Paulo, S&#227;o Paulo, SP - Brasil</aff>
      </contrib-group>
      <author-notes>
        <corresp id="c01"><bold>Correspond&#234;ncia:</bold> Maria Cristina Donadio Abduch, Pra&#231;a Guido
					Cagnacci, 05, Vila Madalena. CEP 05444-060, S&#227;o Paulo, SP - Brasil. E-mail:
						<email>cristinaabduch@cardiol.br</email>, <email>abduchmc@gmail.com</email>
				</corresp>
        <fn fn-type="conflict">
          <p>
            <bold>Potencial Conflito de Interesses</bold>
          </p>
          <p>Declaramos n&#227;o haver conflito de interesses pertinentes.</p>
        </fn>
      </author-notes>
      <pub-date pub-type="collection">
        <month>1</month>
        <year>2014</year>
      </pub-date>
      <pub-date pub-type="epub">
        <day>14</day>
        <month>12</month>
        <year>2013</year>
      </pub-date>
      <volume>102</volume>
      <issue>1</issue>
      <fpage>97</fpage>
      <lpage>103</lpage>
      <history>
        <date date-type="received">
          <day>02</day>
          <month>04</month>
          <year>2013</year>
        </date>
        <date date-type="rev-recd">
          <day>24</day>
          <month>07</month>
          <year>2013</year>
        </date>
        <date date-type="accepted">
          <day>29</day>
          <month>07</month>
          <year>2013</year>
        </date>
      </history>
      <permissions>
        <license license-type="open-access" xlink:href="http://creativecommons.org/licenses/by-nc/3.0/">
          <license-p>This is an Open Access article distributed under the terms of the
						Creative Commons Attribution Non-Commercial License which permits
						unrestricted non-commercial use, distribution, and reproduction in any
						medium, provided the original work is properly cited. </license-p>
        </license>
      </permissions>
      <abstract xml:lang="pt">
        <p>A exequibilidade e o potencial para investigar o cora&#231;&#227;o, sob o ponto de vista
					morfol&#243;gico e hemodin&#226;mico, v&#234;m aumentando a aplica&#231;&#227;o da ecocardiografia na
					pesquisa. Al&#233;m disso, o desenvolvimento de novas tecnologias, tais como a
					ecocardiografia tridimensional em tempo real e o speckle tracking, requerem
					valida&#231;&#227;o por meio de estudos experimentais antes de serem institu&#237;das no
					cen&#225;rio cl&#237;nico.</p>
        <p>O presente artigo objetiva fornecer informa&#231;&#245;es acerca das particularidades do
					estudo ecocardiogr&#225;fico nos mam&#237;feros quadr&#250;pedes, com vistas &#224; pesquisa
					experimental.</p>
      </abstract>
      <kwd-group xml:lang="pt">
        <kwd>Ecocardiografia/diagn&#243;stico</kwd>
        <kwd>Ecocardiografia/tend&#234;ncias</kwd>
        <kwd>Experimenta&#231;&#227;o Animal</kwd>
        <kwd>Cora&#231;&#227;o/fisiopatologia</kwd>
      </kwd-group>
      <counts>
        <fig-count count="6"/>
        <table-count count="2"/>
        <equation-count count="0"/>
        <ref-count count="35"/>
        <page-count count="1"/>
      </counts>
    </article-meta>
  </front>
  <body>
    <sec sec-type="intro">
      <title>Introdu&#231;&#227;o</title>
      <p>Devido ao baixo custo, &#224; caracter&#237;stica n&#227;o invasiva, &#224; portabilidade e ao potencial
				para investigar o cora&#231;&#227;o sob o ponto de vista morfol&#243;gico e hemodin&#226;mico, a
				ecocardiografia vem ocupando um papel importante na pesquisa experimental e
					cl&#237;nica<xref ref-type="bibr" rid="B01">1</xref>.</p>
      <p>O m&#233;dico sueco Inge Edler, considerado o "Pai da Ecocardiografia", foi o primeiro a
				utilizar esse m&#233;todo em pesquisa experimental. No in&#237;cio dos anos 1950, Edler
				demonstrou, em cora&#231;&#245;es de bezerros, que os sinais ecocardiogr&#225;ficos que se
				acreditava serem provenientes da parede anterior do &#193;trio Esquerdo (AE) eram, na
				verdade, oriundos da c&#250;spide anterior da Valva Mitral (VM)<xref ref-type="bibr" rid="B02">2</xref>. Desde ent&#227;o, um n&#250;mero crescente de estudos experimentais
				envolvendo a ecocardiografia vem sendo publicados.</p>
      <p>Recentemente, a Ecocardiografia 3D em Tempo Real (E3DTR), o Doppler Tecidual (TDI) e
				o Speckle Tracking (STE) representam ferramentas tecnol&#243;gicas inovadoras no
				diagn&#243;stico precoce de dissincronia e de disfun&#231;&#227;o card&#237;aca diast&#243;lica e sist&#243;lica,
				trazendo &#224; luz do conhecimento cient&#237;fico novas aplica&#231;&#245;es cl&#237;nicas<xref ref-type="bibr" rid="B03">3</xref>
				<sup>,</sup>
				<xref ref-type="bibr" rid="B04">4</xref>. Apesar de a resson&#226;ncia magn&#233;tica card&#237;aca
				ser atualmente considerada o m&#233;todo de refer&#234;ncia para o c&#225;lculo dos volumes e da
				fra&#231;&#227;o de eje&#231;&#227;o ventricular, in&#250;meros par&#226;metros ecocardiogr&#225;ficos podem fornecer,
				na rotina cl&#237;nica, informa&#231;&#245;es acuradas das dimens&#245;es e dos volumes ventriculares,
				al&#233;m da fun&#231;&#227;o mioc&#225;rdica.</p>
      <p>O presente artigo tem por objetivo fazer uma revis&#227;o dos aspectos t&#233;cnicos da
				ecocardiografia no laborat&#243;rio experimental, considerando suas vantagens e suas
				limita&#231;&#245;es, de acordo com a esp&#233;cie animal.</p>
      <sec>
        <title>Planos da ecocardiografia transtor&#225;cica e modelo animal</title>
        <p>Atualmente, a investiga&#231;&#227;o pela Ecocardiografia Transtor&#225;cica (ETT) nos animais
					empregados em pesquisa experimental &#233; favorecida pelo desenvolvimento
					tecnol&#243;gico dos equipamentos; entretanto, s&#227;o essenciais a escolha do transdutor
					com frequ&#234;ncia adequada e o conhecimento das particularidades dos mam&#237;feros
					quadr&#250;pedes.</p>
        <p>De acordo com os princ&#237;pios f&#237;sicos, transdutores com baixa frequ&#234;ncia (2,0 a 3,5
					MHz) apresentam menor resolu&#231;&#227;o da imagem. No entanto, a penetra&#231;&#227;o do feixe de
					eco &#233; maior, o que permite seu uso em animais com superf&#237;cie corp&#243;rea
						elevada<xref ref-type="bibr" rid="B05">5</xref>. Inversamente, sondas com
					alta frequ&#234;ncia (4,0 a 8,0 MHz) n&#227;o t&#234;m grande penetra&#231;&#227;o, por&#233;m fornecem imagem
					com maior resolu&#231;&#227;o, sendo mais apropriadas para o uso em animais menores e/ou
					mais jovens<xref ref-type="bibr" rid="B06">6</xref>
					<sup>-</sup>
					<xref ref-type="bibr" rid="B08">8</xref>. Considerando coelhos e pequenos
					roedores (ratos, camundongos e cobaias), para se obter resolu&#231;&#227;o satisfat&#243;ria da
					imagem &#233; necess&#225;rio o uso o uso de transdutores com frequ&#234;ncias maiores (10,0 a
					15,0 MHz), em raz&#227;o do tamanho reduzido desses animais<xref ref-type="bibr" rid="B09">9</xref>
					<sup>,</sup>
					<xref ref-type="bibr" rid="B10">10</xref>. Assim, uma vez que a frequ&#234;ncia do
					transdutor &#233; inversamente proporcional &#224; penetra&#231;&#227;o do feixe de ultrassom,
					quanto menor o mam&#237;fero, maior a frequ&#234;ncia da sonda.</p>
        <p>Os mam&#237;feros quadr&#250;pedes apresentam o cora&#231;&#227;o localizado na por&#231;&#227;o m&#233;dia da caixa
					tor&#225;cica, com o &#225;pice voltado para a esquerda; o t&#243;rax geralmente apresenta
					formato em quilha, o que permite a obten&#231;&#227;o de imagens a partir de ambos os
					hemit&#243;rax - esquerdo e direito - conforme mostrado abaixo.</p>
        <p>Com o intuito de adquirir imagens com &#243;tima qualidade t&#233;cnica, o animal deve ter
					o t&#243;rax tricotomizado, evitando a presen&#231;a de ar entre o transdutor e a
					superf&#237;cie corp&#243;rea, j&#225; que isso representa uma barreira para a penetra&#231;&#227;o do
					feixe de eco. Al&#233;m disso, a seda&#231;&#227;o ou a anestesia s&#227;o cruciais para a acur&#225;cia
					do exame; a elei&#231;&#227;o da droga apropriada, na dose correta para cada esp&#233;cie, e o
					conhecimento de poss&#237;veis altera&#231;&#245;es no sistema cardiovascular tamb&#233;m devem ser
						considerados<xref ref-type="bibr" rid="B11">11</xref>.</p>
        <p>O posicionamento varia de acordo com o modelo animal: para c&#227;es e porcos, &#233;
					indicado o dec&#250;bito lateral esquerdo; coelhos e pequenos roedores devem ser
					mantidos em dec&#250;bito dorsal; enquanto que os ruminantes necessitam de uma maca
					customizada, pois apresentam o t&#243;rax extremamente aquilhado (<xref ref-type="fig" rid="f01">Figura 1</xref>).</p>
        <fig id="f01" fig-type="other">
          <label>Figura 1</label>
          <caption>
            <title>Maca customizada para animais com t&#243;rax aquilhado, permitindo a
							obten&#231;&#227;o dos planos ecocardiogr&#225;ficos.</title>
          </caption>
          <graphic xlink:href="1678-4170-abc-102-01-0097-30239-gf01.jpg"/></fig>
        <p>Geralmente, as aquisi&#231;&#245;es das imagens ecocardogr&#225;ficas s&#227;o as mesmas que para os
					humanos, &#224; exce&#231;&#227;o dos ruminantes, que n&#227;o apresentam abordagem apical. Por
					ocupar uma posi&#231;&#227;o medial no t&#243;rax, ter o &#225;pice apontando para a esquerda e para
					o diafragma, o cora&#231;&#227;o dos mam&#237;feros quadr&#250;pedes apresenta algumas
					peculiaridades. Uma delas &#233;, de fato, uma desvantagem: a impossibilidade da
					aquisi&#231;&#227;o de imagens de boa qualidade pela janela subcostal, uma vez que o
					&#226;ngulo de incid&#234;ncia n&#227;o &#233; perpendicular ao cora&#231;&#227;o (entre os ruminantes, isso &#233;
					exacerbado pela presen&#231;a dos pr&#233;-est&#244;magos). A outra &#233; a capacidade de se
					obterem imagens de ambos os lados do t&#243;rax.</p>
        <p>Segue-se uma descri&#231;&#227;o das particularidades da abordagem ecocardiogr&#225;fica, de
					acordo com o modelo animal. A <xref ref-type="table" rid="t01">Tabela 1</xref>
					oferece um guia resumido relacionado a cada um deles e a <xref ref-type="fig" rid="f02">Figura 2</xref> mostra a posi&#231;&#227;o do transdutor em rela&#231;&#227;o ao t&#243;rax
					do animal. &#201; importante salientar que, com rela&#231;&#227;o aos valores normais das
					estruturas card&#237;acas (di&#226;metros e volumes), as mesmas variam de acordo com o
					tamanho e a esp&#233;cie. No entanto, o ecocardiograma basal &#233; focado na an&#225;lise da
					fun&#231;&#227;o mioc&#225;rdica e da performance valvar antes do procedimento experimental,
					com a finalidade de atestar que o cora&#231;&#227;o dos animais pode ser considerado
					saud&#225;vel.</p>
        <table-wrap id="t01">
          <label>Tabela 1</label>
          <caption>
            <title>Guia r&#225;pido para o exame ecocardiogr&#225;fico em pesquisa cardiovascular
							com animais</title>
          </caption>
          <table rules="rows" frame="hsides">
            <thead>
              <tr>
                <td>
                  <bold>Animal</bold>
                </td>
                <td align="center">
                  <bold>Frequ&#234;ncia do transdutor (MHz)</bold>
                </td>
                <td align="center">
                  <bold>Posi&#231;&#227;o do animal</bold>
                </td>
                <td align="center">
                  <bold>Principais planos ecocardiogr&#225;ficos</bold>
                </td>
                <td align="center">
                  <bold>Principais limita&#231;&#245;es</bold>
                </td>
              </tr>
            </thead>
            <tbody>
              <tr style="border-bottom-style:hidden">
                <td>Roedores pequenos</td>
                <td rowspan="2" align="center">10,0 - 15,0</td>
                <td rowspan="2" align="center">Dec&#250;bito lateral esquerdo</td>
                <td rowspan="2" align="center">PLAX, eixo curto, apical</td>
                <td rowspan="2" align="center">Frequ&#234;ncia card&#237;aca
									elevada</td>
              </tr>
              <tr>
                <td>Coelhos</td>
              </tr>
              <tr style="border-bottom-style:hidden">
                <td rowspan="2">Cabras/ovelhas</td>
                <td align="center">2,5 - 3,5 (adultos)</td>
                <td align="center">Dec&#250;bito lateral esquerdo</td>
                <td rowspan="2" align="center">PLAX, eixo curto</td>
                <td rowspan="2" align="center">Formato do t&#243;rax</td>
              </tr>
              <tr>
                <td align="center">4,0 - 8,0 (jovens)</td>
                <td align="center">Maca personalizada</td>
              </tr>
              <tr style="border-bottom-style:hidden">
                <td rowspan="2">C&#227;es</td>
                <td align="center">2,5 - 3,5 (adultos)</td>
                <td rowspan="2" align="center">Dec&#250;bito lateral esquerdo</td>
                <td rowspan="2" align="center">PLAX, eixo curto, apical</td>
                <td rowspan="2" align="center">---</td>
              </tr>
              <tr>
                <td align="center"> 4,0 - 8,0 (jovens)</td>
              </tr>
              <tr style="border-bottom-style:hidden">
                <td rowspan="2">Porcos</td>
                <td align="center">2,5 - 3,5 (adultos)</td>
                <td rowspan="2" align="center">Dec&#250;bito lateral esquerdo</td>
                <td rowspan="2" align="center">PLAX, eixo curto, apical</td>
                <td rowspan="2" align="center">---</td>
              </tr>
              <tr>
                <td align="center">4,0 - 8,0 (jovens)</td>
              </tr>
            </tbody>
          </table>
          <table-wrap-foot>
            <fn>
              <p>PLAX: corte paraesternal eixo longo.</p>
            </fn>
          </table-wrap-foot>
        </table-wrap>
        <fig id="f02" fig-type="other">
          <label>Figura 2</label>
          <caption>
            <title>Posi&#231;&#227;o do transdutor em rela&#231;&#227;o ao t&#243;rax do animal na abordagem pelo
							hemit&#243;rax esquerdo (A) e direito (B).</title>
          </caption>
          <graphic xlink:href="1678-4170-abc-102-01-0097-30239-gf02.jpg"/></fig>
      </sec>
      <sec>
        <title>Janela paraesternal esquerda</title>
        <p>Poss&#237;vel de ser obtida em c&#227;es, porcos, coelhos e pequenos roedores, no terceiro
					espa&#231;o intercostal:</p>
        <p>- plano paraesternal longitudinal (<xref ref-type="fig" rid="f03">Figura
						3A</xref>): permite visibiliza&#231;&#227;o do Ventr&#237;culo Direito (VD), Ventr&#237;culo
					Esquerdo (VE), Via de Sa&#237;da do Ventr&#237;culo Esquerdo (VSVE), Aorta (Ao) e Valva
					A&#243;rtica (VAo), VM e AE.</p>
        <fig id="f03" fig-type="other">
          <label>Figura 3</label>
          <caption>
            <title>Planos ecocardiogr&#225;ficos obtidos a partir da janela parasternal
							esquerda em c&#227;o. Plano paraesternal longitudinal(A). Plano parasternal
							transversal no n&#237;vel da base (B e C). Plano longitudinal da via de
							entrada do ventr&#237;culo direito (D). VD: ventr&#237;culo direito; VE:
							ventr&#237;culo esquerdo; Ao: aorta; AE: &#225;trio esquerdo; AD: &#225;trio direito;
							TP: tronco pulmonar; APD: art&#233;ria pulmonar direita; APE: art&#233;ria
							pulmonar esquerda; VSVD: via de sa&#237;da do ventr&#237;culo direito.</title>
          </caption>
          <graphic xlink:href="1678-4170-abc-102-01-0097-30239-gf03.jpg"/></fig>
        <p>- plano paraesternal transversal: no n&#237;vel da base, podem-se observar a VAo ao
					centro, o VD, o &#193;trio Direito (AD), a Valva Tric&#250;spide (VT) e o AE (<xref ref-type="fig" rid="f03">Figura 3B</xref>). Direcionando a sonda para a
					cabe&#231;a do animal, &#233; poss&#237;vel obter imagens da Via de Sa&#237;da do Ventr&#237;culo Direito
					(VSVD), da Valva Pulmonar (VP) e do Tronco Pulmonar (TP), sendo que esse &#250;ltimo
					se bifurca nos ramos direito e esquerdo (<xref ref-type="fig" rid="f03">Figura
						3C</xref>). O eixo curto, ao n&#237;vel da VM e dos m&#250;sculos papilares,
					geralmente n&#227;o &#233; bem visibilizado nessa abordagem, devido ao posicionamento
					obl&#237;quo do cora&#231;&#227;o.</p>
        <p>- plano longitudinal da via de entrada do VD (<xref ref-type="fig" rid="f03">Figura 3D</xref>).</p>
      </sec>
      <sec>
        <title>Janela paraesternal direita</title>
        <p>Exequ&#237;vel em todos os modelos animais, no terceiro espa&#231;o intercostal. &#201;
					importante notar que, nos ruminantes, esta &#233; a &#250;nica abordagem ecocardiogr&#225;fica
					transtor&#225;cica acurada, proporcionando mensura&#231;&#245;es que n&#227;o seriam poss&#237;veis pelas
					janelas convencionais, como no caso do c&#225;lculo da massa ventricular direita,
					segundo o qual o plano longitudinal da via de entrada do VD substitui o plano
					apical 4-c&#226;maras<xref ref-type="bibr" rid="B12">12</xref>.</p>
        <p>No plano paraesternal longitudinal (<xref ref-type="fig" rid="f04">Figura
						4A</xref>), &#233; poss&#237;vel visibilizar as mesmas estruturas abordadas no plano
					paraesternal longitudinal esquerdo (Ao, AE, VE e VD), al&#233;m do AD.</p>
        <fig id="f04" fig-type="other">
          <label>Figura 4</label>
          <caption>
            <title>Planos ecocardiogr&#225;ficos obtidos a partir da janela parasternal
							direita em c&#227;o (A e C) e cabra (B e D). Plano paraesternal longitudinal
							(A). Plano longitudinal da via de entrada dos ventr&#237;culos esquerdo e
							direito (B). Plano paraesternal transversal no n&#237;vel da base (C) e dos
							m&#250;sculos papilares (D). MV: valva mitral; VE: ventr&#237;culo esquerdo; VD:
							ventr&#237;culo direito; Ao: aorta; AE: &#225;trio esquerdo; AD: &#225;trio
							direito.</title>
          </caption>
          <graphic xlink:href="1678-4170-abc-102-01-0097-30239-gf04.jpg"/></fig>
        <p>O plano longitudinal da via de entrada dos ventr&#237;culos esquerdo e direito (<xref ref-type="fig" rid="f04">Figura 4B</xref>) pode ser usado como substituto
					para o plano apical; entretanto, deve-se ter cuidado no que tange ao &#225;pice
					verdadeiro, o qual n&#227;o pode ser avaliado por essa abordagem.</p>
        <p>O plano parasternal transversal &#233; obtido nos n&#237;veis da base, VM (<xref ref-type="fig" rid="f04">Figura 4C</xref>) e m&#250;sculos papilares (<xref ref-type="fig" rid="f04">Figura 4D</xref>), dirigindo o transdutor da
					posi&#231;&#227;o mais anterior em rela&#231;&#227;o ao cora&#231;&#227;o (regi&#245;es basais, &#224; semelhan&#231;a do
					plano paraesternal transversal esquerdo, <xref ref-type="fig" rid="f03">Figura
						3B</xref>), passando pelo n&#237;vel da VM, at&#233; chegar a uma posi&#231;&#227;o mais
					posterior, na altura dos m&#250;sculos papilares (com o transdutor voltado para os
					membros posteriores do animal).</p>
      </sec>
      <sec>
        <title>Janela apical</title>
        <p>Poss&#237;vel de ser obtida em c&#227;es, porcos, coelhos e pequenos roedores, no quinto
					espa&#231;o intercostal do hemit&#243;rax esquerdo, na regi&#227;o do <italic>ictus
						cordis</italic> (<xref ref-type="fig" rid="f05">Figura 5</xref>). Esse plano
					&#233; essencial para executar a an&#225;lise pelo TDI, uma vez que a amostra do Doppler
					deve estar paralela &#224; parede mioc&#225;rdica<xref ref-type="bibr" rid="B13">13</xref>.</p>
        <fig id="f05" fig-type="other">
          <label>Figura 5</label>
          <caption>
            <title>Planos apicais adquiridos em c&#227;es. Plano apical 4-c&#226;maras (A); plano
							apical 2-c&#226;maras (B); plano apical longitudinal (C). VE: ventr&#237;culo
							esquerdo; Ao: aorta; AE: &#225;trio esquerdo; AD: &#225;trio direito; VD:
							ventr&#237;culo direito.</title>
          </caption>
          <graphic xlink:href="1678-4170-abc-102-01-0097-30239-gf05.jpg"/></fig>
      </sec>
      <sec>
        <title>Ecocardiografia transesof&#225;gica</title>
        <p>A Ecocardiografia Transesof&#225;gica (ETE) no laborat&#243;rio experimental tem provado
					ser uma ferramenta acurada quando a ETT n&#227;o &#233; capaz de fornecer imagens
					adequadas, ou nos casos em que se faz necess&#225;rio o estudo das estruturas
					card&#237;acas durante um procedimento cir&#250;rgico<xref ref-type="bibr" rid="B14">14</xref>. &#201; exequ&#237;vel em grande parte dos modelos animais e, novamente, a
					droga utilizada na anestesia deve ser escolhida de acordo com a esp&#233;cie<xref ref-type="bibr" rid="B14">14</xref>
					<sup>-</sup>
					<xref ref-type="bibr" rid="B19">19</xref>.</p>
        <p>As imagens s&#227;o muito semelhantes &#224;quelas obtidas de humanos, sendo originadas do
					plano transg&#225;strico e de tr&#234;s planos adicionais dentro do es&#244;fago: cranial,
					m&#233;dio e caudal.</p>
      </sec>
      <sec>
        <title>Transg&#225;strico</title>
        <p>Esse plano &#233; bastante limitado, abordando apenas o VE no plano transversal.</p>
      </sec>
      <sec>
        <title>Es&#244;fago caudal</title>
        <p>O plano transversal fornece imagens dos ventr&#237;culos direito e esquerdo na altura
					da VM. Movimentos suaves de anteflex&#227;o permitem a abordagem da base do cora&#231;&#227;o,
					no n&#237;vel da VAo, semelhantemente ao plano paraesternal transversal na ETT.</p>
        <p>O plano longitudinal possibilita obter imagens do VE, VSVE e VM; rodando a sonda
					no sentido anti-hor&#225;rio, observa-se a via de entrada do VE, AD, septo
					interatrial e veias cavas.</p>
      </sec>
      <sec>
        <title>Es&#244;fago m&#233;dio</title>
        <p>- Plano 4-c&#226;maras (<xref ref-type="fig" rid="f06">Figura 6</xref>)</p>
        <fig id="f06" fig-type="other">
          <label>Figura 6</label>
          <caption>
            <title>Ecocardiografia transesof&#225;gica em c&#227;o. Note a biopr&#243;tese em posi&#231;&#227;o
							mitral. VE: ventr&#237;culo esquerdo; AE: &#225;trio esquerdo; AD: &#225;trio direito;
							VD: ventr&#237;culo direito.</title>
          </caption>
          <graphic xlink:href="1678-4170-abc-102-01-0097-30239-gf06.jpg"/></fig>
        <p>- Plano longitudinal: oferece imagens do AE, VE e Ao, como no plano apical
					longitudinal da ETT</p>
      </sec>
      <sec>
        <title>Es&#244;fago cranial</title>
        <p>- Plano transversal: arco a&#243;rtico, VSVE e valva pulmonar.</p>
        <p>- Plano longitudinal: o feixe centralizado permite a obten&#231;&#227;o de imagens da Ao
					ascendente e da VSVE; a rota&#231;&#227;o do transdutor no sentido hor&#225;rio fornece imagens
					da VSVD e do TP.</p>
      </sec>
      <sec>
        <title>Tecnologias inovadoras</title>
        <p>Nas &#250;ltimas d&#233;cadas, houve um desenvolvimento r&#225;pido e crescente de tecnologias
					inovadoras e, muitas vezes, essas ferramentas precisam ser testadas
					experimentalmente antes de serem introduzidas na pr&#225;tica cl&#237;nica.</p>
        <p>O contraste ecocardiogr&#225;fico tem por finalidade aumentar a defini&#231;&#227;o dos bordos
					endoc&#225;rdicos e, em associa&#231;&#227;o com a ecocardiografia sob estresse, pode ser
					empregado na pesquisa para a avalia&#231;&#227;o da perfus&#227;o mioc&#225;rdica<xref ref-type="bibr" rid="B20">20</xref>.</p>
        <p>A ecocardiografia tridimensional em tempo real, e o STE bidimensional e
					tridimensional representam ferramentas promissoras na avalia&#231;&#227;o e no diagn&#243;stico
					precoce de doen&#231;as card&#237;acas<xref ref-type="bibr" rid="B21">21</xref>
					<sup>-</sup>
					<xref ref-type="bibr" rid="B24">24</xref>. A valida&#231;&#227;o experimental dessas
					t&#233;cnicas foi muito importante para introduzi-las na pr&#225;tica cl&#237;nica,
					principalmente considerando o STE, uma vez que n&#227;o h&#225; um padr&#227;o-ouro verdadeiro
					que possa ser aplicado com essa finalidade em humanos: em rela&#231;&#227;o &#224; mec&#226;nica
					torsional, o elegante estudo de Helle Valle e cols.<xref ref-type="bibr" rid="B25">25</xref>, assim como o de Langeland e cols.<xref ref-type="bibr" rid="B26">26</xref>, Amundsen e cols.<xref ref-type="bibr" rid="B27">27</xref> e Seo e cols.<xref ref-type="bibr" rid="B28">28</xref>, que se
					referem ao strain mioc&#225;rdico bidimensional e tridimensional, a t&#233;cnica foi
					validada em compara&#231;&#227;o &#224; sonomicrometria, o que requer procedimento cir&#250;rgico a
					c&#233;u aberto para a inser&#231;&#227;o de cristais no mioc&#225;rdio. A <xref ref-type="table" rid="t02">Tabela 2</xref> mostra a correla&#231;&#227;o e o coeficiente de
					concord&#226;ncia entre o padr&#227;o-ouro e o STE nesses ensaios. Por&#233;m, para que se
					obtenham dados acurados, &#233; muito importante a utiliza&#231;&#227;o de anestesia
					inalat&#243;ria, para que seja induzida apneia respirat&#243;ria durante a aquisi&#231;&#227;o das
					imagens, uma vez que, tanto a E3DTR quanto o STE t&#234;m sua acur&#225;cia afetada pelos
					movimentos respirat&#243;rios do t&#243;rax. Outra quest&#227;o de grande relev&#226;ncia &#233; o uso da
					configura&#231;&#227;o adequada do equipamento, j&#225; que a resolu&#231;&#227;o temporal baixa,
					representada pela quantidade de quadros por segundo (<italic>frame
					rate</italic>), pode tamb&#233;m interferir nos resultados<xref ref-type="bibr" rid="B22">22</xref>.</p>
        <table-wrap id="t02">
          <label>Tabela 2</label>
          <caption>
            <title>Valida&#231;&#227;o experimental do speckle tracking por meio da
							sonomicrometria</title>
          </caption>
          <table rules="rows" frame="hsides">
            <thead>
              <tr style="border-bottom-style:hidden">
                <td>
                  <bold>Tipo do estudo</bold>
                </td>
                <td rowspan="2" align="center">
                  <bold>Par&#226;metro</bold>
                </td>
                <td rowspan="2" align="center">
                  <bold>Correla&#231;&#227;o</bold>
                </td>
              </tr>
              <tr>
                <td>
                  <bold> (Refer&#234;ncia #)</bold>
                </td>
              </tr>
            </thead>
            <tbody>
              <tr style="border-bottom-style:hidden">
                <td rowspan="3">2D STE (22)</td>
                <td align="center">Rota&#231;&#227;o apical</td>
                <td align="center">0,92</td>
              </tr>
              <tr style="border-bottom-style:hidden">
                <td align="center">Rota&#231;&#227;o basal</td>
                <td align="center">0,76</td>
              </tr>
              <tr>
                <td align="center">
                  <italic>Twist </italic>
                </td>
                <td align="center">0,94</td>
              </tr>
              <tr style="border-bottom-style:hidden">
                <td rowspan="2">2D STE (26)</td>
                <td align="center"><italic>Strain </italic>radial</td>
                <td align="center">0,72</td>
              </tr>
              <tr>
                <td><italic>Strain </italic>longitudinal</td>
                <td align="center"> 0,80</td>
              </tr>
              <tr>
                <td>2D STE (27)</td>
                <td align="center"><italic>Strain </italic>longitudinal</td>
                <td align="center">0,90</td>
              </tr>
              <tr>
                <td>3D STE (28)</td>
                <td align="center"><italic>Strain </italic>de &#225;rea</td>
                <td align="center">0,87</td>
              </tr>
            </tbody>
          </table>
          <table-wrap-foot>
            <fn>
              <p>2D STE- speckle tracking 2D; 3D STE- speckle tracking 3D.</p>
            </fn>
          </table-wrap-foot>
        </table-wrap>
      </sec>
    </sec>
    <sec sec-type="conclusions">
      <title>Considera&#231;&#245;es finais</title>
      <p>De acordo com o conhecimento dos autores, os primeiros estudos sistem&#225;ticos que
				empregaram a ecocardiografia no laborat&#243;rio experimental foram realizados nos anos
				1960. Lategola, em 1966, por meio de um transdutor de ultrassom miniaturizado,
				colocado no interior da aorta de c&#227;es, avaliou de forma invasiva o d&#233;bito card&#237;aco,
				num prot&#243;tipo do que seria, alguns anos depois, aplicado clinicamente pela ETT com o
				estudo Doppler de ondas cont&#237;nuas<xref ref-type="bibr" rid="B29">29</xref>. Dois
				anos depois, Christensen e Bonte mostraram, em c&#227;es, a acur&#225;cia da ecocardiografia
				na detec&#231;&#227;o de derrame peric&#225;rdico. Desde ent&#227;o, sua import&#226;ncia na investiga&#231;&#227;o
				experimental, com vistas &#224; aplica&#231;&#227;o cl&#237;nica, s&#243; tem aumentado<xref ref-type="bibr" rid="B30">30</xref>.</p>
      <p>Informa&#231;&#245;es relevantes, que n&#227;o seriam t&#227;o facilmente adquiridas com outras
				tecnologias, como a resson&#226;ncia nuclear magn&#233;tica, podem ser obtidas por meio da
				abordagem ecocardiogr&#225;fica. Segundo a experi&#234;ncia dos autores, o m&#233;todo &#233; &#250;til no
				seguimento da hipertrofia mioc&#225;rdica e de insufici&#234;ncia card&#237;aca<xref ref-type="bibr" rid="B12">12</xref>
				<sup>,</sup>
				<xref ref-type="bibr" rid="B31">31</xref>
				<sup>,</sup>
				<xref ref-type="bibr" rid="B32">32</xref>.</p>
      <p>O desenvolvimento de tecnologias inovadoras, como a biomicroscopia ultrass&#244;nica, que
				utiliza frequ&#234;ncias da ordem de 30 a 100 MHz, permite a an&#225;lise de estruturas
				min&#250;sculas - como o cora&#231;&#227;o fetal de camundongos no interior do &#250;tero materno<xref ref-type="bibr" rid="B33">33</xref>
				<sup>-</sup>
				<xref ref-type="bibr" rid="B35">35</xref>. Certamente isso ser&#225; de extrema
				import&#226;ncia no estudo do sistema cardiovascular e no desenvolvimento de
				microcirurgias em pequenos animais. Isso representa uma op&#231;&#227;o muito atraente em
				pesquisa experimental, uma vez que tais animais s&#227;o de f&#225;cil manipula&#231;&#227;o, apresentam
				melhor rela&#231;&#227;o custo-benef&#237;cio e podem ser mantidos em pequenas &#225;reas, viabilizando
				o estudo de grandes amostras num curto per&#237;odo de tempo.</p>
    </sec>
    <sec>
      <title>Contribui&#231;&#227;o dos autores</title>
      <p>Concep&#231;&#227;o e desenho da pesquisa: Abduch MCD, Aiello VD; Obten&#231;&#227;o de dados, An&#225;lise e
				interpreta&#231;&#227;o dos dados e Revis&#227;o cr&#237;tica do manuscrito quanto ao conte&#250;do
				intelectual: Abduch MCD, Assad RS, Aiello VD; Reda&#231;&#227;o do manuscrito: Abduch MCD,
				Assad RS, Mathias Jr. W, Aiello VD.</p>
    </sec>
  </body>
  <back>
    <ack>
      <p>Gostar&#237;amos de agradecer ao Rodrigo Tonan, pela assist&#234;ncia t&#233;cnica com as
				figuras.</p>
    </ack>
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    <fn-group>
      <fn fn-type="financial-disclosure">
        <p>
          <bold>Fontes de Financiamento</bold>
        </p>
        <p>O presente estudo n&#227;o teve fontes de financiamento externas.</p>
      </fn>
      <fn fn-type="other">
        <p>
          <bold>Vincula&#231;&#227;o Acad&#234;mica</bold>
        </p>
        <p>N&#227;o h&#225; vincula&#231;&#227;o deste estudo a programas de p&#243;s-gradua&#231;&#227;o.</p>
      </fn>
    </fn-group>
  </back>
  <sub-article id="S01" article-type="translation" xml:lang="en">
    <front-stub>
      <title-group>
        <article-title xml:lang="en">The echocardiography in the cardiovascular laboratory:
					a guide to research with animals</article-title>
      </title-group>
      <contrib-group>
        <contrib contrib-type="author">
          <name>
            <surname>Abduch</surname>
            <given-names>Maria Cristina Donadio</given-names>
          </name>
          <xref ref-type="aff" rid="aff2"/>
          <xref ref-type="corresp" rid="c02"/>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Assad</surname>
            <given-names>Renato Samy</given-names>
          </name>
          <xref ref-type="aff" rid="aff2"/>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Mathias</surname>
            <given-names>Wilson</given-names>
            <suffix>Jr</suffix>
          </name>
          <xref ref-type="aff" rid="aff2"/>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Aiello</surname>
            <given-names>Vera Demarchi</given-names>
          </name>
          <xref ref-type="aff" rid="aff2"/>
        </contrib>
        <aff id="aff2">
					Instituto do Cora&#231;&#227;o, Faculdade de Medicina da Universidade de S&#227;o Paulo, S&#227;o Paulo, SP - Brazil
				</aff>
      </contrib-group>
      <author-notes>
        <corresp id="c02"><bold>Mailing Address:</bold> Maria Cristina Donadio Abduch, Pra&#231;a Guido
					Cagnacci, 05, Vila Madalena. Postal Code 05444-060, S&#227;o Paulo, SP - Brazil.
					E-mail: <email>cristinaabduch@cardiol.br</email>,
						<email>abduchmc@gmail.com</email>
				</corresp>
        <fn fn-type="conflict">
          <p>
            <bold>Potential Conflict of Interest</bold>
          </p>
          <p>No potential conflict of interest relevant to this article was reported.</p>
        </fn>
      </author-notes>
      <history>
        <date date-type="received">
          <day>02</day>
          <month>04</month>
          <year>2013</year>
        </date>
        <date date-type="rev-recd">
          <day>24</day>
          <month>07</month>
          <year>2013</year>
        </date>
        <date date-type="accepted">
          <day>29</day>
          <month>07</month>
          <year>2013</year>
        </date>
      </history>
      <abstract xml:lang="en">
        <p>The feasibility and potential for the morphological and hemodynamic investigation
					of the heart has been increasing the use of the echocardiography in the research
					setting. Additionally, the development of new technologies, like the real time
					3D echocardiography and speckle tracking, demands validation throughout
					experimental studies before being instituted in the clinical setting.</p>
        <p>This paper aims to provide information concerning the particularities of the
					echocardiographic examination in quadruped mammals, targeting the experimental
					research.</p>
      </abstract>
      <kwd-group xml:lang="en">
        <kwd>Echocardiography / diagnosis</kwd>
        <kwd>Echocardiography / tend&#234;ncias</kwd>
        <kwd>Animal Experimentation</kwd>
        <kwd>Heart / physiopathology</kwd>
      </kwd-group>
      <counts>
        <fig-count count="6"/>
        <table-count count="2"/>
        <equation-count count="0"/>
        <ref-count count="35"/>
        <page-count count="1"/>
      </counts>
    </front-stub>
    <body>
      <sec sec-type="intro">
        <title>Introduction</title>
        <p>Considering the low cost and noninvasive nature, the portability and potential of
					morphological and hemodynamic investigation of the heart, the echocardiography
					plays an important role in the clinical and experimental research setting<xref ref-type="bibr" rid="B01">1</xref>. </p>
        <p>The Swedish physician Inge Edler, considered as "The Father of the
					Echocardiography", was the first to use this method in the experimental
					research. In the early 50's, Edler demonstrated in calf hearts that the echo
					signals, firstly thought to be generated by left atrial anterior wall, in fact
					were from the anterior mitral valve leaflet<xref ref-type="bibr" rid="B02">2</xref>. Since then, an increasing number of experimental studies using
					echocardiography has been published.</p>
        <p>Recently, Real Time 3D Echocardiography (RT3DE), Tissue Doppler Imaging (TDI) and
					Speckle Tracking (STE) represent novel technology tools in the early diagnosis
					of dyssynchrony, diastolic and systolic heart dysfunction, shedding light on
					scientific knowledge for clinical applications<xref ref-type="bibr" rid="B03">3</xref>
					<sup>,</sup>
					<xref ref-type="bibr" rid="B04">4</xref>. Although cardiac magnetic resonance
					imaging is currently considered the reference technique for ventricle volumetry
					and calculation of the ejection fraction, several echocardiographic parameters
					can provide reliable information on ventricular dimensions and volumes, besides
					myocardial function in daily clinical practice.</p>
        <p>This paper aims to review the technical aspects of echocardiography in the
					experimental laboratory, considering its advantages and limitations according to
					the animal species.</p>
        <sec>
          <title>The Transthoracic Echocardiography (TTE) Planes and Animal Model</title>
          <p>Nowadays, the transthoracic echocardiographic investigation in animals
						employed in experimental research is favored by the technological
						development of the equipment; however, it is crucial to select the right
						frequency transducer and have the knowledge on quadruped mammals
						peculiarities. </p>
          <p>According to physical principles, low-frequency transducers (2.0 - 3.5 MHz)
						offer smaller image resolution; nevertheless, the ultrasound beam
						penetration is enhanced, which allows their use in animals with a bigger
						body surface area<xref ref-type="bibr" rid="B05">5</xref>. Inversely, the
						high-frequency probes (4.0 - 8.0 MHz) do not enable larger penetrations,
						however, they are feasible for a better image resolution, being more
						adequate for smaller and/or younger animals<xref ref-type="bibr" rid="B06">6</xref>
						<sup>-</sup>
						<xref ref-type="bibr" rid="B08">8</xref>. Regarding rabbits and small
						rodents (rats, mice, and guinea pigs), in order to obtain a satisfactory
						image resolution, the use of higher frequency transducer (10.0 - 15.0 MHz)
						is needed, because of their very small size<xref ref-type="bibr" rid="B09">9</xref>
						<sup>,</sup>
						<xref ref-type="bibr" rid="B10">10</xref>. Therefore, once the frequency of
						the transducer is inversely proportional to the ultrasound beam penetration,
						the smaller the mammal, the greater the frequency of the probe.</p>
          <p>The heart of quadruped mammals present the heart located in the middle part
						of chest, with the apex pointed to the left; also, the chest is generally
						keel-shaped, allowing to obtain images from both left and right sides of the
						thorax, as shown below.</p>
          <p>In order to acquire images of optimal quality, the animal should have its fur
						chest removed, avoiding the presence of air between the transducer and the
						body surface, once it represents a barrier to the echo beam penetration.
						Besides that, sedation or anesthesia is crucial to the accuracy of the test;
						choosing the proper drug and dosage, according to the species, and the
						appraisal of possible alterations in the cardiovascular system should be
							scrutinized<xref ref-type="bibr" rid="B11">11</xref>. The positioning
						varies according to the animal model: for dogs and pigs, left lateral
						decubitus is indicated; rabbits and little rodents should be kept in dorsal
						decubitus while ruminants need a customized stretcher due to their extremely
						kneel-shaped chest. (<xref ref-type="fig" rid="f07">Figure 1</xref>)</p>
          <fig id="f07" fig-type="other">
            <label>Figure 1</label>
            <caption>
              <title>Customized stretcher for animals with kneel-shaped chest, in
								order to obtain the parasternal echocardiographic views.</title>
            </caption>
            <graphic xlink:href="1678-4170-abc-102-01-0097-30239-gf01-en.jpg"/></fig>
          <p>Usually, the echocardiographic image acquisitions are the same as in humans,
						except for the ruminants, which do not have the apical approach. The heart
						of quadruped mammals, which occupies a medial position in the chest, and the
						apex pointed to the left and the diaphragm, confers two echocardiographic
						peculiarities. One of them is indeed a disadvantage: the impossibility of
						acquiring images of good quality through the subcostal approach, once the
						angle of insonation is not perpendicular to the heart (among the ruminants
						this is exacerbated by the presence of the pre-stomachs). The other is the
						feasibility of obtaining images both by left and right hemithorax.</p>
          <p>Subsequently, a description of the approaches particularities according to
						the animal model; <xref ref-type="table" rid="t03">Table 1</xref> provides a
						summarized guide related to each one of them and <xref ref-type="fig" rid="f08">Figure 2</xref> depicts the transducer position in relation to
						the chest of the animal. Of note, regarding the normal values of heart
						structures (diameters and volumes), they vary according to the species and
						size; therefore, the baseline echocardiography is focused on the myocardial
						function and valvular performance analysis before the experimental
						procedure, in order to confirm that the heart of the animals can be
						considered healthy.</p>
          <table-wrap id="t03">
            <label>Table 1</label>
            <caption>
              <title>Rapid guide to the echocardiographic examination in animal
								cardiovascular research</title>
            </caption>
            <table rules="rows" frame="hsides">
              <thead>
                <tr>
                  <td>
                    <bold>Animal</bold>
                  </td>
                  <td align="center">
                    <bold>Transducer Frequency (MHz)</bold>
                  </td>
                  <td align="center">
                    <bold>Animal Position</bold>
                  </td>
                  <td align="center">
                    <bold>Main Echocardiographic Planes</bold>
                  </td>
                  <td align="center">
                    <bold>Main Limitations</bold>
                  </td>
                </tr>
              </thead>
              <tbody>
                <tr style="border-bottom-style:hidden">
                  <td>Small rodents</td>
                  <td rowspan="2" align="center">10.0 - 15.0</td>
                  <td rowspan="2" align="center">Left lateral decubitus</td>
                  <td rowspan="2" align="center">PLAX, short-axis, apical</td>
                  <td rowspan="2" align="center">Elevated heart rate</td>
                </tr>
                <tr>
                  <td>Rabbits</td>
                </tr>
                <tr style="border-bottom-style:hidden">
                  <td rowspan="2">Goats/Sheep</td>
                  <td align="center">2.5 - 3.5 (adultos)</td>
                  <td align="center">Left lateral decubitus</td>
                  <td rowspan="2" align="center">PLAX, short axis</td>
                  <td rowspan="2" align="center">Chest shape</td>
                </tr>
                <tr>
                  <td align="center">4.0 - 8.0 (jovens)</td>
                  <td align="center">Customized stretcher</td>
                </tr>
                <tr style="border-bottom-style:hidden">
                  <td rowspan="2">Dogs</td>
                  <td align="center">2.5 - 3.5 (adultos)</td>
                  <td rowspan="2" align="center">Left lateral decubitus</td>
                  <td rowspan="2" align="center">PLAX, short-axis, apical</td>
                  <td rowspan="2" align="center">---</td>
                </tr>
                <tr>
                  <td align="center"> 4.0 - 8.0 (jovens)</td>
                </tr>
                <tr style="border-bottom-style:hidden">
                  <td rowspan="2">Pigs</td>
                  <td align="center">2.5 - 3.5 (adultos)</td>
                  <td rowspan="2" align="center">Left lateral decubitus</td>
                  <td rowspan="2" align="center">PLAX, short-axis, apical</td>
                  <td rowspan="2" align="center">---</td>
                </tr>
                <tr>
                  <td align="center">4.0 - 8.0 (jovens)</td>
                </tr>
              </tbody>
            </table>
            <table-wrap-foot>
              <fn>
                <p>PLAX: paraesternal long-axis view.</p>
              </fn>
            </table-wrap-foot>
          </table-wrap>
          <fig id="f08" fig-type="other">
            <label>Figure 2</label>
            <caption>
              <title>Position of the transducer in relation to the chest of the animal
								in left (A) and right (B) hemithorax approach.</title>
            </caption>
            <graphic xlink:href="1678-4170-abc-102-01-0097-30239-gf02-en.jpg"/></fig>
        </sec>
        <sec>
          <title>Left parasternal approach</title>
          <p>Feasible to obtain in dogs, pigs, rabbits and little rodents, at the third
						intercostal space.</p>
          <p>Parasternal longitudinal plane (<xref ref-type="fig" rid="f09">Figure
							3A</xref>): allows visualization of the right ventricle (RV), left
						ventricle (LV), left ventricular outflow tract (LVOT), aorta (Ao) and aortic
						valve (AV), mitral valve (MV) and left atrium (LA).</p>
          <fig id="f09" fig-type="other">
            <label>Figure 3</label>
            <caption>
              <title>Echocardiographic views from the left parasternal approach in a
								dog. Parasternal long-axis view (A). Parasternal short-axis view at
								the basal level (B and C) Long-axis view of the right ventricular
								inflow tract (D). RV: right ventricle; LV: left ventricle; Ao:
								aorta; LA: left atrium; RA: right atrium; PT: pulmonary trunk; RPA:
								right pulmonary artery; LPA: left pulmonary artery.</title>
            </caption>
            <graphic xlink:href="1678-4170-abc-102-01-0097-30239-gf03-en.jpg"/></fig>
          <p>Parasternal short-axis view: at the basal level it is observed the AV in the
						middle, the RV, right atrium (RA), tricuspid valve (TV) and the LA (<xref ref-type="fig" rid="f09">Figure 3B</xref>). Tilting the probe towards
						the animal's head, it is possible to obtain images of the right ventricular
						outflow tract (RVOT), pulmonary valve (PV) and pulmonary trunk (PT), the
						latter winging in the right and left pulmonary branches (<xref ref-type="fig" rid="f09">Figure 3C</xref>). The short axes at the MV and
						papillary muscles level generally are not well visualized in this approach,
						because of the obliquely positioned heart.</p>
          <p>Long-axis view of the right ventricular inflow tract (<xref ref-type="fig" rid="f09">Figure 3D</xref>).</p>
        </sec>
        <sec>
          <title>Right parasternal approach</title>
          <p>Achievable in all animal models, at the third intercostal level. Of note,
						regarding the ruminants, this is the only reliable transthoracic
						echocardiography approach, allowing measurements that would not be feasible
						by the conventional views, as in the case of right ventricular mass
						calculation, where the long-axis view of the right ventricular inflow tract
						act as a surrogate for the apical 4-chamber view<xref ref-type="bibr" rid="B12">12</xref>. </p>
          <p>Parasternal longitudinal view (<xref ref-type="fig" rid="f10">Figure
							4A</xref>). In this view it is possible to visualize the same structures
						assessed through the left parasternal longitudinal approach (Ao, LA, LV and
						RV), besides the RA.</p>
          <fig id="f10" fig-type="other">
            <label>Figure 4</label>
            <caption>
              <title>Echocardiographic views from the right parasternal approach in a
								dog (A and C) and a goat (C and D). Parasternal long-axis view (A).
								Left and right ventricular inflow tract long-axis view (B).
								Parasternal short-axis view at the baseline (C) and at the papillary
								muscles level (D).MV: mitral valve; RV: right ventricle; LV: left
								ventricle; Ao: aorta; LA: left atrium; RA: right atrium.</title>
            </caption>
            <graphic xlink:href="1678-4170-abc-102-01-0097-30239-gf04-en.jpg"/></fig>
          <p>Left and right ventricular inflow tract long-axis view (<xref ref-type="fig" rid="f10">Figure 4B</xref>) As mentioned above, this view might be used
						as a substitute for the apical view; however, care must be taken when
						considering the true apical region, which cannot be evaluated by this
						approach. </p>
          <p>Parasternal short-axis view: obtained at the basal, MV (<xref ref-type="fig" rid="f10">Figure 4C</xref>), and papillary muscles level (<xref ref-type="fig" rid="f10">Figure 4D</xref>), by tilting the transducer
						from the most anterior position in relation to the heart (basal regions,
						similar to the left parasternal short-axis view, <xref ref-type="fig" rid="f09">Figure 3B</xref>), passing by the MV level to a most posterior
						one, in order to assess the papillary muscles (by tilting the transducer
						towards the animal's forelimbs). </p>
        </sec>
        <sec>
          <title>Apical approach</title>
          <p>Possible to obtain in dogs, pigs, rabbits and little rodents, at the fifth
						intercostal space, in the left hemithorax, at the <italic>ictus
							cordis</italic> (<xref ref-type="fig" rid="f11">Figure 5</xref>). This
						plane is essential to perform the TDI analysis, since the Doppler sample
						volume has to be parallel to the myocardial wall<xref ref-type="bibr" rid="B13">13</xref>.</p>
          <fig id="f11" fig-type="other">
            <label>Figure 5</label>
            <caption>
              <title>Apical views acquisition in a dog. A) Apical 4-chamber view; B)
								Apical 2-chamber view; C) Apical long-axis view. LV: left ventricle;
								Ao: aorta; LA: left atrium; RA: right atrium; RV: right
								ventricle.</title>
            </caption>
            <graphic xlink:href="1678-4170-abc-102-01-0097-30239-gf05-en.jpg"/></fig>
        </sec>
        <sec>
          <title>The Transesophageal Echocardiography (TEE)</title>
          <p>The TEE in the experimental lab is proving to be a reliable tool when TTE
						does not provide adequate images or when it is necessary to assess the heart
						structures during surgical procedure<xref ref-type="bibr" rid="B14">14</xref>. It is feasible in a variety of animal models and, again, the
						drug used to achieve anesthesia must be chosen according to the species<xref ref-type="bibr" rid="B14">14</xref>
						<sup>-</sup>
						<xref ref-type="bibr" rid="B19">19</xref>. </p>
          <p>The images are very similar to those obtained in humans, composed by a
						transgastric view and three additional views within the esophagus: cranial,
						middle and caudal. </p>
        </sec>
        <sec>
          <title>Transgastric</title>
          <p>This view is very limiting, only approaching the LV in the short axis
						view.</p>
        </sec>
        <sec>
          <title>Caudal Esophagus</title>
          <p>Short axis view: provides images of the right and left ventricles at the MV
						level; soft anti-flexion movements allow the approach of the heart base, at
						the AV level, similar to the parasternal short axis view in the TTE. </p>
          <p>Long axis view: in this plane it is possible to obtain images of the LV, LVOT
						and MV; rotating the probe counterclockwise allows imaging the left
						ventricular inflow tract, RA, atrial septum, and caval veins. </p>
        </sec>
        <sec>
          <title>Middle Esophagus</title>
          <p>Four-chamber view (<xref ref-type="fig" rid="f12">Figure 6</xref>).</p>
          <fig id="f12" fig-type="other">
            <label>Figure 6</label>
            <caption>
              <title>Transesophageal echocardiography in a dog. Note the bioprosthetic
								valve in the mitral valve position. LV: left ventricle; LA: left
								atrium; RA: right atrium; RV: right ventricle.</title>
            </caption>
            <graphic xlink:href="1678-4170-abc-102-01-0097-30239-gf06-en.jpg"/></fig>
          <p>Long axis view: provides imaging of the LA, LV and aorta, like in the TTE
						apical longitudinal view.</p>
        </sec>
        <sec>
          <title>Cranial Esophagus</title>
          <p>Short axis view: aortic arch, LVOT and pulmonary valve.</p>
          <p>Long axis view: the centrally-oriented beam allows imaging the ascending
						aorta and the LVOT; the clockwise rotation of the transducer enables imaging
						the RVOT and the PT.</p>
        </sec>
        <sec>
          <title>Novel Technology</title>
          <p>Over the past few decades, there has been a rapid and increasing development
						of novel technology and, usually, those tools need to be tested
						experimentally before they are introduced in the clinical scenario.</p>
          <p>Contrast echocardiography aims to improve the endocardial border delineation
						and, associated with the stress echocardiography, it can be employed in the
						research setting for myocardial perfusion assessment<xref ref-type="bibr" rid="B20">20</xref>. </p>
          <p>Real time 3D echocardiography, 2D and 3D STE are promising novel tools for
						the evaluation and early diagnosis of cardiac diseases<xref ref-type="bibr" rid="B21">21</xref>
						<sup>-</sup>
						<xref ref-type="bibr" rid="B24">24</xref>. The experimental validation of
						these techniques was important to support their use in the clinical setting,
						mainly considering the STE, once there is no true gold standard that might
						be applied with this purpose to humans: considering the torsional mechanics,
						the elegant study of Helle Valle et al<xref ref-type="bibr" rid="B25">25</xref>, as well as the assays of Langeland et al<xref ref-type="bibr" rid="B26">26</xref>, Amundsen et al<xref ref-type="bibr" rid="B27">27</xref>, and Seo et al<xref ref-type="bibr" rid="B28">28</xref> regarding to the 2D and 3D myocardial strain, the technique
						was validated against sonomicrometry, which demands open chest surgery and
						the insertion of crystals in the myocardium. <xref ref-type="table" rid="t04">Table 2</xref> shows the correlation and agreement between the
						gold standard and STE in these trials. However, in order to obtain reliable
						data, it is very important to proceed with the inhalation anesthesia to
						induce respiratory apnea while acquiring the images, once both the RT3DE and
						STE have their accuracy affected by the respiratory movements of chest.
						Another relevant issue concerns the suitable preset, since low temporal
						resolution, represented by the frame rate, can also interfere with the
							results.<xref ref-type="bibr" rid="B22">22</xref>. </p>
          <table-wrap id="t04">
            <label>Table 2</label>
            <caption>
              <title>Experimental validation of speckle tracking against
								sonomicrometry</title>
            </caption>
            <table rules="rows" frame="hsides">
              <thead>
                <tr style="border-bottom-style:hidden">
                  <td>
                    <bold>Study type</bold>
                  </td>
                  <td rowspan="2" align="center">
                    <bold>Parameter</bold>
                  </td>
                  <td rowspan="2" align="center">
                    <bold>Correlation</bold>
                  </td>
                </tr>
                <tr>
                  <td>
                    <bold> (Reference #)</bold>
                  </td>
                </tr>
              </thead>
              <tbody>
                <tr style="border-bottom-style:hidden">
                  <td rowspan="3">2D STE (22)</td>
                  <td align="center">Apical rotation</td>
                  <td align="center">0.92</td>
                </tr>
                <tr style="border-bottom-style:hidden">
                  <td align="center">Basal rotation</td>
                  <td align="center">0.76</td>
                </tr>
                <tr>
                  <td align="center">Twist</td>
                  <td align="center">0.94</td>
                </tr>
                <tr style="border-bottom-style:hidden">
                  <td rowspan="2">2D STE (26)</td>
                  <td align="center">Radial Strain</td>
                  <td align="center">0.72</td>
                </tr>
                <tr>
                  <td>Longitudinal Strain</td>
                  <td align="center"> 0.80</td>
                </tr>
                <tr>
                  <td>2D STE (27)</td>
                  <td align="center">Longitudinal Strain</td>
                  <td align="center">0.90</td>
                </tr>
                <tr>
                  <td>3D STE (28)</td>
                  <td align="center">Area Strain</td>
                  <td align="center">0.87</td>
                </tr>
              </tbody>
            </table>
            <table-wrap-foot>
              <fn>
                <p>2D STE: two-dimensional speckle tracking; 3D STE:
									three-dimensional speckle tracking.</p>
              </fn>
            </table-wrap-foot>
          </table-wrap>
        </sec>
      </sec>
      <sec>
        <title>Final Considerations</title>
        <p>According to the best of the authors' knowledge, the first systematic studies
					employing the echocardiography in the experimental laboratory were performed in
					the 1960's. Lategola, in 1966, through a miniaturized ultrasonic transducer
					placed inside the aorta of dogs, evaluated the stroke volume invasively, in a
					prototype that, few years later, would be clinically applied, by means of the
					transthoracic Continuous Wave Doppler<xref ref-type="bibr" rid="B29">29</xref>.
					Two years later, Christiensen and Bonte showed, in dogs, the accuracy of the
					echocardiography to detect pericardial effusion. Since then, its importance has
					been increasing in the experimental investigation scenario, seeking for clinical
						applications<xref ref-type="bibr" rid="B30">30</xref>.</p>
        <p>Unlike using other technologies, like magnetic resonance imaging, relevant
					information can be easily obtained by the echocardiography approach. According
					to the authors' experience, the method is useful to follow-up myocardial
					hypertrophy and failure<xref ref-type="bibr" rid="B31">31</xref>
					<sup>,</sup>
					<xref ref-type="bibr" rid="B32">32</xref>
					<sup>,</sup>
					<xref ref-type="bibr" rid="B12">12</xref>. </p>
        <p>The development of novel technologies, such as the ultrasound biomicroscopy,
					which uses frequencies from 30 to 100 MHz, allows the assessment of tiny
					structures - like the mice fetal heart in the uterus<xref ref-type="bibr" rid="B33">33</xref>
					<sup>-</sup>
					<xref ref-type="bibr" rid="B35">35</xref>. Certainly, it will be of extremely
					importance to shed light on issues of the cardiovascular system and to the
					development of microsurgeries in small animals. It represents a very appealing
					option in experimental research, since those animals are easily manipulated,
					have a better cost-benefit ratio, and can be maintained in small areas, enabling
					study larger samples in a shorter period of time.</p>
      </sec>
      <sec>
        <title>Author contributions</title>
        <p>Conception and design of the research: Abduch MCD, Aiello VD; Acquisition of
					data, Analysis and interpretation of the data and Critical revision of the
					manuscript for intellectual content: Abduch MCD, Assad RS, Aiello VD; Writing of
					the manuscript: Abduch MCD, Assad RS, Mathias Jr. W, Aiello VD.</p>
      </sec>
    </body>
    <back>
      <ack>
        <title>Acknowledgement</title>
        <p>We would like to thank Rodrigo Tonan for his technical assistance with the
					Figures.</p>
      </ack>
      <fn-group>
        <fn fn-type="financial-disclosure">
          <p>
            <bold>Sources of Funding</bold>
          </p>
          <p> There were no external funding sources for this study.</p>
        </fn>
        <fn fn-type="other">
          <p>
            <bold>Study Association</bold>
          </p>
          <p>This study is not associated with any post-graduation program.</p>
        </fn>
      </fn-group>
    </back>
  </sub-article>
</article>
