����JFIFXX�����    $.' ",#(7),01444'9=82<.342  2!!22222222222222222222222222222222222222222222222222����"��4�� ���,�PG"Z_�4�˷����kjز�Z�,F+��_z�,�© �����zh6�٨�ic�fu���#ډb���_�N�?��wQ���5-�~�I���8����TK<5o�Iv-�����k�_U_�����~b�M��d����Ӝ�U�Hh��?]��E�w��Q���k�{��_}qFW7HTՑ��Y��F�?_�'ϔ��_�Ջt��=||I ��6�έ"�����D���/[�k�9���Y�8ds|\���Ҿp6�Ҵ���]��.����6�z<�v��@]�i%��$j��~�g��J>��no����pM[me�i$[����s�o�ᘨ�˸ nɜG-�ĨU�ycP�3.DB�li�;��hj���x7Z^�N�h������N3u{�:j�x�힞��#M&��jL P@_���� P��&��o8������9�����@Sz6�t7#O�ߋ �s}Yf�T���lmr����Z)'N��k�۞p����w\�Tȯ?�8`�O��i{wﭹW�[�r�� ��Q4F�׊���3m&L�=��h3����z~��#�\�l :�F,j@�� ʱ�wQT����8�"kJO���6�֚l����}���R�>ډK���]��y����&����p�}b��;N�1�m�r$�|��7�>e�@B�TM*-iH��g�D�)� E�m�|�ؘbҗ�a��Ҿ����t4���o���G��*oCN�rP���Q��@z,|?W[0�����:�n,jWiE��W��$~/�hp\��?��{(�0���+�Y8rΟ�+����>S-S����VN;�}�s?.����� w�9��˟<���Mq4�Wv'��{)0�1mB��V����W[�����8�/<� �%���wT^�5���b��)iM� pg�N�&ݝ��VO~�q���u���9� ����!��J27����$O-���! �:�%H��� ـ����y�ΠM=t{!S�� oK8������t<����è:a������[�����ա�H���~��w��Qz`�po�^ ����Q��n� �,uu�C�$ ^���,������8�#��:�6��e�|~���!�3�3.�\0��q��o�4`.|� ����y�Q�`~;�d�ׯ,��O�Zw�������`73�v�܋�<���Ȏ�� ـ4k��5�K�a�u�=9Yd��$>x�A�&�� j0� ���vF��� Y�|�y��� ~�6�@c��1vOp�Ig����4��l�OD���L����� R���c���j�_�uX6��3?nk��Wy�f;^*B� ��@�~a�`��Eu������+���6�L��.ü>��}y���}_�O�6�͐�:�YrG�X��kG�����l^w���~㒶sy��Iu�!� W ��X��N�7BV��O��!X�2����wvG�R�f�T#�����t�/?���%8�^�W�aT��G�cL�M���I��(J����1~�8�?aT ���]����AS�E��(��*E}� 2��#I/�׍qz��^t�̔���b�Yz4x���t�){ OH��+(E��A&�N�������XT��o��"�XC��'���)}�J�z�p� ��~5�}�^����+�6����w��c��Q�|Lp�d�H��}�(�.|����k��c4^�"�����Z?ȕ ��a<�L�!039C� �Eu�C�F�Ew�ç ;�n?�*o���B�8�bʝ���'#Rqf���M}7����]����s2tcS{�\icTx;�\��7K���P���ʇ Z O-��~��c>"��?�������P��E��O�8��@�8��G��Q�g�a�Վ���󁶠�䧘��_%#r�>�1�z�a��eb��qcPѵ��n���#L��� =��׀t� L�7�`��V���A{�C:�g���e@�w1 Xp3�c3�ġ����p��M"'-�@n4���fG��B3�DJ�8[Jo�ߐ���gK)ƛ��$���� ���8�3�����+���� �����6�ʻ���� ���S�kI�*KZlT _`���?��K����QK�d����B`�s}�>���`��*�>��,*@J�d�oF*����弝��O}�k��s��]��y�ߘ��c1G�V���<=�7��7����6�q�PT��tXԀ�!9*4�4Tހ3XΛex�46���Y��D ����� �BdemDa����\�_l,��G�/���֌7���Y�](�xTt^%�GE�����4�}bT���ڹ�����;Y)���B�Q��u��>J/J �⮶.�XԄ��j�ݳ�+E��d ��r�5�_D�1 ��o�� �B�x�΢�#���<��W�����8���R6�@g�M�.��� dr�D��>(otU��@x=��~v���2� ӣ�d�oBd��3�eO�6�㣷�����ݜ6��6Y��Qz`��S��{���\P�~z m5{J/L��1������<�e�ͅPu�b�]�ϔ���'������f�b� Zpw��c`"��i���BD@:)ִ�:�]��hv�E�w���T�l��P���"Ju�}��وV J��G6��. J/�Qgl߭�e�����@�z�Zev2u�)]կ�����7x���s�M�-<ɯ�c��r�v�����@��$�ޮ}lk���a���'����>x��O\�ZFu>�����ck#��&:��`�$�ai�>2Δ����l���oF[h��lE�ܺ�Πk:)���`�� $[6�����9�����kOw�\|���8}������ބ:��񶐕��I�A1/�=�2[�,�!��.}gN#�u����b��� ~��݊��}34q����d�E��Lc��$��"�[q�U�硬g^��%B �z���r�pJ�ru%v\h1Y�ne`ǥ:g���pQM~�^�Xi� ��`S�:V29.�P���V�?B�k�� AEvw%�_�9C�Q����wKekPؠ�\�;Io d�{ ߞo�c1eP����\� `����E=���@K<�Y���eڼ�J���w����{av�F�'�M�@/J��+9p���|]�����Iw &`��8���&M�hg��[�{��Xj��%��Ӓ�$��(����ʹN���<>�I���RY���K2�NPlL�ɀ)��&e����B+ь����( � �JTx���_?EZ� }@ 6�U���뙢ط�z��dWI�n` D����噥�[��uV��"�G&Ú����2g�}&m��?ċ�"����Om#��������� ��{�ON��"S�X��Ne��ysQ���@Fn��Vg���dX�~nj�]J�<�K]:��FW��b�������62�=��5f����JKw��bf�X�55��~J �%^����:�-�QIE��P��v�nZum� z � ~ə ���� ���ة����;�f��\v���g�8�1��f24;�V���ǔ�)����9���1\��c��v�/'Ƞ�w�������$�4�R-��t���� e�6�/�ġ �̕Ecy�J���u�B���<�W�ַ~�w[B1L۲�-JS΂�{���΃������A��20�c#��@ 0!1@AP"#2Q`$3V�%45a6�FRUq��� ����^7ׅ,$n�������+��F�`��2X'��0vM��p�L=������5��8������u�p~���.�`r�����\���O��,ư�0oS ��_�M�����l���4�kv\JSd���x���SW�<��Ae�IX����������$I���w�:S���y���›R��9�Q[���,�5�;�@]�%���u�@ *ro�lbI �� ��+���%m:�͇ZV�����u�̉����θau<�fc�.����{�4Ա� �Q����*�Sm��8\ujqs]{kN���)qO�y�_*dJ�b�7���yQqI&9�ԌK!�M}�R�;������S�T���1���i[U�ɵz�]��U)V�S6���3$K{�ߊ<�(� E]Զ[ǼENg�����'�\?#)Dkf��J���o��v���'�%ƞ�&K�u�!��b�35LX�Ϸ��63$K�a�;�9>,R��W��3�3� d�JeTYE.Mϧ��-�o�j3+y��y^�c�������VO�9NV\nd�1 ��!͕_)a�v;����թ�M�lWR1��)El��P;��yوÏ�u 3�k�5Pr6<�⒲l�!˞*��u־�n�!�l:����UNW ��%��Chx8vL'��X�@��*��)���̮��ˍ��� ���D-M�+J�U�kvK����+�x8��cY������?�Ԡ��~3mo��|�u@[XeY�C�\Kp�x8�oC�C�&����N�~3-H���� ��MX�s�u<`���~"WL��$8ξ��3���a�)|:@�m�\���^�`�@ҷ)�5p+��6���p�%i)P M���ngc�����#0Aruz���RL+xSS?���ʮ}()#�t��mˇ!��0}}y����<�e� �-ή�Ԩ��X������ MF���ԙ~l L.3���}�V뽺�v�����멬��Nl�)�2����^�Iq��a��M��qG��T�����c3#������3U�Ǎ���}��לS�|qa��ڃ�+���-��2�f����/��bz��ڐ�� �ݼ[2�ç����k�X�2�* �Z�d���J�G����M*9W���s{��w���T��x��y,�in�O�v��]���n����P�$�JB@=4�OTI�n��e�22a\����q�d���%�$��(���:���: /*�K[PR�fr\nڙdN���F�n�$�4�[�� U�zƶ����� �mʋ���,�ao�u 3�z� �x��Kn����\[��VFmbE;�_U��&V�Gg�]L�۪&#n%�$ɯ�dG���D�TI=�%+AB�Ru#��b4�1�»x�cs�YzڙJG��f��Il��d�eF'T� iA��T���uC�$����Y��H?����[!G`}���ͪ� �纤Hv\������j�Ex�K���!���OiƸ�Yj�+u-<���'q����uN�*�r\��+�]���<�wOZ.fp�ێ��,-*)V?j-kÊ#�`�r��dV����(�ݽBk�����G�ƛk�QmUڗe��Z���f}|����8�8��a���i��3'J�����~G_�^���d�8w������ R�`(�~�.��u���l�s+g�bv���W���lGc}��u���afE~1�Ue������Z�0�8�=e�� f@/�jqEKQQ�J��oN��J���W5~M>$6�Lt�;$ʳ{���^��6�{����v6���ķܰg�V�cnn �~z�x�«�,2�u�?cE+Ș�H؎�%�Za�)���X>uW�Tz�Nyo����s���FQƤ��$��*�&�LLXL)�1�" L��eO��ɟ�9=���:t��Z���c��Ž���Y?�ӭV�wv�~,Y��r�ۗ�|�y��GaF�����C�����.�+� ���v1���fήJ�����]�S��T��B��n5sW}y�$��~z�'�c ��8 ��� ,! �p��VN�S��N�N�q��y8z˱�A��4��*��'������2n<�s���^ǧ˭P�Jޮɏ�U�G�L�J�*#��<�V��t7�8����TĜ>��i}K%,���)[��z�21z ?�N�i�n1?T�I�R#��m-�����������������1����lA�`��fT5+��ܐ�c�q՝��ʐ��,���3�f2U�եmab��#ŠdQ�y>\��)�SLY����w#��.���ʑ�f��� ,"+�w�~�N�'�c�O�3F�������N<���)j��&��,-� �љ���֊�_�zS���TǦ����w�>��?�������n��U仆�V���e�����0���$�C�d���rP �m�׈e�Xm�Vu� �L��.�bֹ��� �[Դaզ���*��\y�8�Է:�Ez\�0�Kq�C b��̘��cө���Q��=0Y��s�N��S.���3.���O�o:���#���v7�[#߫ ��5�܎�L���Er4���9n��COWlG�^��0k�%<���ZB���aB_���������'=��{i�v�l�$�uC���mƎҝ{�c㱼�y]���W�i ��ߧc��m�H� m�"�"�����;Y�ߝ�Z�Ǔ�����:S#��|}�y�,/k�Ld� TA�(�AI$+I3��;Y*���Z��}|��ӧO��d�v��..#:n��f>�>���ȶI�TX��� 8��y����"d�R�|�)0���=���n4��6ⲑ�+��r<�O�܂~zh�z����7ܓ�HH�Ga롏���nCo�>������a ���~]���R���̲c?�6(�q�;5%� |�uj�~z8R=X��I�V=�|{v�Gj\gc��q����z�؋%M�ߍ����1y��#��@f^���^�>N�����#x#۹��6�Y~�?�dfPO��{��P�4��V��u1E1J �*|���%���JN��`eWu�zk M6���q t[�� ��g�G���v��WIG��u_ft����5�j�"�Y�:T��ɐ���*�;� e5���4����q$C��2d�}���� _S�L#m�Yp��O�.�C�;��c����Hi#֩%+) �Ӎ��ƲV���SYź��g |���tj��3�8���r|���V��1#;.SQ�A[���S������#���`n�+���$��$I �P\[�@�s��(�ED�z���P��])8�G#��0B��[ى��X�II�q<��9�~[Z멜�Z�⊔IWU&A>�P~�#��dp<�?����7���c��'~���5 ��+$���lx@�M�dm��n<=e�dyX��?{�|Aef ,|n3�<~z�ƃ�uۧ�����P��Y,�ӥQ�*g�#먙R�\���;T��i,��[9Qi歉����c>]9�� ��"�c��P�� �Md?٥��If�ت�u��k��/����F��9�c*9��Ǎ:�ØF���z�n*�@|I�ށ9����N3{'��[�'ͬ�Ҳ4��#}��!�V� Fu��,�,mTIk���v C�7v���B�6k�T9��1�*l� '~��ƞF��lU��'�M ����][ΩũJ_�{�i�I�n��$���L�� j��O�dx�����kza۪��#�E��Cl����x˘�o�����V���ɞ�ljr��)�/,�߬h�L��#��^��L�ф�,íMƁe�̩�NB�L�����iL����q�}��(��q��6IçJ$�W�E$��:������=#����(�K�B����zђ <��K(�N�۫K�w��^O{!����)�H���>x�������lx�?>Պ�+�>�W���,Ly!_�D���Ō�l���Q�!�[ �S����J��1��Ɛ�Y}��b,+�Lo�x�ɓ)����=�y�oh�@�꥟/��I��ѭ=��P�y9��� �ۍYӘ�e+�p�Jnϱ?V\SO%�(�t� ���=?MR�[Ș�����d�/ ��n�l��B�7j� ��!�;ӥ�/�[-���A�>�dN�sLj ��,ɪv��=1c�.SQ�O3�U���ƀ�ܽ�E����������̻��9G�ϷD�7(�}��Ävӌ\�y�_0[w ���<΍>����a_��[0+�L��F.�޺��f�>oN�T����q;���y\��bՃ��y�jH�<|q-eɏ�_?_9+P���Hp$�����[ux�K w�Mw��N�ی'$Y2�=��q���KB��P��~������Yul:�[<����F1�2�O���5=d����]Y�sw:���Ϯ���E��j,_Q��X��z`H1,#II ��d�wr��P˂@�ZJV����y$�\y�{}��^~���[:N����ߌ�U�������O��d�����ؾe��${p>G��3c���Ė�lʌ�� ת��[��`ϱ�-W����dg�I��ig2��� ��}s ��ؤ(%#sS@���~���3�X�nRG�~\jc3�v��ӍL��M[JB�T��s3}��j�Nʖ��W����;7��ç?=X�F=-�=����q�ߚ���#���='�c��7���ڑW�I(O+=:uxq�������������e2�zi+�kuG�R��������0�&e�n���iT^J����~\jy���p'dtG��s����O��3����9* �b#Ɋ�� p������[Bws�T�>d4�ۧs���nv�n���U���_�~,�v����ƜJ1��s�� �QIz��)�(lv8M���U=�;����56��G���s#�K���MP�=��LvyGd��}�VwWBF�'�à �?MH�U�g2�� ����!�p�7Q��j��ڴ����=��j�u��� Jn�A s���uM������e��Ɔ�Ҕ�!)'��8Ϣ�ٔ��ޝ(��Vp���צ֖d=�IC�J�Ǡ{q������kԭ�߸���i��@K����u�|�p=..�*+����x�����z[Aqġ#s2a�Ɗ���RR�)*HRsi�~�a &f��M��P����-K�L@��Z��Xy�'x�{}��Zm+���:�)�) IJ�-i�u���� ���ܒH��'�L(7�y�GӜq���� j��� 6ߌg1�g�o���,kر���tY�?W,���p���e���f�OQS��!K�۟cҒA�|ս�j�>��=⬒��˧L[�� �߿2JaB~R��u�:��Q�] �0H~���]�7��Ƽ�I���(}��cq '�ήET���q�?f�ab���ӥvr� �)o��-Q��_'����ᴎo��K������;��V���o��%���~OK ����*��b�f:���-ťIR��`B�5!RB@���ï�� �u �̯e\�_U�_������� g�ES��3�������QT��a����x����U<~�c?�*�#]�MW,[8O�a�x��]�1bC|踤�P��lw5V%�)�{t�<��d��5���0i�XSU��m:��Z�┵�i�"��1�^B�-��P�hJ��&)O��*�D��c�W��vM��)����}���P��ܗ-q����\mmζZ-l@�}��a��E�6��F�@��&Sg@���ݚ�M����� ȹ 4����#p�\H����dYDo�H���"��\��..R�B�H�z_�/5˘����6��KhJR��P�mƶi�m���3�,#c�co��q�a)*Pt����R�m�k�7x�D�E�\Y�閣_X�<���~�)���c[[�BP����6�Yq���S��0����%_����;��Àv�~�| VS؇ ��'O0��F0��\���U�-�d@�����7�SJ*z��3n��y��P����O���������m�~�P�3|Y��ʉr#�C�<�G~�.,! ���bqx���h~0=��!ǫ�jy����l�O,�[B��~��|9��ٱ����Xly�#�i�B��g%�S��������tˋ���e���ې��\[d�t)��.+u�|1 ������#�~Oj����hS�%��i.�~X���I�H�m��0n���c�1uE�q��cF�RF�o���7� �O�ꮧ� ���ۛ{��ʛi5�rw?׌#Qn�TW��~?y$��m\�\o����%W� ?=>S�N@�� �Ʈ���R����N�)�r"C�:��:����� �����#��qb��Y�. �6[��2K����2u�Ǧ�HYR��Q�MV��� �G�$��Q+.>�����nNH��q�^��� ����q��mM��V��D�+�-�#*�U�̒ ���p욳��u:�������IB���m���PV@O���r[b= �� ��1U�E��_Nm�yKbN�O���U�}�the�`�|6֮P>�\2�P�V���I�D�i�P�O;�9�r�mAHG�W�S]��J*�_�G��+kP�2����Ka�Z���H�'K�x�W�MZ%�O�YD�Rc+o��?�q��Ghm��d�S�oh�\�D�|:W������UA�Qc yT�q������~^�H��/��#p�CZ���T�I�1�ӏT����4��"�ČZ�����}��`w�#�*,ʹ�� ��0�i��課�Om�*�da��^gJ݅{���l�e9uF#T�ֲ��̲�ٞC"�q���ߍ ոޑ�o#�XZTp����@ o�8��(jd��xw�]�,f���`~�|,s��^����f�1���t��|��m�򸄭/ctr��5s��7�9Q�4�H1꠲BB@l9@���C�����+�wp�xu�£Yc�9��?`@#�o�mH�s2��)�=��2�.�l����jg�9$�Y�S�%*L������R�Y������7Z���,*=�䷘$�������arm�o�ϰ���UW.|�r�uf����IGw�t����Zwo��~5 ��YյhO+=8fF�)�W�7�L9lM�̘·Y���֘YLf�큹�pRF���99.A �"wz��=E\Z���'a� 2��Ǚ�#;�'}�G���*��l��^"q��+2FQ� hj��kŦ��${���ޮ-�T�٭cf�|�3#~�RJ����t��$b�(R��(����r���dx� >U b�&9,>���%E\� Ά�e�$��'�q't��*�א���ެ�b��-|d���SB�O�O��$�R+�H�)�܎�K��1m`;�J�2�Y~9��O�g8=vqD`K[�F)k�[���1m޼c��n���]s�k�z$@��)!I �x՝"v��9=�ZA=`Ɠi �:�E��)`7��vI��}d�YI�_ �o�:ob���o ���3Q��&D&�2=�� �Ά��;>�h����y.*ⅥS������Ӭ�+q&����j|UƧ����}���J0��WW< ۋS�)jQR�j���Ư��rN)�Gű�4Ѷ(�S)Ǣ�8��i��W52���No˓� ۍ%�5brOn�L�;�n��\G����=�^U�dI���8$�&���h��'���+�(������cȁ߫k�l��S^���cƗjԌE�ꭔ��gF���Ȓ��@���}O���*;e�v�WV���YJ\�]X'5��ղ�k�F��b 6R�o՜m��i N�i����>J����?��lPm�U��}>_Z&�KK��q�r��I�D�Չ~�q�3fL�:S�e>���E���-G���{L�6p�e,8��������QI��h��a�Xa��U�A'���ʂ���s�+טIjP�-��y�8ۈZ?J$��W�P� ��R�s�]��|�l(�ԓ��sƊi��o(��S0��Y� 8�T97.�����WiL��c�~�dxc�E|�2!�X�K�Ƙਫ਼�$((�6�~|d9u+�qd�^3�89��Y�6L�.I�����?���iI�q���9�)O/뚅����O���X��X�V��ZF[�یgQ�L��K1���RҖr@v�#��X�l��F���Нy�S�8�7�kF!A��sM���^rkp�jP�DyS$N���q��nxҍ!U�f�!eh�i�2�m���`�Y�I�9r�6� �TF���C}/�y�^���Η���5d�'��9A-��J��>{�_l+�`��A���[�'��յ�ϛ#w:݅�%��X�}�&�PSt�Q�"�-��\縵�/����$Ɨh�Xb�*�y��BS����;W�ջ_mc�����vt?2}1�;qS�d�d~u:2k5�2�R�~�z+|HE!)�Ǟl��7`��0�<�,�2*���Hl-��x�^����'_TV�gZA�'j� ^�2Ϊ��N7t�����?w�� �x1��f��Iz�C-Ȗ��K�^q�;���-W�DvT�7��8�Z�������� hK�(P:��Q- �8�n�Z���܃e貾�<�1�YT<�,�����"�6{/ �?�͟��|1�:�#g��W�>$����d��J��d�B��=��jf[��%rE^��il:��B���x���Sּ�1հ��,�=��*�7 fcG��#q� �eh?��2�7�����,�!7x��6�n�LC�4x��},Geǝ�tC.��vS �F�43��zz\��;QYC,6����~;RYS/6���|2���5���v��T��i����������mlv��������&� �nRh^ejR�LG�f���? �ۉҬܦƩ��|��Ȱ����>3����!v��i�ʯ�>�v��オ�X3e���_1z�Kȗ\<������!�8���V��]��?b�k41�Re��T�q��mz��TiOʦ�Z��Xq���L������q"+���2ۨ��8}�&N7XU7Ap�d�X��~�׿��&4e�o�F��� �H����O���č�c�� 懴�6���͉��+)��v;j��ݷ�� �UV�� i��� j���Y9GdÒJ1��詞�����V?h��l����l�cGs�ځ�������y�Ac�����\V3�? �� ܙg�>qH�S,�E�W�[�㺨�uch�⍸�O�}���a��>�q�6�n6����N6�q������N ! 1AQaq�0@����"2BRb�#Pr���3C`��Scst���$4D���%Td�� ?���N����a��3��m���C���w��������xA�m�q�m���m������$����4n淿t'��C"w��zU=D�\R+w�p+Y�T�&�պ@��ƃ��3ޯ?�Aﶂ��aŘ���@-�����Q�=���9D��ռ�ѻ@��M�V��P��܅�G5�f�Y<�u=,EC)�<�Fy'�"�&�չ�X~f��l�KԆV��?�� �W�N����=(� �;���{�r����ٌ�Y���h{�١������jW����P���Tc�����X�K�r��}���w�R��%��?���E��m�� �Y�q|����\lEE4���r���}�lsI�Y������f�$�=�d�yO����p�����yBj8jU�o�/�S��?�U��*������ˍ�0������u�q�m [�?f����a�� )Q�>����6#������� ?����0UQ����,IX���(6ڵ[�DI�MNލ�c&���υ�j\��X�R|,4��� j������T�hA�e��^���d���b<����n�� �즇�=!���3�^�`j�h�ȓr��jẕ�c�,ٞX����-����a�ﶔ���#�$��]w�O��Ӫ�1y%��L�Y<�wg#�ǝ�̗`�x�xa�t�w��»1���o7o5��>�m뭛C���Uƃߜ}�C���y1Xνm�F8�jI���]����H���ۺиE@I�i;r�8ӭ����V�F�Շ| ��&?�3|x�B�MuS�Ge�=Ӕ�#BE5G�����Y!z��_e��q�р/W>|-�Ci߇�t�1ޯќd�R3�u��g�=0 5��[?�#͏��q�cf���H��{ ?u�=?�?ǯ���}Z��z���hmΔ�BFTW�����<�q�(v� ��!��z���iW]*�J�V�z��gX֧A�q�&��/w���u�gYӘa���; �i=����g:��?2�dž6�ى�k�4�>�Pxs����}������G�9��3 ���)gG�R<>r h�$��'nc�h�P��Bj��J�ҧH� -��N1���N��?��~��}-q!=��_2hc�M��l�vY%UE�@|�v����M2�.Y[|y�"Eï��K�ZF,�ɯ?,q�?v�M 80jx�"�;�9vk�����+ ֧�� �ȺU��?�%�vcV��mA�6��Qg^M����A}�3�nl� QRN�l8�kkn�'�����(��M�7m9و�q���%ޟ���*h$Zk"��$�9��: �?U8�Sl��,,|ɒ��xH(ѷ����Gn�/Q�4�P��G�%��Ա8�N��!� �&�7�;���eKM7�4��9R/%����l�c>�x;������>��C�:�����t��h?aKX�bhe�ᜋ^�$�Iհ �hr7%F$�E��Fd���t��5���+�(M6�t����Ü�UU|zW�=a�Ts�Tg������dqP�Q����b'�m���1{|Y����X�N��b �P~��F^F:����k6�"�j!�� �I�r�`��1&�-$�Bevk:y���#yw��I0��x��=D�4��tU���P�ZH��ڠ底taP��6����b>�xa����Q�#� WeF��ŮNj�p�J* mQ�N����*I�-*�ȩ�F�g�3 �5��V�ʊ�ɮ�a��5F���O@{���NX��?����H�]3��1�Ri_u��������ѕ�� ����0��� F��~��:60�p�͈�S��qX#a�5>���`�o&+�<2�D����: �������ڝ�$�nP���*)�N�|y�Ej�F�5ټ�e���ihy�Z �>���k�bH�a�v��h�-#���!�Po=@k̆IEN��@��}Ll?j�O������߭�ʞ���Q|A07x���wt!xf���I2?Z��<ץ�T���cU�j��]��陎Ltl �}5�ϓ��$�,��O�mˊ�;�@O��jE��j(�ا,��LX���LO���Ц�90�O �.����a��nA���7������j4 ��W��_ٓ���zW�jcB������y՗+EM�)d���N�g6�y1_x��p�$Lv:��9�"z��p���ʙ$��^��JԼ*�ϭ����o���=x�Lj�6�J��u82�A�H�3$�ٕ@�=Vv�]�'�qEz�;I˼��)��=��ɯ���x �/�W(V���p�����$ �m�������u�����񶤑Oqˎ�T����r��㠚x�sr�GC��byp�G��1ߠ�w e�8�$⿄����/�M{*}��W�]˷.�CK\�ުx���/$�WPw���r� |i���&�}�{�X� �>��$-��l���?-z���g����lΆ���(F���h�vS*���b���߲ڡn,|)mrH[���a�3�ר�[1��3o_�U�3�TC�$��(�=�)0�kgP���� ��u�^=��4 �WYCҸ:��vQ�ר�X�à��tk�m,�t*��^�,�}D*� �"(�I��9R����>`�`��[~Q]�#af��i6l��8���6�:,s�s�N6�j"�A4���IuQ��6E,�GnH��zS�HO�uk�5$�I�4��ؤ�Q9�@��C����wp�BGv[]�u�Ov���0I4���\��y�����Q�Ѹ��~>Z��8�T��a��q�ޣ;z��a���/��S��I:�ܫ_�|������>=Z����8:�S��U�I�J��"IY���8%b8���H��:�QO�6�;7�I�S��J��ҌAά3��>c���E+&jf$eC+�z�;��V����� �r���ʺ������my�e���aQ�f&��6�ND��.:��NT�vm�<- u���ǝ\MvZY�N�NT��-A�>jr!S��n�O 1�3�Ns�%�3D@���`������ܟ 1�^c<���� �a�ɽ�̲�Xë#�w�|y�cW�=�9I*H8�p�^(4���՗�k��arOcW�tO�\�ƍR��8����'�K���I�Q�����?5�>[�}��yU�ײ -h��=��% q�ThG�2�)���"ו3]�!kB��*p�FDl�A���,�eEi�H�f�Ps�����5�H:�Փ~�H�0Dت�D�I����h�F3�������c��2���E��9�H��5�zԑ�ʚ�i�X�=:m�xg�hd(�v����׊�9iS��O��d@0ڽ���:�p�5�h-��t�&���X�q�ӕ,��ie�|���7A�2���O%P��E��htj��Y1��w�Ѓ!����  ���� ࢽ��My�7�\�a�@�ţ�J �4�Ȼ�F�@o�̒?4�wx��)��]�P��~�����u�����5�����7X ��9��^ܩ�U;Iꭆ 5 �������eK2�7(�{|��Y׎ �V��\"���Z�1� Z�����}��(�Ǝ"�1S���_�vE30>���p;� ΝD��%x�W�?W?v����o�^V�i�d��r[��/&>�~`�9Wh��y�;���R��� ;;ɮT��?����r$�g1�K����A��C��c��K��l:�'��3 c�ﳯ*"t8�~l��)���m��+U,z��`(�>yJ�?����h>��]��v��ЍG*�{`��;y]��I�T� ;c��NU�fo¾h���/$���|NS���1�S�"�H��V���T���4��uhǜ�]�v;���5�͠x��'C\�SBpl���h}�N����� A�Bx���%��ޭ�l��/����T��w�ʽ]D�=����K���ž�r㻠l4�S�O?=�k �M:� ��c�C�a�#ha���)�ѐxc�s���gP�iG��{+���x���Q���I= �� z��ԫ+ �8"�k�ñ�j=|����c ��y��CF��/��*9ж�h{ �?4�o� ��k�m�Q�N�x��;�Y��4膚�a�w?�6�>e]�����Q�r�:����g�,i"�����ԩA�*M�<�G��b�if��l^M��5� �Ҩ�{����6J��ZJ�����P�*�����Y���ݛu�_4�9�I8�7���������,^ToR���m4�H��?�N�S�ѕw��/S��甍�@�9H�S�T��t�ƻ���ʒU��*{Xs�@����f�����֒Li�K{H�w^���������Ϥm�tq���s� ���ք��f:��o~s��g�r��ט� �S�ѱC�e]�x���a��) ���(b-$(�j>�7q�B?ӕ�F��hV25r[7 Y� }L�R��}����*sg+��x�r�2�U=�*'WS��ZDW]�WǞ�<��叓���{�$�9Ou4��y�90-�1�'*D`�c�^o?(�9��u���ݐ��'PI&� f�Jݮ�������:wS����jfP1F:X �H�9dԯ���˝[�_54 �}*;@�ܨ�� ð�yn�T���?�ןd�#���4rG�ͨ��H�1�|-#���Mr�S3��G�3�����)�.᧏3v�z֑��r����$G"�`j �1t��x0<Ɔ�Wh6�y�6��,œ�Ga��gA����y��b��)��h�D��ß�_�m��ü �gG;��e�v��ݝ�nQ� ��C����-�*��o���y�a��M��I�>�<���]obD��"�:���G�A��-\%LT�8���c�)��+y76���o�Q�#*{�(F�⽕�y����=���rW�\p���۩�c���A���^e6��K������ʐ�cVf5$�'->���ՉN"���F�"�UQ@�f��Gb~��#�&�M=��8�ט�JNu9��D��[̤�s�o�~������ G��9T�tW^g5y$b��Y'��س�Ǵ�=��U-2 #�MC�t(�i� �lj�@Q 5�̣i�*�O����s�x�K�f��}\��M{E�V�{�υ��Ƈ�����);�H����I��fe�Lȣr�2��>��W�I�Ȃ6������i��k�� �5�YOxȺ����>��Y�f5'��|��H+��98pj�n�.O�y�������jY��~��i�w'������l�;�s�2��Y��:'lg�ꥴ)o#'Sa�a�K��Z� �m��}�`169�n���"���x��I ��*+� }F<��cГ���F�P�������ֹ*�PqX�x۩��,� ��N�� �4<-����%����:��7����W���u�`����� $�?�I��&����o��o��`v�>��P��"��l���4��5'�Z�gE���8���?��[�X�7(��.Q�-��*���ތL@̲����v��.5���[��=�t\+�CNܛ��,g�SQnH����}*F�G16���&:�t��4ُ"A��̣��$�b �|����#rs��a�����T�� ]�<�j��BS�('$�ɻ� �wP;�/�n��?�ݜ��x�F��yUn�~mL*-�������Xf�wd^�a�}��f�,=t�׵i�.2/wpN�Ep8�OР���•��R�FJ� 55TZ��T �ɭ�<��]��/�0�r�@�f��V��V����Nz�G��^���7hZi����k��3�,kN�e|�vg�1{9]_i��X5y7� 8e]�U����'�-2,���e"����]ot�I��Y_��n�(JҼ��1�O ]bXc���Nu�No��pS���Q_���_�?i�~�x h5d'�(qw52] ��'ޤ�q��o1�R!���`ywy�A4u���h<קy���\[~�4�\ X�Wt/� 6�����n�F�a8��f���z �3$�t(���q��q�x��^�XWeN'p<-v�!�{�(>ӽDP7��ո0�y)�e$ٕv�Ih'Q�EA�m*�H��RI��=:��� ���4牢) �%_iN�ݧ�l]� �Nt���G��H�L��� ɱ�g<���1V�,�J~�ٹ�"K��Q�� 9�HS�9�?@��k����r�;we݁�]I�!{ �@�G�[�"��`���J:�n]�{�cA�E����V��ʆ���#��U9�6����j�#Y�m\��q�e4h�B�7��C�������d<�?J����1g:ٳ���=Y���D�p�ц� ׈ǔ��1�]26؜oS�'��9�V�FVu�P�h�9�xc�oq�X��p�o�5��Ա5$�9W�V(�[Ak�aY錎qf;�'�[�|���b�6�Ck��)��#a#a˙��8���=äh�4��2��C��4tm^ �n'c���]GQ$[Wҿ��i���vN�{Fu ��1�gx��1┷���N�m��{j-,��x�� Ūm�ЧS�[�s���Gna���䑴�� x�p 8<������97�Q���ϴ�v�aϚG��Rt�Һ׈�f^\r��WH�JU�7Z���y)�vg=����n��4�_)y��D'y�6�]�c�5̪�\� �PF�k����&�c;��cq�$~T�7j ���nç]�<�g ":�to�t}�159�<�/�8������m�b�K#g'I'.W�����6��I/��>v��\�MN��g���m�A�yQL�4u�Lj�j9��#44�t��l^�}L����n��R��!��t��±]��r��h6ٍ>�yҏ�N��fU�� ���� Fm@�8}�/u��jb9������he:A�y�ծw��GpΧh�5����l}�3p468��)U��d��c����;Us/�֔�YX�1�O2��uq�s��`hwg�r~�{ R��mhN��؎*q 42�*th��>�#���E����#��Hv�O����q�}�����6�e��\�,Wk�#���X��b>��p}�դ��3���T5��†��6��[��@�P�y*n��|'f�֧>�lư΂�̺����SU�'*�q�p�_S�����M�� '��c�6�����m�� ySʨ;M��r���Ƌ�m�Kxo,���Gm�P��A�G�:��i��w�9�}M(�^�V��$ǒ�ѽ�9���|���� �a����J�SQ�a���r�B;����}���ٻ֢�2�%U���c�#�g���N�a�ݕ�'�v�[�OY'��3L�3�;,p�]@�S��{ls��X�'���c�jw�k'a�.��}�}&�� �dP�*�bK=ɍ!����;3n�gΊU�ߴmt�'*{,=SzfD� A��ko~�G�aoq�_mi}#�m�������P�Xhύ����mxǍ�΂���巿zf��Q���c���|kc�����?���W��Y�$���_Lv����l߶��c���`?����l�j�ݲˏ!V��6����U�Ђ(A���4y)H���p�Z_�x��>���e��R��$�/�`^'3qˏ�-&Q�=?��CFVR �D�fV�9��{�8g�������n�h�(P"��6�[�D���< E�����~0<@�`�G�6����Hг�cc�� �c�K.5��D��d�B���`?�XQ��2��ٿyqo&+�1^� DW�0�ꊩ���G�#��Q�nL3��c���������/��x ��1�1[y�x�პCW��C�c�UĨ80�m�e�4.{�m��u���I=��f�����0QRls9���f���������9���~f�����Ǩ��a�"@�8���ȁ�Q����#c�ic������G��$���G���r/$W�(��W���V�"��m�7�[m�A�m����bo��D� j����۳� l���^�k�h׽����� ��#� iXn�v��eT�k�a�^Y�4�BN��ĕ��0 !01@Q"2AaPq3BR������?���@4�Q�����T3,���㺠�W�[=JK�Ϟ���2�r^7��vc�:�9 �E�ߴ�w�S#d���Ix��u��:��Hp��9E!�� V 2;73|F��9Y���*ʬ�F��D����u&���y؟��^EA��A��(ɩ���^��GV:ݜDy�`��Jr29ܾ�㝉��[���E;Fzx��YG��U�e�Y�C���� ����v-tx����I�sם�Ę�q��Eb�+P\ :>�i�C'�;�����k|z�رn�y]�#ǿb��Q��������w�����(�r|ӹs��[�D��2v-%��@;�8<a���[\o[ϧw��I!��*0�krs)�[�J9^��ʜ��p1)� "��/_>��o��<1����A�E�y^�C��`�x1'ܣn�p��s`l���fQ��):�l����b>�Me�jH^?�kl3(�z:���1ŠK&?Q�~�{�ٺ�h�y���/�[��V�|6��}�KbX����mn[-��7�5q�94�������dm���c^���h� X��5��<�eޘ>G���-�}�دB�ޟ� ��|�rt�M��V+�]�c?�-#ڛ��^ǂ}���Lkr���O��u�>�-D�ry� D?:ޞ�U��ǜ�7�V��?瓮�"�#���r��չģVR;�n���/_� ؉v�ݶe5d�b9��/O��009�G���5n�W����JpA�*�r9�>�1��.[t���s�F���nQ� V 77R�]�ɫ8����_0<՜�IF�u(v��4��F�k�3��E)��N:��yڮe��P�`�1}�$WS��J�SQ�N�j�ٺ��޵�#l���ј(�5=��5�lǏmoW�v-�1����v,W�mn��߀$x�<����v�j(����c]��@#��1������Ǔ���o'��u+����;G�#�޸��v-lη��/(`i⣍Pm^���ԯ̾9Z��F��������n��1��� ��]�[��)�'������:�֪�W��FC����� �B9،!?���]��V��A�Վ�M��b�w��G F>_DȬ0¤�#�QR�[V��kz���m�w�"��9ZG�7'[��=�Q����j8R?�zf�\a�=��O�U����*oB�A�|G���2�54 �p��.w7� �� ��&������ξxGHp� B%��$g�����t�Џ򤵍z���HN�u�Я�-�'4��0��;_��3 !01"@AQa2Pq#3BR������?��ʩca��en��^��8���<�u#��m*08r��y�N"�<�Ѳ0��@\�p��� �����Kv�D��J8�Fҽ� �f�Y��-m�ybX�NP����}�!*8t(�OqѢ��Q�wW�K��ZD��Δ^e��!� ��B�K��p~�����e*l}z#9ң�k���q#�Ft�o��S�R����-�w�!�S���Ӥß|M�l޶V��!eˈ�8Y���c�ЮM2��tk���� ������J�fS����Ö*i/2�����n]�k�\���|4yX�8��U�P.���Ы[���l��@"�t�<������5�lF���vU�����W��W��;�b�cД^6[#7@vU�xgZv��F�6��Q,K�v��� �+Ъ��n��Ǣ��Ft���8��0��c�@�!�Zq s�v�t�;#](B��-�nῃ~���3g������5�J�%���O������n�kB�ĺ�.r��+���#�N$?�q�/�s�6��p��a����a��J/��M�8��6�ܰ"�*������ɗud"\w���aT(����[��F��U՛����RT�b���n�*��6���O��SJ�.�ij<�v�MT��R\c��5l�sZB>F��<7�;EA��{��E���Ö��1U/�#��d1�a�n.1ě����0�ʾR�h��|�R��Ao�3�m3 ��%�� ���28Q� ��y��φ���H�To�7�lW>����#i`�q���c����a��� �m,B�-j����݋�'mR1Ήt�>��V��p���s�0IbI�C.���1R�ea�����]H�6����������4B>��o��](��$B���m�����a�!=��?�B� K�Ǿ+�Ծ"�n���K��*��+��[T#�{E�J�S����Q�����s�5�:�U�\wĐ�f�3����܆&�)����I���Ԇw��E T�lrTf6Q|R�h:��[K�� �z��c֧�G�C��%\��_�a�84��HcO�bi��ؖV��7H �)*ģK~Xhչ0��4?�0��� �E<���}3���#���u�?�� ��|g�S�6ꊤ�|�I#Hڛ� �ա��w�X��9��7���Ŀ%�SL��y6č��|�F�a 8���b��$�sק�h���b9RAu7�˨p�Č�_\*w��묦��F ����4D~�f����|(�"m���NK��i�S�>�$d7SlA��/�²����SL��|6N�}���S�˯���g��]6��; �#�.��<���q'Q�1|KQ$�����񛩶"�$r�b:���N8�w@��8$�� �AjfG|~�9F ���Y��ʺ��Bwؒ������M:I岎�G��`s�YV5����6��A �b:�W���G�q%l�����F��H���7�������Fsv7��k�� 403WebShell
403Webshell
Server IP : 172.67.215.126  /  Your IP : 172.69.130.107
Web Server : Apache/2.4.52 (Ubuntu)
System : Linux ip-172-31-19-221 6.8.0-1029-aws #31~22.04.1-Ubuntu SMP Thu Apr 24 21:16:18 UTC 2025 x86_64
User : www-data ( 33)
PHP Version : 8.1.28
Disable Function : NONE
MySQL : OFF  |  cURL : ON  |  WGET : ON  |  Perl : ON  |  Python : OFF  |  Sudo : ON  |  Pkexec : ON
Directory :  /efsdata/scitechnol.com/httpdocs/submission/uploads/

Upload File :
current_dir [ Writeable ] document_root [ Writeable ]

 

Command :


[ Back ]     

Current File : /efsdata/scitechnol.com/httpdocs/submission/uploads/1737-1-Manuscript-2014-03-21.doc
��ࡱ�>��	),����&'(�����������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������[�	���$bjbj����
;6ΐΐ��+�������vv������������8	 ��!��]E�"��$�$�$�$�%�%�%@DBDBDBDBDBDBDwG�JFBD���%�%�%�%�%BD���$�$�E�1�1�1�%L	��$��$�?��1�%@D�1�1V:8@�8�$�����iG�E���.`z8�?-E0]E�8x_J\0�_J�8�88_J�69`�%�%�1�%�%�%�%�%BDBD�1�%�%�%]E�%�%�%�%��������������������������������������������������������������������_J�%�%�%�%�%�%�%�%�%v
�:
,Title

Fluorescence polarization assay for the diagnosis of anti-Brucella abortus antibodies in cattle serum: adaptation for its use in microplates and comparison with conventional agglutination tests

Authors:
S.E. Guti�rrez1,3,* , C.M. L�tzelschwab1,2, A.G. D�az2,3 and S.M.Estein2,3

1Laboratorio de Virolog�a y 2Laboratorio de Inmunolog�a, Departamento de Sanidad Animal y Medicina Preventiva, Facultad de Ciencias Veterinarias, Universidad Nacional del Centro de la provincia de Buenos Aires. Centro de Investigaciones en Veterinaria de Tandil (CIVETAN), Consejo Nacional de Investigaciones Cient�ficas y T�cnicas (CONICET), Pinto 399, Tandil, CP 7000, Argentina.
3CONICET

*Corresponding author
Laboratorio de Virolog�a, departamento SAMP, CIVETAN-CONICET, Facultad de Ciencias Veterinarias, UNCPBA. Calle Pinto 399, Tandil, CP 7000, Argentina
Phone: 54-249-4439850 int 255.
Fax: 54-249-4439850.
e-mail: segutier@vet.unicen.edu.ar 
Abstract

Bovine brucellosis, which is endemic in Argentina, is controlled by vaccination and culling of infected cattle. Conventional agglutination tests and primary binding assays like ELISA and fluorescence polarization assay (FPA) are used for the identification of infected cattle. The FPA has many advantages over the agglutination tests, however most accredited laboratories still use the conventional agglutination tests. FPA has been extensively evaluated in its original format of 10mm x 75mm glass tubes, while there are little reports on its performance in the 96-well microplate format. The aim of the present study was to set the conditions for the use of a commercially available antigen (the O-polysaccharide from B. abortus 1119-3 conjugated with fluorescein isothiocyanate) for the FPA assay in a 96-well microplate format, and to compare its diagnostic performance with the conventional agglutination tests currently used in Argentina. Serum samples were obtained from 149 cows and 20 bulls belonging to free and infected herds from different regions of Argentina. Two dilutions of serum and antigen were assayed and the fluorescence polarization was detected with a Beckman DTX 880 multimode reader. Results: The FPA conducted using antigen and serum at 1:100 dilution was as efficient as the test using antigen at 1:20 and serum at 1:10. Eight percent sera from an endemic region gave false positive reaction in the agglutination tests compared to FPA, while 8% of samples giving suspected result in agglutination test resulted negative in FPA. Due to its high testing capacity, economy of antigen, lack of false positive reactions, and avoidance of re-testing of cattle with suspected result in agglutination tests, the FPA conducted in microplates is a suitable test to be implemented in the control and eradication of brucellosis in Argentina.

Keywords
brucellosis, fluorescence polarization, agglutination, microplate, serology

Abbreviations
FPA: fluorescence polarization assay
BPAT: buffered plate antigen test
SAT: serum agglutination test
b�-ME: b� mercaptoethanol
O-PS: O-polysaccharide

Introduction

Bovine brucellosis is one of the most common bacterial zoonosis in the world and is responsible for important economic losses in cattle. The infection in cattle is mainly due to Brucella abortus and is characterized by the death and expulsion of the foetus between the fifth and eighth months of gestation. Diagnosis of brucellosis is complicated due to its variable incubation time and the absence of clinical signs other than abortion. Serological techniques are the mainstay of diagnosis and mass testing programmes.
Brucellosis is endemic in Argentina. A national survey in 2004 showed that 12.4% of the beef farms were seropositive to Brucella, and that the apparent prevalence was 2.15%  ADDIN EN.CITE <EndNote><Cite><Author>Aznar</Author><Year>2012</Year><RecNum>45</RecNum><record><rec-number>45</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">45</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Aznar, M. N.</author><author>Samartino, L. E.</author><author>Humblet, M. F.</author><author>Saegerman, C.</author></authors></contributors><auth-address>INTA, CICVyA, Instituto de Patobiologia, Unidad de Epidemiologia, Buenos Aires, Argentina; Faculty of Veterinary Medicine, Department of Infectious and Parasitic Diseases, Research Unit of Epidemiology and Risk Analysis Applied to Veterinary Sciences (UREAR-ULg), University of Liege, Liege, Belgium.</auth-address><titles><title>Bovine Brucellosis in Argentina and Bordering Countries: Update</title><secondary-title>Transbound Emerg Dis</secondary-title></titles><periodical><full-title>Transbound Emerg Dis</full-title></periodical><edition>2012/10/11</edition><dates><year>2012</year><pub-dates><date>Oct 10</date></pub-dates></dates><isbn>1865-1682 (Electronic)</isbn><accession-num>23046031</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=23046031</url></related-urls></urls><electronic-resource-num>10.1111/tbed.12018</electronic-resource-num><language>Eng</language></record></Cite></EndNote>[1]. The National Program of Control and Eradication of Bovine Brucellosis in this country involves the vaccination of female calves between 3 and 8 months of age with B. abortus S19 and culling of infected cattle. In Argentina, the identification of infected cattle is carried out by serological tests: buffered plate antigen test (BPAT), indirect ELISA and fluorescence polarization assay (FPA) are used as screening tests, while serum agglutination test with b�-mercaptoethanol (SAT/b�-ME) and competitive ELISA tests are used as complementary. At present, complement fixation test is considered the confirmatory test. All tests detect antibodies to the O-chain of smooth lipopolysaccharide induced by vaccination or by Brucella field strains  ADDIN EN.CITE  ADDIN EN.CITE.DATA [2,3]. Occasionally, false positive reactions were found in healthy adult bovine induced by cross reaction antibodies stimulated by the administration of vaccines composed of Gram negative bacteria, but the cross reaction between these bacteria and B. abortus is dependent on the test used. Thus, extensive cross reaction was observed with the indirect ELISA with much less reactivity in the FPA and the competitive ELISA  ADDIN EN.CITE  ADDIN EN.CITE.DATA [4].
The FPA is a primary binding homogeneous immunoassay used for the detection of anti-B. abortus O-PS (O-polysaccharide) antibodies. The method is based on the principle that the rate of rotation of a small labeled antigen molecule in solution is altered if antibody is bound to it, and this change in rotation can be measured. Nielsen et al  ADDIN EN.CITE <EndNote><Cite><Author>Nielsen</Author><Year>1996</Year><RecNum>1</RecNum><record><rec-number>1</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">1</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Nielsen, K.</author><author>Gall, D.</author><author>Jolley, M.</author><author>Leishman, G.</author><author>Balsevicius, S.</author><author>Smith, P.</author><author>Nicoletti, P.</author><author>Thomas, F.</author></authors></contributors><auth-address>Agriculture and Agri-Food Canada, Animal Diseases Research Institute, Nepean, Ont., Canada.</auth-address><titles><title>A homogeneous fluorescence polarization assay for detection of antibody to Brucella abortus</title><secondary-title>J Immunol Methods</secondary-title></titles><periodical><full-title>J Immunol Methods</full-title></periodical><pages>161-8</pages><volume>195</volume><number>1-2</number><edition>1996/09/09</edition><keywords><keyword>Animals</keyword><keyword>Antibodies, Bacterial/*blood</keyword><keyword>Brucella abortus/*immunology</keyword><keyword>Cattle</keyword><keyword>*Fluorescence Polarization Immunoassay</keyword></keywords><dates><year>1996</year><pub-dates><date>Sep 9</date></pub-dates></dates><isbn>0022-1759 (Print)&#xD;0022-1759 (Linking)</isbn><accession-num>8814332</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=8814332</url></related-urls></urls><language>eng</language></record></Cite></EndNote>[5] developed a FPA using the O-PS from B. abortus conjugated with fluorescein isothiocyanate as a tracer for detection of antibody to B. abortus in bovine serum. The main advantages of the FPA are its rapidness, simplicity, specificity, objectivity of results, and the ability to early distinguish vaccinated animals from those infected with Brucella spp.  ADDIN EN.CITE  ADDIN EN.CITE.DATA [6,7]. Diagnostic performance of the FPA was determined in well characterized samples from Canada, resulting in 99% sensitivity and 99.96% specificity. Due to these properties, the World Organization for Animal Health (OIE) has recommended the FPA assay as a compulsory test for the international trade of cattle  ADDIN EN.CITE <EndNote><Cite><Author>OIE</Author><Year>2009</Year><RecNum>73</RecNum><record><rec-number>73</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">73</key></foreign-keys><ref-type name="Book Section">5</ref-type><contributors><authors><author>OIE</author></authors></contributors><titles><title>Chapter 2.4.3.:Bovine brucellosis</title><secondary-title>Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. </secondary-title></titles><dates><year>2009</year></dates><publisher>Office International des Epizooties, Paris.</publisher><urls></urls></record></Cite></EndNote>[8]. The test has been extensively used with sera from different species of animals and in areas of various incidences of brucellosis. It was originally developed to be conducted in 10mm x75mm glass tubes in a single cell analyzer. The availability of microplate readers favors the automation of the test, but there are not recommendations from the international or national authorities of animal health regarding the procedure of the test in microplates. Furthermore, there are little reports on the performance and experimental conditions for the FPA test in microplate readers  ADDIN EN.CITE  ADDIN EN.CITE.DATA [9-11].
The aim of the present study was to adapt a commercially available kit for the FPA assay to the 96-well microplate format, using sera from free herds and endemic areas of Argentina. Different dilutions of both serum and commercial antigen were assayed, and the polarization of the fluorescence was obtained with a multimode microplate reader. Once the optimal conditions for the FPA assay were established, its diagnostic performance was compared to that of the conventional agglutination tests in sera from an endemic area of Argentina.

Material and methods

2.1. Samples
Serum samples were obtained from 20 bulls and 36 cows belonging to free herds where vaccination of female calves between 3 and 8 month of age is the routine. One hundred and thirteen serum samples were obtained from herds located in an endemic area (province of Santiago del Estero, Argentina). Samples were stored at -20�C until used.

2.2. Fluorescence polarization assay
The antigen, the diluent of sera and the negative and positive control sera were provided in the FPA kit produced by Laboratorio Biol�gico Tandil (Tandil, Buenos Aires, Argentina). The antigen was composed by the O-polysaccharide from B. abortus 1119-3 conjugated with fluorescein isothiocyanate. All the components of the kit were produced according to the recommendations published by the OIE, on the basis of the protocol of Dr Klaus Nielsen. 
The test was carried out in 96-well black polypropylene microplates (Greiner Bio One, Frickenhausen, Germany) in a final volume of 200 �l. Briefly, serum samples were clarified by centrifugation (2 min at 10000 rpm). Appropriate dilution of sera was prepared directly in the wells of the microplate and a blank reading was performed. When antigen was used at a 1:100 final dilution, a 1:10 pre-dilution was prepared as to avoid pippeting low volumes of antigen; when final dilution of antigen was 1:20, it was used directly from the undiluted stock. The test mixture was incubated with orbital shaking for 2 min, and after two additional min of incubation the polarization of the fluorescence was measured. A Beckman multimode reader (Model DTX 880, Beckman Coulter, Brea, California) with Multimode Detection Software version 2.1.0.17 was used. DTX parameters used for FPA measurements were 485nm excitation, 535nm emission and 0.65 gain. Results of total polarization obtained with blanks (diluted serum) and the total reaction mixture (diluted serum plus antigen) were exported to an Excel� spreadsheet (Microsoft Corp, Redmond, Washington) in which the following formula was used to calculate the mP value:
mP= (Vt-Vb) � G(Ht-Hb)
         (Vt-Vb) + G(Ht-Hb)
where Vt = vertical intensity of the test after the tracer was added; Vb = vertical intensity of the test before the tracer was added; Ht = horizontal intensity of the test after the tracer was added; Hb = horizontal intensity of the test before the tracer was added; G = G factor =0.65  ADDIN EN.CITE  ADDIN EN.CITE.DATA [9]. The criteria for interpretation of results were according to that established by the national authorities of animal health, SENASA (Servicio Nacional de Sanidad Animal y Calidad Agroalimentaria): positive if mP was e"105 mP, negative if the value was <94 mP and indeterminate when the mP value was between 94 and 105.

2.3. Agglutination tests
Conventional agglutination tests (BPAT and SAT/b�-ME) were carried out and interpreted according to the procedures recommended by the SENASA  ADDIN EN.CITE <EndNote><Cite><Author>Nicola</Author><Year>2009</Year><RecNum>48</RecNum><record><rec-number>48</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">48</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Nicola, A. M.</author><author>Elena, S.</author></authors></contributors><titles><title>Manual de diagn�stico serol�gico de la brucelosis bovina&#xD;</title><secondary-title>Servicio Nacional de Sanidad Animal. Argentina</secondary-title></titles><periodical><full-title>Servicio Nacional de Sanidad Animal. Argentina</full-title></periodical><dates><year>2009</year></dates><urls></urls></record></Cite></EndNote>[12]. Antigens were obtained from Laboratorio Biol�gico Tandil (Tandil, Argentina). BPAT was used as screening test, and positive samples were further tested in SAT with and without b�-ME in parallel. Table 1 shows the criteria for the interpretation of results of these tests in cows older than 18 months that were vaccinated as calves with B abortus S19. Antigen and control sera for these tests were provided by Laboratorio Biol�gico de Tandil.

2.4. Statistical Analyses
The comparisons of test conditions for the FPA was analyzed by McNemar�s t test for paired data, using the GraphPad online calculator, 2002-2005 by GraphPad Software Inc. available at HYPERLINK "http://www.graphpad.com/quickcalcs/index.cfm"http://www.graphpad.com/quickcalcs/index.cfm. 

Results

In order to set the optimal conditions to perform the FPA assay in microplates, we first test some experimental conditions and settings of the equipment including the volume of total reaction and the integration time of the polarimeter. We did not found statistically significant differences in the results when performing the reaction in 100 or 200�l of final volume, nor in the setting of the integration time between 0.05 and 1.25 sec (data not shown). We then set the final volume in 200�l and the integration time in 0.25 sec, and compared different proportions of both serum and antigen. Preliminary experiments with a limited number of samples showed no significant differences in the mP units obtained when antigen and sera were combined in different proportions (data not shown). To decide which proportion of serum and antigen performed better, we then compare, using a panel of 144 samples, the conditions recommended by the SENASA and OIE for the test performed in 10mm x 75mm borosilicate glass tubes (serum and antigen at a 1:100 dilution) with that used by Minas et al  ADDIN EN.CITE  ADDIN EN.CITE.DATA [10] in sheep and goat sera and McGiven et al  ADDIN EN.CITE  ADDIN EN.CITE.DATA [11] in bovine sera (serum at a 1:10 dilution and antigen at 1:20).
Six from the 144 serum samples tested in both conditions gave indeterminate result in any of the conditions tested and were excluded from the comparison. Table 2 shows the results obtained from the comparison of samples tested using the two different proportions of reactants. Ninety seven percent (135/138) gave concordant result in the two variants of the assay, while 3 samples resulted positive only when tested at 1:100 dilution. These differences were not statistically significant (Mc Nemar�s test, p=0.2482). The 3 samples that resulted positive at 1:100 dilution were also positive in the conventional agglutination tests, applying the criteria for interpretation of the results currently used in Argentina (title e" 200 in SAT or e" 50 incomplete in b�-�ME, Table 1).
We then set the dilution at 1:100 for both serum and antigen for the comparison of the performance of the FPA assay with the conventional agglutination tests. Results from this comparison are summarized in Table 3. Concordance between results from the agglutination tests and FPA was observed in 94 from 113 samples (83.2%). Seventeen from the 18 samples giving discordant results (positive or suspected by the agglutination tests and negative in the FPA) were also tested by FPA using sera at a 1:10 dilution and antigen at 1:20 and resulted negative (the remainder sample could not be tested in this latter condition because of insufficient volume). The percentage of false positive reactions observed in the conventional agglutination tests compared to FPA was 8.8%. Eigth percent of samples giving suspected result in the agglutination tests, which should be re-tested in order to give a definitive result, were defined as negative in the FPA.


Discussion

Although the FPA test has been recommended by the SENASA, most accredited laboratories still use the conventional agglutination tests for screening and confirmation of seroreactivity to B. abortus, mainly because of the lack of the necessary equipment. Nowadays, the acquisition of multimode microplate readers including the fluorescence polarization mode of reading is being considered, as this equipment can be employed for many applications. Most fluorescence polarization analyzers have fixed reading parameters, while they can be modified in the microplate analyzers, like the Beckman Coulter DTX-880 we have used in the present study.

We compared two different proportions of the two reagents, serum and antigen, because we have not found official recommendations (OIE or SENASA in Argentina) for the procedure of the test in microplates. As to that we have found in the literature, there is only one study in which the FPA was conducted in microplates with cattle sera  ADDIN EN.CITE  ADDIN EN.CITE.DATA [11]. In two other studies the FPA was carried out in microplates using sheep and goat sera  ADDIN EN.CITE  ADDIN EN.CITE.DATA [10,13]. In one of these studies the authors used the 1:100 dilution of both reagents while in the others serum was used at 1:10 and antigen at 1:20. Our results show that the proportions currently used for the procedure in glass tubes (dilution 1:100 for both the serum and antigen) performed as well as the variant with antigen at 1:20 and serum at 1:10. The main advantage of using the former option is that there is an economy of 80% in antigen.

While the conventional agglutination tests have widely contributed to the control of the disease, they have some disadvantages like subjectivity, they are time-consuming, and may give false positive results due to cross reaction with other Gram negative bacteria and residual antibodies generated by vaccination  ADDIN EN.CITE  ADDIN EN.CITE.DATA [5-7].
The FPA has been first validated using samples from Canada where brucellosis has been eradicated and vaccination with B.abortus S19 is rare. The cut off has been adapted for the testing of samples from Argentina, where cattle is vaccinated between 3 and 8 month of age  ADDIN EN.CITE <EndNote><Cite><Author>Nielsen</Author><Year>1998</Year><RecNum>33</RecNum><record><rec-number>33</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">33</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Nielsen, K.</author><author>Gall, D.</author><author>Lin, M.</author><author>Massangill, C.</author><author>Samartino, L.</author><author>Perez, B.</author><author>Coats, M.</author><author>Hennager, S.</author><author>Dajer, A.</author><author>Nicoletti, P.</author><author>Thomas, F.</author></authors></contributors><auth-address>Canadian Food Inspection Agency, Animal Diseases Research Institute, Nepean, Ont., Canada. nielsenk@em.agr.ca</auth-address><titles><title>Diagnosis of bovine brucellosis using a homogeneous fluorescence polarization assay</title><secondary-title>Vet Immunol Immunopathol</secondary-title></titles><periodical><full-title>Vet Immunol Immunopathol</full-title></periodical><pages>321-9</pages><volume>66</volume><number>3-4</number><edition>1999/01/08</edition><keywords><keyword>Animals</keyword><keyword>Brucella abortus/immunology</keyword><keyword>Brucellosis, Bovine/*diagnosis/prevention &amp; control</keyword><keyword>Cattle</keyword><keyword>Fluorescence Polarization/methods/*veterinary</keyword><keyword>Sensitivity and Specificity</keyword><keyword>Software</keyword></keywords><dates><year>1998</year><pub-dates><date>Dec 11</date></pub-dates></dates><isbn>0165-2427 (Print)&#xD;0165-2427 (Linking)</isbn><accession-num>9880108</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=9880108</url></related-urls></urls><electronic-resource-num>S0165-2427(98)00195-0 [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>[7]. Samartino et al [14] evaluated the performance of the FPA in samples from argentinian cattle in which the serological status was defined according to its reaction in BPAT and competitive ELISA, finding that the FPA aided in the definition of samples that had discordant results between BPAT and ELISA. 
The fact that FPA was negative in all samples giving suspected result in the conventional agglutination tests is beneficial as it avoids the need to re-test these samples that, in our study, accounted for an 8% of the total. An additional 8% of samples which were positive in the agglutination tests and resulted negative in the FPA are considered false positive reactors. False positive reaction in agglutination test are well documented  ADDIN EN.CITE <EndNote><Cite><Author>Emmerzaal</Author><Year>2002</Year><RecNum>54</RecNum><record><rec-number>54</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">54</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Emmerzaal, A.</author><author>de Wit, J. J.</author><author>Dijkstra, T.</author><author>Bakker, D.</author><author>van Zijderveld, F. G.</author></authors></contributors><auth-address>Animal Health Service, Deventer, The Netherlands.</auth-address><titles><title>The Dutch Brucella abortus monitoring programme for cattle: the impact of false-positive serological reactions and comparison of serological tests</title><secondary-title>Vet Q</secondary-title></titles><periodical><full-title>Vet Q</full-title></periodical><pages>40-6</pages><volume>24</volume><number>1</number><edition>2002/04/02</edition><keywords><keyword>Agglutination Tests</keyword><keyword>Animal Husbandry</keyword><keyword>Animals</keyword><keyword>Antibodies, Bacterial/analysis</keyword><keyword>Brucella abortus/*immunology</keyword><keyword>Brucellosis, Bovine/*diagnosis/immunology</keyword><keyword>Cattle</keyword><keyword>Complement Fixation Tests</keyword><keyword>Enzyme-Linked Immunosorbent Assay</keyword><keyword>False Positive Reactions</keyword><keyword>Netherlands</keyword><keyword>Sensitivity and Specificity</keyword><keyword>Serologic Tests</keyword></keywords><dates><year>2002</year><pub-dates><date>Feb</date></pub-dates></dates><isbn>0165-2176 (Print)&#xD;0165-2176 (Linking)</isbn><accession-num>11924560</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=11924560</url></related-urls></urls><electronic-resource-num>10.1080/01652176.2002.9695123</electronic-resource-num><language>eng</language></record></Cite></EndNote>[14] and mainly due to cross reaction with antibodies to other Gram negative bacteria like Yersinia enterocol�tica, Pasteurella multocida and Escherichia coli  ADDIN EN.CITE  ADDIN EN.CITE.DATA [15,16]. In a study with serum samples from Argentina, Samartino et al  ADDIN EN.CITE <EndNote><Cite><Author>Samartino</Author><Year>1999</Year><RecNum>31</RecNum><record><rec-number>31</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">31</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Samartino, L.</author><author>Gregoret, R.</author><author>Gall, D.</author><author>Nielsen, K.</author></authors></contributors><auth-address>CICV-INTA, Instituto de Patholobiologia, Buenos Aires, Argentina.</auth-address><titles><title>Fluorescence polarization assay: application to the diagnosis of bovine brucellosis in Argentina</title><secondary-title>J Immunoassay</secondary-title></titles><periodical><full-title>J Immunoassay</full-title></periodical><pages>115-26</pages><volume>20</volume><number>3</number><edition>1999/08/24</edition><keywords><keyword>Animals</keyword><keyword>Antibodies, Bacterial/blood</keyword><keyword>Argentina/epidemiology</keyword><keyword>Brucellosis, Bovine/*diagnosis/epidemiology/prevention &amp; control</keyword><keyword>Cattle</keyword><keyword>Fluorescence Polarization Immunoassay</keyword><keyword>Sensitivity and Specificity</keyword><keyword>Vaccination/veterinary</keyword></keywords><dates><year>1999</year><pub-dates><date>Aug</date></pub-dates></dates><isbn>0197-1522 (Print)&#xD;0197-1522 (Linking)</isbn><accession-num>10458638</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=10458638</url></related-urls></urls><electronic-resource-num>10.1080/01971529909349347</electronic-resource-num><language>eng</language></record></Cite></EndNote>[17] showed that specificity of the agglutination with the use of b�-mercaptoethanol was above 98% in cattle after nine month of vaccination with B. abortus S19. Then, as the b�-ME test was used in the sequence of tests carried out in the present study, it seems likely that the false positive reactions were mainly due to cross reaction with other Gram negative bacteria.
FPA performed in microplates increases testing capacity, is an accurate diagnostic test that has many advantages compared with other methods used for serological diagnosis of bovine brucellosis; and our results show that it is also possible to use less antigen than that used in the scarce previous reports.

Acknowledgments
This work was supported by grants from the Agencia Nacional de Promoci�n Cient�fica y Tecnol�gica (PAE-PICT 2007-24 to SEG, ANPCYT-Argentina). AGD is a fellow from CONICET (Argentina). SEG and SME are members of the Research Career of CONICET. All authors read and approved the final manuscript.
References

 ADDIN EN.REFLIST 1.	Aznar, MN, Samartino, LE, Humblet, MF Saegerman, C (2012) Bovine Brucellosis in Argentina and Bordering Countries: Update. Transbound Emerg Dis. doi: 10.1111/tbed.12018.
2.	Schurig, GG, Roop, RM, 2nd, Bagchi, T, Boyle, S, Buhrman, D et al. (1991) Biological properties of RB51; a stable rough strain of Brucella abortus. Vet Microbiol 28: 171-188.
3.	Nielsen, K, Smith, P, Gall, D, Perez, B, Cosma, C et al. (1996) Development and validation of an indirect enzyme immunoassay for detection of antibody to Brucella abortus in milk. Vet Microbiol 52: 165-173.
4.	Nielsen, K, Smith, P, Widdison, J, Gall, D, Kelly, L et al. (2004) Serological relationship between cattle exposed to Brucella abortus, Yersinia enterocolitica O:9 and Escherichia coli O157:H7. Vet Microbiol 100: 25-30.
5.	Nielsen, K, Gall, D, Jolley, M, Leishman, G, Balsevicius, S et al. (1996) A homogeneous fluorescence polarization assay for detection of antibody to Brucella abortus. J Immunol Methods 195: 161-168.
6.	Aguirre, NP, Vanzini, VR, Torioni de Echaide, S, Valentini, BS, De Lucca, G et al. (2002) Antibody dynamics in Holstein Friesian heifers vaccinated with Brucella abortus strain 19, using seven serological tests. J Immunoassay Immunochem 23: 471-478.
7.	Nielsen, K, Gall, D, Lin, M, Massangill, C, Samartino, L et al. (1998) Diagnosis of bovine brucellosis using a homogeneous fluorescence polarization assay. Vet Immunol Immunopathol 66: 321-329.
8.	OIE. (2009) Chapter 2.4.3.:Bovine brucellosis. In Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. , Office International des Epizooties, Paris.
9.	Ramirez-Pfeiffer, C, Diaz-Aparicio, E, Rodriguez-Padilla, C, Morales-Loredo, A, Alvarez-Ojeda, G et al. (2008) Improved performance of Brucella melitensis native hapten over Brucella abortus OPS tracer on goat antibody detection by the fluorescence polarization assay. Vet Immunol Immunopathol 123: 223-229.
10.	Minas, A, Stournara, A, Minas, M, Stack, J, Petridou, E et al. (2007) Validation of a fluorescence polarization assay (FPA) performed in microplates and comparison with other tests used for diagnosing B. melitensis infection in sheep and goats. J Immunol Methods 320: 94-103.
11.	McGiven, JA, Tucker, JD, Perrett, LL, Stack, JA, Brew, SD et al. (2003) Validation of FPA and cELISA for the detection of antibodies to Brucella abortus in cattle sera and comparison to SAT, CFT, and iELISA. J Immunol Methods 278: 171-178.
12.	Nicola, AMElena, S (2009) Manual de diagn�stico serol�gico de la brucelosis bovina
Servicio Nacional de Sanidad Animal. Argentina.
13.	Ramirez-Pfeiffer, C, Diaz-Aparicio, E, Gomez-Flores, R, Rodriguez-Padilla, C, Morales-Loredo, A et al. (2008) Use of the Brucella melitensis native hapten to diagnose brucellosis in goats by a rapid, simple, and specific fluorescence polarization assay. Clin Vaccine Immunol 15: 911-915.
&'AQ������������°���zh��YI9h�hMLCJH*aJmH	sH	h�h8QCJH*aJmH	sH	h�h8QCJaJmH	sH	"h�h,�5�CJ\�aJmH	sH	hC^�5�CJ\�aJmH	sH	"h�h�1�5�CJ\�aJmH	sH	(h�ht�5�6�CJ\�]�aJmH	sH	"h�ht�5�CJ\�aJmH	sH	h�h�1�5�CJaJmH	sH	h�ht�5�CJaJmH	sH	h�h�m�CJaJmH	sH	h�h�1�CJaJmH	sH	���		�
�
�
�
Rq������MN\���������������������������d�gd62�d�gd�5>d�gd�<7d�gdMLd�gd�N�����	
								 	;	<	�	�	�	�	�	v
�
�
�
�
�
�
�
�
�
���������ѥ����������|��peZeh�h�1�CJaJh�hMLCJaJh�hMLCJH*aJh2}CJaJh�hOK"CJaJh�h�O]CJaJh�h8QCJaJh�h8QCJH*aJh�hMLCJH*aJmH	sH	h�h8QCJH*aJmH	sH	h�h8QCJaJmH	sH	h�hMLCJaJmH	sH	h�h�<7CJH*aJmH	sH	�
�
�
�
�
�
�
&Rq�����������������Ժ����r�cSG8G8Gh�h�5>CJaJmH	sH	h�e�CJaJmH	sH	h�h�5>5�CJaJmH	sH	h�hMLCJaJmH	sH	$h�ZYh�<7CJPJ^JaJmH	sH	,h�ZYh�<7CJOJPJQJ^JaJmH	sH	h�ZYh2}CJaJmH	sH	h�ZYh�05CJaJmH	sH	h�ZYh�<7CJaJmH	sH	h�h�05CJaJh�h�<7CJaJh�ZYh��CJaJh�ZYhOK"CJaJh�ZYh�<7CJaJ���%0156abghpqr~�������X
m
yz�������ʻʻʻʪʞ��ʻʞʞʐ��s�c�Th�h�BCJaJmH	sH	h�h�5>6�CJaJmH	sH	hz�CJaJmH	sH	 h�h�5>CJ^JaJmH	sH	h�R%CJ^JaJmH	sH	h�p�CJaJmH	sH	 h�h�R%CJ^JaJmH	sH	h�h�R%CJaJmH	sH	h�R%CJaJmH	sH	h�h�5>CJaJmH	sH	h�e�CJaJmH	sH	h�h�e�CJaJmH	sH	 �� 8�����N\_abcnop{|������ϰ���qbSG�bGb�9�h�RCJ^JaJmH	sH	h�RCJaJmH	sH	h�h�n�CJaJmH	sH	h�h�BCJaJmH	sH	h�hL�5�CJaJmH	sH	h�h62�CJaJmH	sH	h�h�5�CJaJmH	sH	h�h62�5�CJaJmH	sH	 h�hL�CJ^JaJmH	sH	h�x�CJ^JaJmH	sH	 h�h�5>CJ^JaJmH	sH	h�h�5>CJaJmH	sH	 h�h�BCJ^JaJmH	sH	���������������������,0@BPR\^�����ŷŷ�������ŷ�����sesesWh��CJ^JaJmH	sH	hz�CJ^JaJmH	sH	 h�h��CJ^JaJmH	sH	$h�h�BCJOJQJaJmH	sH	 h�h�n�CJ^JaJmH	sH	h�h�n�CJaJmH	sH	h�RCJ^JaJmH	sH	 h�h�BCJ^JaJmH	sH	h�RCJaJmH	sH	h�h�BCJaJmH	sH	h�hL�CJaJmH	sH	 ��0^`|~G$X3r5s5�5�5�5�6�67�8������������������d�gd�C��l�dh]�l�gdqMd�gd�Md�gd�h�d�gd�*�d�7$8$H$gd�*�d�7$8$H$gd�J�d�gd�J�
&Fd�gdTyd�gd�5>^`bz|~�������ò�tU�6<h�h�97B*CJKH$PJ\�^JaJmH	nH
,phsH	tH
,<h�h1iB*CJKH$PJ\�^JaJmH	nH
,phsH	tH
,<h�h}B*CJKH$PJ\�^JaJmH	nH
,phsH	tH
,<h�h1�B*CJKH$PJ\�^JaJmH	nH
,phsH	tH
, h�h1o�CJ^JaJmH	sH	h�h�1�5�CJaJmH	sH	h�h�m�5�CJaJmH	sH	h�h�5>CJaJmH	sH	h�o�CJ^JaJmH	sH	
�����12bcfg�£ƒcD�D�D�(�D6h>.B*CJKH$PJ\�^JaJmH	nH
,phsH	tH
,<h�h�rVB*CJKH$PJ\�^JaJmH	nH
,phsH	tH
,?h�h1�6�B*CJKH$PJ\�^JaJmH	nH
,phsH	tH
,?h�h�976�B*CJKH$PJ\�^JaJmH	nH
,phsH	tH
,<h�h�GPB*CJKH$PJ\�^JaJmH	nH
,phsH	tH
,<h�h�97B*CJKH$PJ\�^JaJmH	nH
,phsH	tH
,<h�h��B*CJKH$PJ\�^JaJmH	nH
,phsH	tH
,gqr���EFRu��������{\<\?h�h
�6�B*CJKH$PJ\�^JaJmH	nH
,phsH	tH
,<h�h
�B*CJKH$PJ\�^JaJmH	nH
,phsH	tH
, h�h
�CJ^JaJmH	sH	 h�h�`7CJ^JaJmH	sH	 h�hlJkCJ^JaJmH	sH	 h�h}CJ^JaJmH	sH	 h�h�rVCJ^JaJmH	sH	 h�h,�CJ^JaJmH	sH	<h�h�GPB*CJKH$PJ\�^JaJmH	nH
,phsH	tH
,���������������£�g�g��H7&7 h�htHCJ^JaJmH	sH	 h�h,�CJ^JaJmH	sH	<h�h�&�B*CJKH$PJ\�^JaJmH	nH
,phsH	tH
,6h�$�B*CJKH$PJ\�^JaJmH	nH
,phsH	tH
,?jh�6
B*CJKH$PJU\�^JaJmH	nH
,phsH	tH
,<h�h�z�B*CJKH$PJ\�^JaJmH	nH
,phsH	tH
,<h�h�}�B*CJKH$PJ\�^JaJmH	nH
,phsH	tH
,<h�hwYgB*CJKH$PJ\�^JaJmH	nH
,phsH	tH
,
����FQTu������� ���޿ޮޚ�މ�{j{Y{�K�K�Kh�,sCJ^JaJmH	sH	 h�h�
ECJ^JaJmH	sH	 h�h��CJ^JaJmH	sH	h�
ECJ^JaJmH	sH	 h�htHCJ^JaJmH	sH	&h�h,�6�CJ]�^JaJmH	sH	 h�h�CJ^JaJmH	sH	h�p�CJ^JaJmH	sH	 h�h�`7CJ^JaJmH	sH	 h�h,�CJ^JaJmH	sH	 h�h,�CJ^JaJmH	sH	 *QTVWbgk  & . 0 6 8 � � � � � � � � � 8!<!������ᮙ�zez��TC����C� h�hmM*CJ^JaJmH	sH	 h�htHCJ^JaJmH	sH	(h�h)A�CJOJQJ^JaJmH	sH	 h�h)A�CJ^JaJmH	sH	h)A�CJ^JaJmH	sH	(h�hwYgCJOJQJ^JaJmH	sH	 h�hwYgCJ^JaJmH	sH	 h�h�<#CJ^JaJmH	sH	 h�h�n�CJ^JaJmH	sH	 h�h,�CJ^JaJmH	sH	h�^�CJ^JaJmH	sH	<!D!P!^!�!""""*"+"?"@"A"B"G"H"I"J"�"�"
#<#F#����ͻͩ���t�����_�N�N<#h�hT�6�CJ^JaJmH	sH	 h�hT�CJ^JaJmH	sH	)h�h,�B*CJ^JaJmH	phsH	)�jh�$�CJU^JaJmH	sH	#jh�$�CJU^JaJmH	sH	h�$�CJ^JaJmH	sH	#jh�oCJU^JaJmH	sH	#h�h�6�CJ^JaJmH	sH	 h�h�CJ^JaJmH	sH	 h�h,�CJ^JaJmH	sH	 h�hmM*CJ^JaJmH	sH	F#�#�#�#�#�#�#�#�#�#$$$$$$$$k$u$v$�$�����о���������wfTfC h�h�BCJ^JaJmH	sH	#h�h,�6�CJ^JaJmH	sH	 h�h,�CJ^JaJmH	sH	#h�h,�7�CJ^JaJmH	sH	)�j�h�$�CJU^JaJmH	sH	#jh�$�CJU^JaJmH	sH	h�$�CJ^JaJmH	sH	#jh�[�CJU^JaJmH	sH	hf.CJ^JaJmH	sH	 h�htHCJ^JaJmH	sH	 h�hT�CJ^JaJmH	sH	�$�$�$e%j%k%l%�+�+�+�+�+�++,5,j,n,�,�,�,�,�,�,�,��оЬ����ооЍ�|kZK<�h�h�J&CJaJmH	sH	h�hP\CJaJmH	sH	 h�hP\CJ^JaJmH	sH	 h�h�J&CJ^JaJmH	sH	 h�hT�CJ^JaJmH	sH	 h�h4m�CJ^JaJmH	sH	h�$�CJ^JaJmH	sH	#jhf.CJU^JaJmH	sH	#h�h,�6�CJ^JaJmH	sH	 h�h,�CJ^JaJmH	sH	 h�h�BCJ^JaJmH	sH	hz�CJ^JaJmH	sH	�,�,�,�,�,�,�,�,�,---	-
---.-�����౟�}�fWCW&jh�$�6�CJU^JaJmH	sH	h�$�6�CJ^JaJmH	sH	,jh�:h�:6�CJU^JaJmH	sH	h�hP\6�CJaJmH	sH	#h�hP\6�CJ^JaJmH	sH	#h�h,�6�CJ^JaJmH	sH	 h�h,�CJ^JaJmH	sH	h�h�J&CJaJmH	sH	h�hX2?CJaJmH	sH	h�hP\CJaJmH	sH	 h�hP\CJ^JaJmH	sH	.-/-0-1-6-7-8-9-o-�-�-[.a.j.�վ����{gVA,A)h�h{�B*CJ^JaJmH	phsH	)h�h,�B*CJ^JaJmH	phsH	 h�h,�CJ^JaJmH	sH	'h�h�E�CJPJaJmHnHsH	u h�h�E�CJ^JaJmH	sH	 h�k�h,�CJ^JaJmH	sH	#h�:h,�6�CJ^JaJmH	sH	h�$�6�CJ^JaJmH	sH	,jh�:h�:6�CJU^JaJmH	sH	&jh�$�6�CJU^JaJmH	sH	,�j�h�$�6�CJU^JaJmH	sH	
j.k.l.�0�0�0�0,141;1h1�1�1�1�1�1�1�ооЩ��p�[�F1�)h�hqMB*CJ^JaJmH	phsH	)h�h�*�B*CJ^JaJmH	phsH	)h�h"Z�B*CJ^JaJmH	phsH	)h�hlJkB*CJ^JaJmH	phsH	h�h�E�CJaJmH	sH	)h�h�E�B*CJ^JaJmH	phsH	)h�h,�B*CJ^JaJmH	phsH	#h�$�B*CJ^JaJmH	phsH	2jh�h%@�B*CJU^JaJmH	phsH	)h�h%@�B*CJ^JaJmH	phsH	�1�1�12L2i2j2o2q2�2�2�2�2&3'3(3��嵤������x�]>=jh�h�oB*CJPJU^JaJmHnHphsH	u4h�h�MB*CJPJ^JaJmHnHphsH	u4h�h4m�B*CJPJ^JaJmHnHphsH	u h�h�MCJ^JaJmH	sH	 h�hz�CJ^JaJmH	sH	4h�hz�B*CJPJ^JaJmHnHphsH	u)h�h,�B*CJ^JaJmH	phsH	4h�h,�B*CJPJ^JaJmHnHphsH	u(37383L3M3N3O3U3V3W3|3�3�3�3�3�3�3�3S4T4_4���̎�s^I^4^I^4^4^)h�h"Z�B*CJ^JaJmH	phsH	)h�h4m�B*CJ^JaJmH	phsH	)h�h,�B*CJ^JaJmH	phsH	4h�h,�B*CJPJ^JaJmHnHphsH	u=jh�h�oB*CJPJU^JaJmHnHphsH	u=�j�'h�$�B*CJPJU^JaJmHnHphsH	u7jh�$�B*CJPJU^JaJmHnHphsH	u.h�$�B*CJPJ^JaJmHnHphsH	u_4�4r5s5�5�5�5�5�5�5�5�5�5�536�6�6�п���n]L;L]*n h�h�h4CJ^JaJmH	sH	 h�h�MCJ^JaJmH	sH	 h�hS2WCJ^JaJmH	sH	 h�h�CJ^JaJmH	sH	 h�h3CJ^JaJmH	sH	 h�hTyCJ^JaJmH	sH	h�h�6CJaJmH	sH	h_!h35�CJaJmH	sH	h_!h�<#5�CJaJmH	sH	 h�h,�CJ^JaJmH	sH	)h�h,�B*CJ^JaJmH	phsH	4h�h,�B*CJPJ^JaJmHnHphsH	u�6�6�6�677,7-757�7�7�7�7�7�7888)8*838�8��ͼ�͚ͫ��z��f�U�D�� h�h�g!CJ^JaJmH	sH	 h�h2zqCJ^JaJmH	sH	&h�h8-6�CJ]�^JaJmH	sH	h�h�MCJaJmH	sH	 h�h�MCJ^JaJmH	sH	 h�h8-CJ^JaJmH	sH	 h�hS2WCJ^JaJmH	sH	 h�h�<#CJ^JaJmH	sH	 h�h�%:CJ^JaJmH	sH	 h�hTyCJ^JaJmH	sH	 h�hd�CJ^JaJmH	sH	�8�8�8�8�8�8�8�899!97999Q9S9_9�9�9
::::���ͼͼ��͌�{j{YHYH7Y h�h�HCJ^JaJmH	sH	 h�h�R�CJ^JaJmH	sH	 h�h�CJ^JaJmH	sH	 h�h�<#CJ^JaJmH	sH	 h�h�4kCJ^JaJmH	sH	huk�CJ^JaJmH	sH	 h�hd�CJ^JaJmH	sH	 h�h2zqCJ^JaJmH	sH	 h�h��CJ^JaJmH	sH	 h�h1|cCJ^JaJmH	sH	 h�h�h4CJ^JaJmH	sH	 h�h�1CJ^JaJmH	sH	:::@:b:j:z:{:�:�:�:�:�:�:�:�:�:�:�:.;0;\;�;�;�;�;�;�;�;�;�;�;�;�;�;�;������������������~p~_~� h�h�b�CJ^JaJmH	sH	hD-�CJ^JaJmH	sH	h�b�CJ^JaJmH	sH	 hD-�h�i�CJ^JaJmH	sH	 h�h�i�CJ^JaJmH	sH	 h�h2zqCJ^JaJmH	sH	 h�hS2WCJ^JaJmH	sH	ht �CJ^JaJmH	sH	 h�h�R�CJ^JaJmH	sH	 h�h�HCJ^JaJmH	sH	"�;<<<8<Y<m<n<w<===='=A=�=�=�=�=�=���ͼ͚ͫ�x��j�Y�G3&h�h�$:>*CJH*^JaJmH	sH	#h�h�$:>*CJ^JaJmH	sH	 h�h�1CJ^JaJmH	sH	ht �CJ^JaJmH	sH	 h�h�R�CJ^JaJmH	sH	 h�h�$:CJ^JaJmH	sH	 h�h�CJ^JaJmH	sH	 h�h2zqCJ^JaJmH	sH	 h�h1|cCJ^JaJmH	sH	 h�h�CJ^JaJmH	sH	 h�h�HCJ^JaJmH	sH	 h�h�YCJ^JaJmH	sH	�8�=�=�=�@�@A	H
H$HFIGIOIPIPNfR�V�V�V������������������d�7$8$H$gd�d�gd�y�d�7$8$H$gdyz�
&Fd�7$8$H$gdyz�d�gd�h�d�7$8$H$gd�h�d�gd�h��=�=�=�=�=�=�=�=�=�=�=�=�=�=�=�=�=�=�=�=�=>>A>B>e>f>p>w>�>�>�>�>�>�>�>�������ȶȶȶȶȥ����������������m_h�$�CJ^JaJmH	sH	)jh�h�CJU^JaJmH	sH	 h�h2zqCJ^JaJmH	sH	#h�h��CJH*^JaJmH	sH	 h�h��CJ^JaJmH	sH	#h�h�$:CJH*^JaJmH	sH	 h�h�$:CJ^JaJmH	sH	&h�h�$:>*CJH*^JaJmH	sH	#h�h�$:>*CJ^JaJmH	sH	#�>�>�>�>�>???1?6?W?b?�?�?�?�?�?@@F@H@�@�@����ߵ����sb���bQbQb@ h�h�$:CJ^JaJmH	sH	 h�h�S�CJ^JaJmH	sH	 h�h�?�CJ^JaJmH	sH	h�h�RwCJaJmH	sH	 h�h�RwCJ^JaJmH	sH	 h�h9>�CJ^JaJmH	sH	 h�h�4kCJ^JaJmH	sH	)jh�h�CJU^JaJmH	sH	)�jAh�$�CJU^JaJmH	sH	h�$�CJ^JaJmH	sH	#jh�$�CJU^JaJmH	sH	�@�@�@�@�@�@�@A>A@AJALA^A`ABB
B�D�D�D�D�D��ͼͫޫ������xcUcUcD h�h�Z~OJQJ^JmH	sH	h�$�OJQJ^JmH	sH	)jh�h�OJQJU^JmH	sH	 h�h�*�OJQJ^JmH	sH	 h�hfnOJQJ^JmH	sH	 h�h�RwOJQJ^JmH	sH	 h�h�9�OJQJ^JmH	sH	 h�h9>�OJQJ^JmH	sH	 h�h�?�OJQJ^JmH	sH	 h�hTyOJQJ^JmH	sH	 h�h�9�CJ^JaJmH	sH	�D�D�D)E*E-E.EFF$F�F�F�F
G G&G>GPG�G�G�G�G�G���Ƶ�����q�`�`�N�`�=� h�h�b�OJQJ^JmH	sH	#h�h�Z~6�OJQJ^JmH	sH	 h�h�OJQJ^JmH	sH	 h�h�o*OJQJ^JmH	sH	 h�h�Z~OJQJ^JmH	sH	 h�h�F�OJQJ^JmH	sH	 h�h�RwOJQJ^JmH	sH	 h�h�
�OJQJ^JmH	sH	h�b�^JmH	sH	 hD-�h�b�OJQJ^JmH	sH	h�b�OJQJ^JmH	sH	 h.s�h�b�OJQJ^JmH	sH	�G�GHH	H
H$HZHdH�H�HIIBICIDIFIGIOI����ʻ���{�h�Y�J:h�h�?�5�CJaJmH	sH	h�h�?�CJaJmH	sH	h�h2#�CJaJmH	sH	$h�hxT�0JCJ^JaJmH	sH	h�h�CJaJmH	sH	jh�h�CJUaJ h�hxT�CJ^JaJmH	sH	 h�h�F�CJ^JaJmH	sH	h�h�F�CJaJmH	sH	h�h�RCJaJmH	sH	h�h�Z~mH	sH	 h�h�RwOJQJ^JmH	sH	h�h�RwmH	sH	OIPIJJ=J>JNJXJhJtJ�J�J�J;K<KCKbKcKpKqK�K�K�K�K�KuL�L�L�����ⵤ�Ĥ�Ӥӕ��ӕ��whwYH h�h�g~CJ^JaJmH	sH	h�h�g~CJaJmH	sH	h�hsG�CJaJmH	sH	h�h�CJaJmH	sH	h�h�jTCJaJmH	sH	h�h10nCJaJmH	sH	 h�h�;2CJ^JaJmH	sH	h�h�;2CJaJmH	sH	h�h�8�CJaJmH	sH	h�hk�CJaJmH	sH	h�h.CJaJmH	sH	h�h�LCJaJmH	sH	�L�L�L�L�L�L�L�L�L!M,M-M3M@MsM�M�M�M�M�M�M�M�M�M�M�M�M�M�MN��������޼޼�ޜ�ygyRg�y���ygy)�jrJh�$�CJU^JaJmH	sH	#jh�$�CJU^JaJmH	sH	h�$�CJ^JaJmH	sH	)jh�h�CJU^JaJmH	sH	 h�hwg�CJ^JaJmH	sH	h�h��CJaJmH	sH	 h�h��CJ^JaJmH	sH	 h�hx!CJ^JaJmH	sH	 h�h�g~CJ^JaJmH	sH	 h�h�jTCJ^JaJmH	sH	N	N
NNNN!NNNONPN�N�N�N�N
O9O=OeO��õä��sdUdFd7F7h�h�_�CJaJmH	sH	h�hsG�CJaJmH	sH	h�h�wYCJaJmH	sH	h�h�CJaJmH	sH	h�h�B�CJaJmH	sH	 h�h�}CJ^JaJmH	sH	 h�h�Q�CJ^JaJmH	sH	 h�hwg�CJ^JaJmH	sH	h�$�CJ^JaJmH	sH	)jh�h�CJU^JaJmH	sH	#jh�$�CJU^JaJmH	sH	)�j�Rh�$�CJU^JaJmH	sH	eO�O�O�O�O�O7PSPUPYP[PhPqP|P�PRR R"RFRJRfRvR~R�R�R�R�R�R��������ė��w�w�d�U�U�U�Uh�h/C�CJaJmH	sH	$h�h�CJOJQJaJmH	sH	 h�h�CJ^JaJmH	sH	h�h�CJaJmH	sH	h�h�CJaJmH	sH	h�h%hCJaJmH	sH	h�hKrsCJaJmH	sH	h�hsG�CJaJmH	sH	h�h�jTCJaJmH	sH	h�h�'BCJaJmH	sH	h�h^ �CJaJmH	sH	�RS�S�S�S�S�S�S�S/T�T�T�U�U�U�U�U�UVV2V3V��������ĵ�ӚӋ�whT@w'h�h0?�CJPJaJmHnHsH	u'h�h�`�CJPJaJmHnHsH	uh�h�`�CJaJmH	sH	'h�h�=�CJPJaJmHnHsH	uh�h}�CJaJmH	sH	h�h�=�CJaJmH	sH	h_!CJaJmH	sH	h�h�CJaJmH	sH	h�h�<CJaJmH	sH	h�h6H�CJaJmH	sH	h�h/C�CJaJmH	sH	h�hca�CJaJmH	sH	3VCVmV�V�V�V�V�V�V�V�V�V�V�V�V&W-WW�W�W�������륙�zj[L[=[h�hRT�CJaJmH	sH	h�h�7CJaJmH	sH	h�h>*CJaJmH	sH	h�hLnS5�CJaJmH	sH	h�h6H�5�CJaJmH	sH	h�h�5CJaJmH	sH	h�5CJaJmH	sH	h�h/C�CJaJmH	sH	'h�h,CJPJaJmHnHsH	uh�h�`�CJaJmH	sH	'h�h�=�CJPJaJmHnHsH	u'h�h%ytCJPJaJmHnHsH	u�V�V�VvYwY6]7]�^:h1|e}f}v}�~�~�~l��ρ����������������������$d�7$8$H$a$gd4�d�7$8$H$gd4�d�gd4�d�7$8$H$gdfZd�7$8$H$gd�h�
&Fd�7$8$H$gd�R�W�W�W�W�W�W�W�W�W�W�WXX0X5XwXxX�X�X�X�X�X�X�XuYvYwY����вУ����������v�g�gXIh�h�o�CJaJmH	sH	h�hB@CJaJmH	sH	h�h�:CJaJmH	sH	h�hkCJaJmH	sH	h�h.8CJaJmH	sH	h�hRT�CJaJmH	sH	h�h�[CJaJmH	sH	h�h�8�CJaJmH	sH	h�h��CJaJmH	sH	h�h>*CJaJmH	sH	h�hRT�6�CJaJmH	sH	h�h>*6�CJaJmH	sH	wY�Y�Y�YZCZoZ�Z�Z�Z�Z�Z�Z�Z�Z�Z�Z�Z�Z�Z
[1[6[J[�����ĵ����s����dU�U�FUh�hkCJaJmH	sH	h�hUI�CJaJmH	sH	h�hg�CJaJmH	sH	%�j>[h�$�CJUaJmH	sH	jh�$�CJUaJmH	sH	h�$�CJaJmH	sH	%jh�h�CJUaJmH	sH	h�h�L�CJaJmH	sH	h�h�:CJaJmH	sH	h�h�M{CJaJmH	sH	h�hRT�CJaJmH	sH	h�h�9NCJaJmH	sH	J[K[L[[[\[p[q[r[s[z[{[�[�[�[�[�[
\T\Z\5]6]7]@]A]����ү������񑂠�sdUFh�hRT�CJaJmH	sH	h�h��CJaJmH	sH	h�h7A
CJaJmH	sH	h�h�:CJaJmH	sH	h�hg�CJaJmH	sH	h�h�PCJaJmH	sH	h�hr>]CJaJmH	sH	%�j�ch�$�CJUaJmH	sH	jh�$�CJUaJmH	sH	h�$�CJaJmH	sH	%jh�h�CJUaJmH	sH	h�hWRCJaJmH	sH	A]L]a]�]
^B^K^c^o^p^^�^�^�^�^�^�^�^�^�^�^��οΰ�񝑁�n����_PAh�h�E�CJaJmH	sH	h�h��CJaJmH	sH	h�h�h�CJaJmH	sH	%�j>th�$�CJUaJmH	sH	jh�$�CJUaJmH	sH	h�$�CJaJmH	sH	%jh�h�CJUaJmH	sH	h�hr>]CJaJmH	sH	h�h$CJaJmH	sH	h�h��CJaJmH	sH	'h�hEO9CJPJaJmHnHsH	uh�hEO9CJaJmH	sH	�^�^�^�^__�_�_�_�_ggg	ggggg��κ���|cRcRc�C3Ch�h��6�CJaJmH	sH	h�h��CJaJmH	sH	!h�$�CJPJaJmHnHsH	u0jh�h�CJPJUaJmHnHsH	u'h�hHhCJPJaJmHnHsH	u'h�h��CJPJaJmHnHsH	u*h�h�m�6�CJPJaJmHnHsH	u'h�h�m�CJPJaJmHnHsH	u'h�h�E�CJPJaJmHnHsH	uh�h�E�CJaJmH	sH	h�h�m�CJaJmH	sH	gg g#g*gLghg�g�g�g�g�g�g�g�g�ghhhh2h3h�����®������u�aRaRAa h�hK�CJ^JaJmH	sH	h�h2�CJaJmH	sH	'h�h2�CJPJaJmHnHsH	u h�h�i�CJ^JaJmH	sH	'h�hU�CJPJaJmHnHsH	u'h�h
*CJPJaJmHnHsH	u'h�h�i�CJPJaJmHnHsH	u'h�h�LCJPJaJmHnHsH	uh�h�LCJaJmH	sH	h�h�m�CJaJmH	sH	hA0,CJaJmH	sH	3h4h6h9h:hHh�h�h�h�h�hi�i�i��dz���rcTcE6h�h"2_CJaJmH	sH	h�h��CJaJmH	sH	h�hLnSCJaJmH	sH	h�h
*CJaJmH	sH	h�hK�CJaJmH	sH	h�h��CJaJmH	sH	h�h)�CJaJmH	sH	'h�h)�CJPJaJmHnHsH	u'h�h�LCJPJaJmHnHsH	u'h�h2�CJPJaJmHnHsH	u'h�h�p�CJPJaJmHnHsH	u h�h2�CJ^JaJmH	sH	
�i�ioqpqtquq�q�q�q�q�q�q�q�q�q�q�q�q�q�qrrr r!r5r������³�Ѥ������u��ub�R�jh�$�CJUaJmH	sH	%jh�h�"!CJUaJmH	sH	h�hLMCJaJmH	sH	h�h�p�6�CJaJmH	sH	h�hLM6�CJaJmH	sH	h�hPyoCJaJmH	sH	h�h�p�CJaJmH	sH	h�htSCJaJmH	sH	h�h"2_CJaJmH	sH	h�$�CJaJmH	sH	%jh�h�CJUaJmH	sH	5r6r7r8r?r@rzrr�r�ry y$y%yzz�z�z�z�z�{�{0|1|V|��ɽɮ��������x���x�i�ZKh�hfZCJaJmH	sH	h�h�c�CJaJmH	sH	h�htSCJaJmH	sH	$h�hLMCJOJQJaJmH	sH	%jh�h�CJUaJmH	sH	h�hLM6�CJaJmH	sH	h�hLMCJaJmH	sH	h�$�CJaJmH	sH	%jh�h�"!CJUaJmH	sH	jh�$�CJUaJmH	sH	%�j�h�$�CJUaJmH	sH	V|W|�|�|
}}}}e}f}g}h}u}v}�}~	~~~~0~1~�~�~�~�~��������ĵ������y�jZKjh�h�CJUaJh�$�htS5�CJaJmH	sH	h�$�h~8�CJaJmH	sH	h�h�J"CJaJmH	sH	h�h�,PCJaJmH	sH	h�h��CJaJmH	sH	h�hz�CJaJmH	sH	h�htSCJaJmH	sH	h�hNw�CJaJmH	sH	h�h�g!CJaJmH	sH	h�hfZCJaJmH	sH	h�h�:�CJaJmH	sH	�~�~�~QXjkl����R�Y���'�.�́΁
��y������������у؃Y�Z���ф��b�i�j�4�5�q����Ƕ��ӘӌӘӌӘӌӘӁy�m�ӌӘӌӘӁm�ӌ�h�$�h�$�6�^JaJh4�^JaJh�$�h�$�^JaJh4�^JaJmH	sH	h�$�h�$�6�^JaJmH	sH	h�u�^JmH	nH
,sH	tH
, h�u�h�$�^JmH	nH
,sH	tH
,h�$�^JaJmH	sH	h�$�h�$�^JaJmH	sH	jh�h�CJUaJh�$�h�c�CJaJmH	sH	*����[���6�N�B���Ɉ�������?�@�B�L�����
�(�����������������������$d�$7$8$H$Ifa$gd�B�d�7$8$H$gd�
����0�d�^��`�0�gd4�d�gd4�q�x�L�M�����"�@�A�LjȈ,�3�4�������������������=�>�@�A�B�C�L�����������ɵ��������������क�tc h�h�
�CJ^JaJmH	sH	#h�h�
�5�CJ^JaJmH	sH	h�
�h�
�CJaJmH	sH	h�ZYh�
�CJaJmH	sH	jh�h�CJUaJUh�$�h�$�6�^JaJh4�^JaJh�$�h�$�^JaJh4�^JaJmH	sH	h�$�h�$�^JaJmH	sH	h�$�h�$�6�^JaJmH	sH	!14.	Emmerzaal, A, de Wit, JJ, Dijkstra, T, Bakker, D van Zijderveld, FG (2002) The Dutch Brucella abortus monitoring programme for cattle: the impact of false-positive serological reactions and comparison of serological tests. Vet Q 24: 40-46.
15.	Stuart, FACorbel, MJ (1982) Identification of a serological cross-reaction between Brucella abortus and Escherichia coli 0:157. Vet Rec 110: 202-203.
16.	Weynants, V, Tibor, A, Denoel, PA, Saegerman, C, Godfroid, J et al. (1996) Infection of cattle with Yersinia enterocolitica O:9 a cause of the false positive serological reactions in bovine brucellosis diagnostic tests. Vet Microbiol 48: 101-112.
17.	Samartino, L, Gregoret, R, Gall, D Nielsen, K (1999) Fluorescence polarization assay: application to the diagnosis of bovine brucellosis in Argentina. J Immunoassay 20: 115-126.


Table 1 
Criteria for interpretation of results from conventional agglutination tests in female cattle vaccinated between 3 and 8 month of age in Argentina.

BPASATb�-MEInterpretationNegativeNegativePositived" 50 IUNegativeNegativePositived" 25 IUd" 25 IUNegativePositive25 to 50 IUI25 IUNegativePositiveI100 IU to I200 IUNegativeSuspectedPositivee" I50 IUe" I50 IUPositivePositive200 IUNegativePositive
IU: international units
Table 2

Comparative performance of the fluorescence polarization assay for the detection of antibodies against the O-polysaccharide of B. abortus using different dilutions of the sera and antigen
		
Serum 1:100
Antigen 1:100PositiveNegativeSerum 1:10
Antigen 1:20Positive200Negative3115
Table 3

Comparative performance of fluorescence polarization assay with conventional agglutination tests in serum samples from an endemic region of Argentina

�������������(�*�:�<�R�T�d�f�������������,�.�>�@�����������������ᩍ��p\�p\�p\�p\�p\�p\�p&h�h�
�5�CJ\�^JaJmH	sH	#h�h�
�CJ\�^JaJmH	sH	h�h�
�CJaJ7h�h�
�5�B*CJOJQJ\�^JaJmH	phsH	/h�h�
�5�B*CJ\�^JaJmH	phsH	 h�
�h�
�CJ^JaJmH	sH	h�h�
�CJaJmH	sH	 h�h�
�CJ^JaJmH	sH	h�
�CJ^JaJmH	sH	$(�*�<�+$d�$7$8$H$Ifa$gd�B��kd�$$If�l��\���|a�"������������	�
t��(�����������������0���6��������������������4�4�
la�p�(����������������yt�B�<�>�@�R�T�f����0��kd_�$$If�l��\���|a�"����������������������������
t��0���6������������������������������������4�4�
la�yt�B�$d�$7$8$H$Ifa$gd�B�$
&Fd�$7$8$H$Ifa$gd�B�f�v����������������G�����kd[�$$If�l��\���|a�"��������������������������������������������
t��0���6����������������������������������������������������4�4�
la�yt�B�$d�$7$8$H$Ifa$gd�B������,�[GGGG$d�$7$8$H$Ifa$gd�B��kdW�$$If�l��\���|a�"��������������������������������������������
t��0���6����������������������������������������������������4�4�
la�yt�B�,�.�@�f�x���[GGGG$d�$7$8$H$Ifa$gd�B��kdS�$$If�l��\���|a�"��������������������������������������������
t��0���6����������������������������������������������������4�4�
la�yt�B�����������[GGGG$d�$7$8$H$Ifa$gd�B��kdO�$$If�l��\���|a�"��������������������������������������������
t��0���6����������������������������������������������������4�4�
la�yt�B��������[GGGG$d�$7$8$H$Ifa$gd�B��kdK�$$If�l��\���|a�"��������������������������������������������
t��0���6����������������������������������������������������4�4�
la�yt�B����� �(�)�1�Q�R�^�l������
���"�#�:������������~�h]hJ]h%h�hq�B*CJaJmH	phsH	h�hq�CJaJ+h�hq�5�B*CJ\�aJmH	phsH	h�hq�6�CJaJmH	sH	hq�CJaJmH	sH	h�hq�CJaJmH	sH	h�hq�5�CJaJmH	sH	hq�mH	sH	h�h�
�CJaJmH	sH	h�h�
�CJaJ h�h�
�CJ^JaJmH	sH	&h�h�
�5�CJ\�^JaJmH	sH	���(�1�2��[OOG;;d�7$8$H$gdq�d�gdq�d�7$8$H$gd�
��kdG�$$If�l��\���|a�"����������������������������
t��0���6������������������������������������4�4�
la�yt�B�������
������$d�$7$8$H$Ifa$gd�B�d�7$8$H$gdq�
�����"�J6666$d�$7$8$H$Ifa$gd�B��kdC�$$If�l��F��
�����	�
t���������������0��������������������4�4�
la�Qp�������������yt�B�"�#�.�,$d�$7$8$H$Ifa$gd�B��kdL�$$If�l��\��
�������	�
t��(�����������������0���������������������������4�4�
la�Qp�(����������������yt�B�.�;�D�G�I�����$d�$7$8$H$Ifa$gd�B�:�;�I�J�K�Z�[�]�e�����������������$
$$J$K$W$X$]$^$������²���������t_M_M_t"h!85�B*CJaJmH	phsH	(h�h!85�B*CJaJmH	phsH	h�h!8CJaJU+h�h!85�B*CJ\�aJmH	phsH	h!8CJaJmH	sH	h�h!8CJaJmH	sH	h�h!85�CJaJmH	sH	h!8mH	sH	h�hq�CJaJ%h�hq�B*CJaJmH	phsH	.h�hq�5�6�B*CJ\�aJmH	phsH	I�J�K�+d�$7$8$H$Ifgd�B��kdW�$$If�l4��\��
���`����	�
t��(�����������������0���������������������������4�4�
la�Qp�(����������������yt�B�K�T�V�Z����$d�$7$8$H$Ifa$gd�B�Z�[�\�+#d�gd!8�kd\�$$If�l4��\��
��� ����	�
t��(���������������0���������������������������4�4�
la�Qp�(����������������������yt�B�\�e�f�������
$$������@ykda�$$If�l��0r�3Y���������&��������
t �0��������������������������4�4�
la��yt�B�$d�$7$8$H$Ifa$gd�B�d�$7$8$H$Ifgd�B�d�7$8$H$gd!8d�7$8$H$gd!8d�gd!8Result ofNumber of samplesConventional agglutination testsFluorescent polarization assay27PositivePositive1PositiveSuspected9PositiveNegative9SuspectedNegative67NegativeNegative








 PAGE   \* MERGEFORMAT 3




$$>$]$^$���9�kd�$$If�l��Fr�3FY�������������������������	�
t ��������������0����������������������������������������4�4�
la��p�������������yt�B�$d�$7$8$H$Ifa$gd�B�^$a$j$s$t$v$$�$���\����kd�$$If�l��Fr�3FY�
t �0����������������������4�4�
la��yt�B�$d��x$7$8$H$Ifa$gd�B�^$s$t$�$�$�$�$�$�$�$�$�$�$�$�$�$�$�$�$�$�$�$�$�$�$�$�$�$�$�������������������¾µ¾���h�'�h-
mHnHuh�Njh�NUh�_cjh�_cUh2zqh/C�mH	sH	h�h!8CJaJ%h�h!8B*CJaJmH	phsH	�$�$�$�$�$M777$d��x$7$8$H$Ifa$gd�B��kd��$$If�l��Fr�3FY�������������������������	�
t ��������������0����������������������������������������4�4�
la��p�������������yt�B��$�$�$�$�$r\\\$d��x$7$8$H$Ifa$gd�B��kd��$$If�l��Fr�3FY�
t �0����������������������4�4�
la��yt�B��$�$�$�$�$M777$d��x$7$8$H$Ifa$gd�B��kd8�$$If�l��Fr�3FY�������������������������	�
t ��������������0����������������������������������������4�4�
la��p�������������yt�B��$�$�$�$�$�$�$�$�$�$�$rjb`b`b`b`d�gd�Nd�gd!8�kd.�$$If�l��Fr�3FY�
t �0����������������������4�4�
la��yt�B�
�$�$�$�$�$�$�����d�gd!8(($a$90P1�h:p�N��/ ��=!��"��#��$��%��������D<EndNote><Cite><Author>Schurig</Author><Year>1991</Year><RecNum>128</RecNum><record><rec-number>128</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">128</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Schurig, G. G.</author><author>Roop, R. M., 2nd</author><author>Bagchi, T.</author><author>Boyle, S.</author><author>Buhrman, D.</author><author>Sriranganathan, N.</author></authors></contributors><auth-address>Department of Pathobiology, Virginia-Maryland Regional College of Veterinary Medicine, Blacksburg, VA 24061.</auth-address><titles><title>Biological properties of RB51; a stable rough strain of Brucella abortus</title><secondary-title>Vet Microbiol</secondary-title></titles><periodical><full-title>Vet Microbiol</full-title></periodical><pages>171-88</pages><volume>28</volume><number>2</number><edition>1991/07/01</edition><keywords><keyword>Animals</keyword><keyword>Antigens, Bacterial/immunology</keyword><keyword>Blotting, Western</keyword><keyword>Brucella abortus/drug effects/*physiology</keyword><keyword>Brucellosis/microbiology</keyword><keyword>Brucellosis, Bovine/microbiology</keyword><keyword>Cattle</keyword><keyword>Drug Resistance, Microbial</keyword><keyword>Goats</keyword><keyword>Mice</keyword><keyword>Mice, Inbred BALB C</keyword><keyword>Rabbits</keyword><keyword>Rifampin/pharmacology</keyword></keywords><dates><year>1991</year><pub-dates><date>Jul</date></pub-dates></dates><isbn>0378-1135 (Print)&#xD;0378-1135 (Linking)</isbn><accession-num>1908158</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=1908158</url></related-urls></urls><electronic-resource-num>0378-1135(91)90091-S [pii]</electronic-resource-num><language>eng</language></record></Cite><Cite><Author>Nielsen</Author><Year>1996</Year><RecNum>40</RecNum><record><rec-number>40</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">40</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Nielsen, K.</author><author>Smith, P.</author><author>Gall, D.</author><author>Perez, B.</author><author>Cosma, C.</author><author>Mueller, P.</author><author>Trottier, J.</author><author>Cote, G.</author><author>Boag, L.</author><author>Bosse, J.</author></authors></contributors><auth-address>Agriculture and Agri-Food Canada, Animal Diseases Research Institute, Nepean, Ontario, Canada.</auth-address><titles><title>Development and validation of an indirect enzyme immunoassay for detection of antibody to Brucella abortus in milk</title><secondary-title>Vet Microbiol</secondary-title></titles><periodical><full-title>Vet Microbiol</full-title></periodical><pages>165-73</pages><volume>52</volume><number>1-2</number><edition>1996/09/01</edition><keywords><keyword>Animals</keyword><keyword>Antibodies, Bacterial/*analysis</keyword><keyword>Antibodies, Monoclonal</keyword><keyword>Antibody Specificity</keyword><keyword>Brucella abortus/*immunology/isolation &amp; purification</keyword><keyword>Brucellosis, Bovine/*diagnosis/*immunology</keyword><keyword>Cattle</keyword><keyword>Enzyme-Linked Immunosorbent Assay/methods/veterinary</keyword><keyword>Epitopes/analysis</keyword><keyword>Female</keyword><keyword>Immunoglobulin G</keyword><keyword>Lipopolysaccharides/immunology</keyword><keyword>Mice</keyword><keyword>Milk/*immunology/*microbiology</keyword><keyword>Reproducibility of Results</keyword><keyword>Sensitivity and Specificity</keyword></keywords><dates><year>1996</year><pub-dates><date>Sep</date></pub-dates></dates><isbn>0378-1135 (Print)&#xD;0378-1135 (Linking)</isbn><accession-num>8914260</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=8914260</url></related-urls></urls><language>eng</language></record></Cite></EndNote>�D<EndNote><Cite><Author>Nielsen</Author><Year>2004</Year><RecNum>66</RecNum><record><rec-number>66</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">66</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Nielsen, K.</author><author>Smith, P.</author><author>Widdison, J.</author><author>Gall, D.</author><author>Kelly, L.</author><author>Kelly, W.</author><author>Nicoletti, P.</author></authors></contributors><auth-address>Animal Diseases Research Institute, Canadian Food Inspection Agency, P.O. Box 11300, 3851 Fallowfield Road, Nepean, Ontario, Canada K2H 8P9. nielsenk@inspection.gc.ca</auth-address><titles><title>Serological relationship between cattle exposed to Brucella abortus, Yersinia enterocolitica O:9 and Escherichia coli O157:H7</title><secondary-title>Vet Microbiol</secondary-title></titles><periodical><full-title>Vet Microbiol</full-title></periodical><pages>25-30</pages><volume>100</volume><number>1-2</number><edition>2004/05/12</edition><keywords><keyword>Animals</keyword><keyword>Antibodies, Bacterial/*blood</keyword><keyword>Brucella abortus</keyword><keyword>Brucellosis, Bovine/*blood/diagnosis/microbiology</keyword><keyword>Cattle</keyword><keyword>Cross Reactions</keyword><keyword>Enzyme-Linked Immunosorbent Assay/veterinary</keyword><keyword>Escherichia coli/growth &amp; development</keyword><keyword>Escherichia coli Infections/blood/diagnosis/microbiology/*veterinary</keyword><keyword>Fluorescence Polarization Immunoassay/veterinary</keyword><keyword>Yersinia Infections/blood/diagnosis/microbiology/*veterinary</keyword><keyword>Yersinia enterocolitica/growth &amp; development</keyword></keywords><dates><year>2004</year><pub-dates><date>May 20</date></pub-dates></dates><isbn>0378-1135 (Print)&#xD;0378-1135 (Linking)</isbn><accession-num>15135510</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=15135510</url></related-urls></urls><electronic-resource-num>10.1016/j.vetmic.2003.12.010&#xD;S0378113504000513 [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>�D<EndNote><Cite><Author>Aguirre</Author><Year>2002</Year><RecNum>67</RecNum><record><rec-number>67</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">67</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Aguirre, N. P.</author><author>Vanzini, V. R.</author><author>Torioni de Echaide, S.</author><author>Valentini, B. S.</author><author>De Lucca, G.</author><author>Aufranc, C.</author><author>Canal, A.</author><author>Vigliocco, A.</author><author>Nielsen, K.</author></authors></contributors><auth-address>Instituto Nacional de Tecnologia Agropecuaria, E. E. A. Rafaela, CC 22, CP 2300, Rafaela (Santa Fe), Argentina.</auth-address><titles><title>Antibody dynamics in Holstein Friesian heifers vaccinated with Brucella abortus strain 19, using seven serological tests</title><secondary-title>J Immunoassay Immunochem</secondary-title></titles><periodical><full-title>J Immunoassay Immunochem</full-title></periodical><pages>471-8</pages><volume>23</volume><number>4</number><edition>2002/12/03</edition><keywords><keyword>Animals</keyword><keyword>Brucella Vaccine/*immunology</keyword><keyword>Brucella abortus/*immunology</keyword><keyword>Brucellosis, Bovine/prevention &amp; control</keyword><keyword>Cattle</keyword><keyword>Complement Fixation Tests</keyword><keyword>Enzyme-Linked Immunosorbent Assay/methods</keyword><keyword>Immunoglobulin Isotypes/*biosynthesis</keyword><keyword>Time Factors</keyword><keyword>Vaccines</keyword></keywords><dates><year>2002</year></dates><isbn>1532-1819 (Print)&#xD;1532-1819 (Linking)</isbn><accession-num>12458730</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=12458730</url></related-urls></urls><language>eng</language></record></Cite><Cite><Author>Nielsen</Author><Year>1998</Year><RecNum>33</RecNum><record><rec-number>33</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">33</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Nielsen, K.</author><author>Gall, D.</author><author>Lin, M.</author><author>Massangill, C.</author><author>Samartino, L.</author><author>Perez, B.</author><author>Coats, M.</author><author>Hennager, S.</author><author>Dajer, A.</author><author>Nicoletti, P.</author><author>Thomas, F.</author></authors></contributors><auth-address>Canadian Food Inspection Agency, Animal Diseases Research Institute, Nepean, Ont., Canada. nielsenk@em.agr.ca</auth-address><titles><title>Diagnosis of bovine brucellosis using a homogeneous fluorescence polarization assay</title><secondary-title>Vet Immunol Immunopathol</secondary-title></titles><periodical><full-title>Vet Immunol Immunopathol</full-title></periodical><pages>321-9</pages><volume>66</volume><number>3-4</number><edition>1999/01/08</edition><keywords><keyword>Animals</keyword><keyword>Brucella abortus/immunology</keyword><keyword>Brucellosis, Bovine/*diagnosis/prevention &amp; control</keyword><keyword>Cattle</keyword><keyword>Fluorescence Polarization/methods/*veterinary</keyword><keyword>Sensitivity and Specificity</keyword><keyword>Software</keyword></keywords><dates><year>1998</year><pub-dates><date>Dec 11</date></pub-dates></dates><isbn>0165-2427 (Print)&#xD;0165-2427 (Linking)</isbn><accession-num>9880108</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=9880108</url></related-urls></urls><electronic-resource-num>S0165-2427(98)00195-0 [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>~D<EndNote><Cite><Author>Ramirez-Pfeiffer</Author><Year>2008</Year><RecNum>13</RecNum><record><rec-number>13</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">13</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Ramirez-Pfeiffer, C.</author><author>Diaz-Aparicio, E.</author><author>Rodriguez-Padilla, C.</author><author>Morales-Loredo, A.</author><author>Alvarez-Ojeda, G.</author><author>Gomez-Flores, R.</author></authors></contributors><auth-address>Instituto Nacional de Investigaciones Forestales y Agropecuarias, Campo Experimental Rio Bravo, Rio Bravo, Tamaulipas, Mexico.</auth-address><titles><title>Improved performance of Brucella melitensis native hapten over Brucella abortus OPS tracer on goat antibody detection by the fluorescence polarization assay</title><secondary-title>Vet Immunol Immunopathol</secondary-title></titles><periodical><full-title>Vet Immunol Immunopathol</full-title></periodical><pages>223-9</pages><volume>123</volume><number>3-4</number><edition>2008/03/25</edition><keywords><keyword>Animals</keyword><keyword>Antibodies, Bacterial/*blood/immunology</keyword><keyword>Brucella abortus/immunology</keyword><keyword>Brucella melitensis/chemistry/*immunology</keyword><keyword>Brucellosis/diagnosis/immunology/microbiology/*veterinary</keyword><keyword>Cattle</keyword><keyword>Fluorescein-5-isothiocyanate/chemistry</keyword><keyword>Fluorescence Polarization Immunoassay/methods/*veterinary</keyword><keyword>Fluorescent Dyes/chemistry</keyword><keyword>Goat Diseases/blood/diagnosis/immunology/*microbiology</keyword><keyword>Goats</keyword><keyword>Haptens/*chemistry/immunology</keyword><keyword>ROC Curve</keyword><keyword>Reproducibility of Results</keyword><keyword>Sensitivity and Specificity</keyword></keywords><dates><year>2008</year><pub-dates><date>Jun 15</date></pub-dates></dates><isbn>0165-2427 (Print)&#xD;0165-2427 (Linking)</isbn><accession-num>18359093</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=18359093</url></related-urls></urls><electronic-resource-num>S0165-2427(08)00062-7 [pii]&#xD;10.1016/j.vetimm.2008.02.001</electronic-resource-num><language>eng</language></record></Cite><Cite><Author>Minas</Author><Year>2007</Year><RecNum>15</RecNum><record><rec-number>15</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">15</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Minas, A.</author><author>Stournara, A.</author><author>Minas, M.</author><author>Stack, J.</author><author>Petridou, E.</author><author>Christodoulopoulos, G.</author><author>Krikelis, V.</author></authors></contributors><auth-address>Technological Educational Institution of Larissa, Faculty of Health Professions, Laboratory of Microbiology, Larissa, 411 10, Greece. aminas@teilar.gr</auth-address><titles><title>Validation of a fluorescence polarization assay (FPA) performed in microplates and comparison with other tests used for diagnosing B. melitensis infection in sheep and goats</title><secondary-title>J Immunol Methods</secondary-title></titles><periodical><full-title>J Immunol Methods</full-title></periodical><pages>94-103</pages><volume>320</volume><number>1-2</number><edition>2007/01/30</edition><keywords><keyword>Animals</keyword><keyword>Brucella melitensis/*immunology</keyword><keyword>Brucellosis/blood/*diagnosis/veterinary</keyword><keyword>Fluorescence Polarization Immunoassay/*methods/standards/veterinary</keyword><keyword>Goat Diseases/blood/*diagnosis</keyword><keyword>Goats</keyword><keyword>ROC Curve</keyword><keyword>Reference Values</keyword><keyword>Reproducibility of Results</keyword><keyword>Sheep Diseases/blood/*diagnosis</keyword><keyword>Sheep, Domestic</keyword></keywords><dates><year>2007</year><pub-dates><date>Mar 30</date></pub-dates></dates><isbn>0022-1759 (Print)&#xD;0022-1759 (Linking)</isbn><accession-num>17258229</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=17258229</url></related-urls></urls><electronic-resource-num>S0022-1759(06)00374-7 [pii]&#xD;10.1016/j.jim.2006.12.008</electronic-resource-num><language>eng</language></record></Cite><Cite><Author>McGiven</Author><Year>2003</Year><RecNum>44</RecNum><record><rec-number>44</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">44</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>McGiven, J. A.</author><author>Tucker, J. D.</author><author>Perrett, L. L.</author><author>Stack, J. A.</author><author>Brew, S. D.</author><author>MacMillan, A. P.</author></authors></contributors><auth-address>Department of Bacterial Diseases, FAO/WHO Collaborating Centre for Reference and Research on Brucellosis, Veterinary Laboratories Agency, Woodham Lane, New Haw, Weybridge, Surrey, KT15 3NB, UK. j.mcgiven@vla.defra.gsi.gov.uk</auth-address><titles><title>Validation of FPA and cELISA for the detection of antibodies to Brucella abortus in cattle sera and comparison to SAT, CFT, and iELISA</title><secondary-title>J Immunol Methods</secondary-title></titles><periodical><full-title>J Immunol Methods</full-title></periodical><pages>171-8</pages><volume>278</volume><number>1-2</number><edition>2003/09/06</edition><keywords><keyword>Agglutination Tests/veterinary</keyword><keyword>Animals</keyword><keyword>Brucella abortus/*immunology</keyword><keyword>Brucellosis, Bovine/diagnosis/*immunology</keyword><keyword>Cattle</keyword><keyword>Complement Fixation Tests/veterinary</keyword><keyword>Enzyme-Linked Immunosorbent Assay/*veterinary</keyword><keyword>Fluorescence Polarization Immunoassay/*veterinary</keyword><keyword>Reproducibility of Results</keyword><keyword>Sensitivity and Specificity</keyword></keywords><dates><year>2003</year><pub-dates><date>Jul</date></pub-dates></dates><isbn>0022-1759 (Print)&#xD;0022-1759 (Linking)</isbn><accession-num>12957405</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=12957405</url></related-urls></urls><electronic-resource-num>S0022175903002011 [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>b	D<EndNote><Cite><Author>Ramirez-Pfeiffer</Author><Year>2008</Year><RecNum>13</RecNum><record><rec-number>13</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">13</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Ramirez-Pfeiffer, C.</author><author>Diaz-Aparicio, E.</author><author>Rodriguez-Padilla, C.</author><author>Morales-Loredo, A.</author><author>Alvarez-Ojeda, G.</author><author>Gomez-Flores, R.</author></authors></contributors><auth-address>Instituto Nacional de Investigaciones Forestales y Agropecuarias, Campo Experimental Rio Bravo, Rio Bravo, Tamaulipas, Mexico.</auth-address><titles><title>Improved performance of Brucella melitensis native hapten over Brucella abortus OPS tracer on goat antibody detection by the fluorescence polarization assay</title><secondary-title>Vet Immunol Immunopathol</secondary-title></titles><periodical><full-title>Vet Immunol Immunopathol</full-title></periodical><pages>223-9</pages><volume>123</volume><number>3-4</number><edition>2008/03/25</edition><keywords><keyword>Animals</keyword><keyword>Antibodies, Bacterial/*blood/immunology</keyword><keyword>Brucella abortus/immunology</keyword><keyword>Brucella melitensis/chemistry/*immunology</keyword><keyword>Brucellosis/diagnosis/immunology/microbiology/*veterinary</keyword><keyword>Cattle</keyword><keyword>Fluorescein-5-isothiocyanate/chemistry</keyword><keyword>Fluorescence Polarization Immunoassay/methods/*veterinary</keyword><keyword>Fluorescent Dyes/chemistry</keyword><keyword>Goat Diseases/blood/diagnosis/immunology/*microbiology</keyword><keyword>Goats</keyword><keyword>Haptens/*chemistry/immunology</keyword><keyword>ROC Curve</keyword><keyword>Reproducibility of Results</keyword><keyword>Sensitivity and Specificity</keyword></keywords><dates><year>2008</year><pub-dates><date>Jun 15</date></pub-dates></dates><isbn>0165-2427 (Print)&#xD;0165-2427 (Linking)</isbn><accession-num>18359093</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=18359093</url></related-urls></urls><electronic-resource-num>S0165-2427(08)00062-7 [pii]&#xD;10.1016/j.vetimm.2008.02.001</electronic-resource-num><language>eng</language></record></Cite></EndNote>rD<EndNote><Cite><Author>Minas</Author><Year>2007</Year><RecNum>15</RecNum><record><rec-number>15</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">15</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Minas, A.</author><author>Stournara, A.</author><author>Minas, M.</author><author>Stack, J.</author><author>Petridou, E.</author><author>Christodoulopoulos, G.</author><author>Krikelis, V.</author></authors></contributors><auth-address>Technological Educational Institution of Larissa, Faculty of Health Professions, Laboratory of Microbiology, Larissa, 411 10, Greece. aminas@teilar.gr</auth-address><titles><title>Validation of a fluorescence polarization assay (FPA) performed in microplates and comparison with other tests used for diagnosing B. melitensis infection in sheep and goats</title><secondary-title>J Immunol Methods</secondary-title></titles><periodical><full-title>J Immunol Methods</full-title></periodical><pages>94-103</pages><volume>320</volume><number>1-2</number><edition>2007/01/30</edition><keywords><keyword>Animals</keyword><keyword>Brucella melitensis/*immunology</keyword><keyword>Brucellosis/blood/*diagnosis/veterinary</keyword><keyword>Fluorescence Polarization Immunoassay/*methods/standards/veterinary</keyword><keyword>Goat Diseases/blood/*diagnosis</keyword><keyword>Goats</keyword><keyword>ROC Curve</keyword><keyword>Reference Values</keyword><keyword>Reproducibility of Results</keyword><keyword>Sheep Diseases/blood/*diagnosis</keyword><keyword>Sheep, Domestic</keyword></keywords><dates><year>2007</year><pub-dates><date>Mar 30</date></pub-dates></dates><isbn>0022-1759 (Print)&#xD;0022-1759 (Linking)</isbn><accession-num>17258229</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=17258229</url></related-urls></urls><electronic-resource-num>S0022-1759(06)00374-7 [pii]&#xD;10.1016/j.jim.2006.12.008</electronic-resource-num><language>eng</language></record></Cite></EndNote>ZD<EndNote><Cite><Author>McGiven</Author><Year>2003</Year><RecNum>44</RecNum><record><rec-number>44</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">44</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>McGiven, J. A.</author><author>Tucker, J. D.</author><author>Perrett, L. L.</author><author>Stack, J. A.</author><author>Brew, S. D.</author><author>MacMillan, A. P.</author></authors></contributors><auth-address>Department of Bacterial Diseases, FAO/WHO Collaborating Centre for Reference and Research on Brucellosis, Veterinary Laboratories Agency, Woodham Lane, New Haw, Weybridge, Surrey, KT15 3NB, UK. j.mcgiven@vla.defra.gsi.gov.uk</auth-address><titles><title>Validation of FPA and cELISA for the detection of antibodies to Brucella abortus in cattle sera and comparison to SAT, CFT, and iELISA</title><secondary-title>J Immunol Methods</secondary-title></titles><periodical><full-title>J Immunol Methods</full-title></periodical><pages>171-8</pages><volume>278</volume><number>1-2</number><edition>2003/09/06</edition><keywords><keyword>Agglutination Tests/veterinary</keyword><keyword>Animals</keyword><keyword>Brucella abortus/*immunology</keyword><keyword>Brucellosis, Bovine/diagnosis/*immunology</keyword><keyword>Cattle</keyword><keyword>Complement Fixation Tests/veterinary</keyword><keyword>Enzyme-Linked Immunosorbent Assay/*veterinary</keyword><keyword>Fluorescence Polarization Immunoassay/*veterinary</keyword><keyword>Reproducibility of Results</keyword><keyword>Sensitivity and Specificity</keyword></keywords><dates><year>2003</year><pub-dates><date>Jul</date></pub-dates></dates><isbn>0022-1759 (Print)&#xD;0022-1759 (Linking)</isbn><accession-num>12957405</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=12957405</url></related-urls></urls><electronic-resource-num>S0022175903002011 [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>ZD<EndNote><Cite><Author>McGiven</Author><Year>2003</Year><RecNum>44</RecNum><record><rec-number>44</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">44</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>McGiven, J. A.</author><author>Tucker, J. D.</author><author>Perrett, L. L.</author><author>Stack, J. A.</author><author>Brew, S. D.</author><author>MacMillan, A. P.</author></authors></contributors><auth-address>Department of Bacterial Diseases, FAO/WHO Collaborating Centre for Reference and Research on Brucellosis, Veterinary Laboratories Agency, Woodham Lane, New Haw, Weybridge, Surrey, KT15 3NB, UK. j.mcgiven@vla.defra.gsi.gov.uk</auth-address><titles><title>Validation of FPA and cELISA for the detection of antibodies to Brucella abortus in cattle sera and comparison to SAT, CFT, and iELISA</title><secondary-title>J Immunol Methods</secondary-title></titles><periodical><full-title>J Immunol Methods</full-title></periodical><pages>171-8</pages><volume>278</volume><number>1-2</number><edition>2003/09/06</edition><keywords><keyword>Agglutination Tests/veterinary</keyword><keyword>Animals</keyword><keyword>Brucella abortus/*immunology</keyword><keyword>Brucellosis, Bovine/diagnosis/*immunology</keyword><keyword>Cattle</keyword><keyword>Complement Fixation Tests/veterinary</keyword><keyword>Enzyme-Linked Immunosorbent Assay/*veterinary</keyword><keyword>Fluorescence Polarization Immunoassay/*veterinary</keyword><keyword>Reproducibility of Results</keyword><keyword>Sensitivity and Specificity</keyword></keywords><dates><year>2003</year><pub-dates><date>Jul</date></pub-dates></dates><isbn>0022-1759 (Print)&#xD;0022-1759 (Linking)</isbn><accession-num>12957405</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=12957405</url></related-urls></urls><electronic-resource-num>S0022175903002011 [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>�D<EndNote><Cite><Author>Minas</Author><Year>2007</Year><RecNum>15</RecNum><record><rec-number>15</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">15</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Minas, A.</author><author>Stournara, A.</author><author>Minas, M.</author><author>Stack, J.</author><author>Petridou, E.</author><author>Christodoulopoulos, G.</author><author>Krikelis, V.</author></authors></contributors><auth-address>Technological Educational Institution of Larissa, Faculty of Health Professions, Laboratory of Microbiology, Larissa, 411 10, Greece. aminas@teilar.gr</auth-address><titles><title>Validation of a fluorescence polarization assay (FPA) performed in microplates and comparison with other tests used for diagnosing B. melitensis infection in sheep and goats</title><secondary-title>J Immunol Methods</secondary-title></titles><periodical><full-title>J Immunol Methods</full-title></periodical><pages>94-103</pages><volume>320</volume><number>1-2</number><edition>2007/01/30</edition><keywords><keyword>Animals</keyword><keyword>Brucella melitensis/*immunology</keyword><keyword>Brucellosis/blood/*diagnosis/veterinary</keyword><keyword>Fluorescence Polarization Immunoassay/*methods/standards/veterinary</keyword><keyword>Goat Diseases/blood/*diagnosis</keyword><keyword>Goats</keyword><keyword>ROC Curve</keyword><keyword>Reference Values</keyword><keyword>Reproducibility of Results</keyword><keyword>Sheep Diseases/blood/*diagnosis</keyword><keyword>Sheep, Domestic</keyword></keywords><dates><year>2007</year><pub-dates><date>Mar 30</date></pub-dates></dates><isbn>0022-1759 (Print)&#xD;0022-1759 (Linking)</isbn><accession-num>17258229</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=17258229</url></related-urls></urls><electronic-resource-num>S0022-1759(06)00374-7 [pii]&#xD;10.1016/j.jim.2006.12.008</electronic-resource-num><language>eng</language></record></Cite><Cite><Author>Ramirez-Pfeiffer</Author><Year>2008</Year><RecNum>7</RecNum><record><rec-number>7</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">7</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Ramirez-Pfeiffer, C.</author><author>Diaz-Aparicio, E.</author><author>Gomez-Flores, R.</author><author>Rodriguez-Padilla, C.</author><author>Morales-Loredo, A.</author><author>Alvarez-Ojeda, G.</author></authors></contributors><auth-address>Campo Experimental Rio Bravo, Instituto Nacional de Investigaciones Forestales y Agricolas y Pecuarias, Rio Bravo, Tam., Mexico.</auth-address><titles><title>Use of the Brucella melitensis native hapten to diagnose brucellosis in goats by a rapid, simple, and specific fluorescence polarization assay</title><secondary-title>Clin Vaccine Immunol</secondary-title></titles><periodical><full-title>Clin Vaccine Immunol</full-title></periodical><pages>911-5</pages><volume>15</volume><number>6</number><edition>2008/04/04</edition><keywords><keyword>Animals</keyword><keyword>Antibodies, Bacterial/*blood</keyword><keyword>Antigens, Bacterial/immunology</keyword><keyword>Brucella melitensis/*isolation &amp; purification</keyword><keyword>Brucellosis/*microbiology</keyword><keyword>Cattle</keyword><keyword>Enzyme-Linked Immunosorbent Assay/veterinary</keyword><keyword>Fluorescence Polarization Immunoassay/methods/*veterinary</keyword><keyword>Goat Diseases/*diagnosis/immunology</keyword><keyword>Goats</keyword><keyword>Haptens/*immunology</keyword><keyword>Sensitivity and Specificity</keyword></keywords><dates><year>2008</year><pub-dates><date>Jun</date></pub-dates></dates><isbn>1556-679X (Electronic)&#xD;1556-679X (Linking)</isbn><accession-num>18385457</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=18385457</url></related-urls></urls><electronic-resource-num>CVI.00046-08 [pii]&#xD;10.1128/CVI.00046-08</electronic-resource-num><language>eng</language></record></Cite></EndNote>�D<EndNote><Cite><Author>Nielsen</Author><Year>1996</Year><RecNum>1</RecNum><record><rec-number>1</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">1</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Nielsen, K.</author><author>Gall, D.</author><author>Jolley, M.</author><author>Leishman, G.</author><author>Balsevicius, S.</author><author>Smith, P.</author><author>Nicoletti, P.</author><author>Thomas, F.</author></authors></contributors><auth-address>Agriculture and Agri-Food Canada, Animal Diseases Research Institute, Nepean, Ont., Canada.</auth-address><titles><title>A homogeneous fluorescence polarization assay for detection of antibody to Brucella abortus</title><secondary-title>J Immunol Methods</secondary-title></titles><periodical><full-title>J Immunol Methods</full-title></periodical><pages>161-8</pages><volume>195</volume><number>1-2</number><edition>1996/09/09</edition><keywords><keyword>Animals</keyword><keyword>Antibodies, Bacterial/*blood</keyword><keyword>Brucella abortus/*immunology</keyword><keyword>Cattle</keyword><keyword>*Fluorescence Polarization Immunoassay</keyword></keywords><dates><year>1996</year><pub-dates><date>Sep 9</date></pub-dates></dates><isbn>0022-1759 (Print)&#xD;0022-1759 (Linking)</isbn><accession-num>8814332</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=8814332</url></related-urls></urls><language>eng</language></record></Cite><Cite><Author>Nielsen</Author><Year>1998</Year><RecNum>33</RecNum><record><rec-number>33</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">33</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Nielsen, K.</author><author>Gall, D.</author><author>Lin, M.</author><author>Massangill, C.</author><author>Samartino, L.</author><author>Perez, B.</author><author>Coats, M.</author><author>Hennager, S.</author><author>Dajer, A.</author><author>Nicoletti, P.</author><author>Thomas, F.</author></authors></contributors><auth-address>Canadian Food Inspection Agency, Animal Diseases Research Institute, Nepean, Ont., Canada. nielsenk@em.agr.ca</auth-address><titles><title>Diagnosis of bovine brucellosis using a homogeneous fluorescence polarization assay</title><secondary-title>Vet Immunol Immunopathol</secondary-title></titles><periodical><full-title>Vet Immunol Immunopathol</full-title></periodical><pages>321-9</pages><volume>66</volume><number>3-4</number><edition>1999/01/08</edition><keywords><keyword>Animals</keyword><keyword>Brucella abortus/immunology</keyword><keyword>Brucellosis, Bovine/*diagnosis/prevention &amp; control</keyword><keyword>Cattle</keyword><keyword>Fluorescence Polarization/methods/*veterinary</keyword><keyword>Sensitivity and Specificity</keyword><keyword>Software</keyword></keywords><dates><year>1998</year><pub-dates><date>Dec 11</date></pub-dates></dates><isbn>0165-2427 (Print)&#xD;0165-2427 (Linking)</isbn><accession-num>9880108</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=9880108</url></related-urls></urls><electronic-resource-num>S0165-2427(98)00195-0 [pii]</electronic-resource-num><language>eng</language></record></Cite><Cite><Author>Aguirre</Author><Year>2002</Year><RecNum>67</RecNum><record><rec-number>67</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">67</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Aguirre, N. P.</author><author>Vanzini, V. R.</author><author>Torioni de Echaide, S.</author><author>Valentini, B. S.</author><author>De Lucca, G.</author><author>Aufranc, C.</author><author>Canal, A.</author><author>Vigliocco, A.</author><author>Nielsen, K.</author></authors></contributors><auth-address>Instituto Nacional de Tecnologia Agropecuaria, E. E. A. Rafaela, CC 22, CP 2300, Rafaela (Santa Fe), Argentina.</auth-address><titles><title>Antibody dynamics in Holstein Friesian heifers vaccinated with Brucella abortus strain 19, using seven serological tests</title><secondary-title>J Immunoassay Immunochem</secondary-title></titles><periodical><full-title>J Immunoassay Immunochem</full-title></periodical><pages>471-8</pages><volume>23</volume><number>4</number><edition>2002/12/03</edition><keywords><keyword>Animals</keyword><keyword>Brucella Vaccine/*immunology</keyword><keyword>Brucella abortus/*immunology</keyword><keyword>Brucellosis, Bovine/prevention &amp; control</keyword><keyword>Cattle</keyword><keyword>Complement Fixation Tests</keyword><keyword>Enzyme-Linked Immunosorbent Assay/methods</keyword><keyword>Immunoglobulin Isotypes/*biosynthesis</keyword><keyword>Time Factors</keyword><keyword>Vaccines</keyword></keywords><dates><year>2002</year></dates><isbn>1532-1819 (Print)&#xD;1532-1819 (Linking)</isbn><accession-num>12458730</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=12458730</url></related-urls></urls><language>eng</language></record></Cite></EndNote>�D<EndNote><Cite><Author>Stuart</Author><Year>1982</Year><RecNum>70</RecNum><record><rec-number>70</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">70</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Stuart, F. A.</author><author>Corbel, M. J.</author></authors></contributors><titles><title>Identification of a serological cross-reaction between Brucella abortus and Escherichia coli 0:157</title><secondary-title>Vet Rec</secondary-title></titles><periodical><full-title>Vet Rec</full-title></periodical><pages>202-3</pages><volume>110</volume><number>9</number><edition>1982/02/27</edition><keywords><keyword>Animals</keyword><keyword>Antigens, Bacterial/immunology</keyword><keyword>Brucella abortus/*immunology</keyword><keyword>Cattle</keyword><keyword>Cross Reactions</keyword><keyword>Escherichia coli/*immunology</keyword><keyword>Immune Sera/immunology</keyword><keyword>Rabbits</keyword></keywords><dates><year>1982</year><pub-dates><date>Feb 27</date></pub-dates></dates><isbn>0042-4900 (Print)&#xD;0042-4900 (Linking)</isbn><accession-num>6803435</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=6803435</url></related-urls></urls><language>eng</language></record></Cite><Cite><Author>Weynants</Author><Year>1996</Year><RecNum>71</RecNum><record><rec-number>71</rec-number><foreign-keys><key app="EN" db-id="9wx0xfzeizzp26evsxkpsweztrrzwxd0wd0d">71</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Weynants, V.</author><author>Tibor, A.</author><author>Denoel, P. A.</author><author>Saegerman, C.</author><author>Godfroid, J.</author><author>Thiange, P.</author><author>Letesson, J. J.</author></authors></contributors><auth-address>Unite d&apos;Immunologie-Microbiologie, Universitaires Notre-Dame de la Paix, Namur, Belgium.</auth-address><titles><title>Infection of cattle with Yersinia enterocolitica O:9 a cause of the false positive serological reactions in bovine brucellosis diagnostic tests</title><secondary-title>Vet Microbiol</secondary-title></titles><periodical><full-title>Vet Microbiol</full-title></periodical><pages>101-12</pages><volume>48</volume><number>1-2</number><edition>1996/01/01</edition><keywords><keyword>Animals</keyword><keyword>Antibodies, Bacterial/*blood</keyword><keyword>Antigens, Bacterial</keyword><keyword>Bacterial Outer Membrane Proteins</keyword><keyword>Brucella abortus/*immunology</keyword><keyword>Brucellosis, Bovine/*diagnosis/microbiology</keyword><keyword>Cattle</keyword><keyword>Enzyme-Linked Immunosorbent Assay/methods/*veterinary</keyword><keyword>False Positive Reactions</keyword><keyword>Female</keyword><keyword>Yersinia enterocolitica/*immunology</keyword></keywords><dates><year>1996</year><pub-dates><date>Jan</date></pub-dates></dates><isbn>0378-1135 (Print)&#xD;0378-1135 (Linking)</isbn><accession-num>8701566</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=8701566</url></related-urls></urls><language>eng</language></record></Cite></EndNote>M$$If�!vh5��5��5��5��#v�#v�#v�#v�:V�l	�
t��(�����������������0���6�5��5��5��5��/���������/�/���������p�(����������������yt�B��$$If�!vh5��5��5��5��#v�#v�#v�#v�:V�l
t��0���6�5��5��5��5��/���������/�/���������/���������yt�B��$$If�!vh5��5��5��5��#v�#v�#v�#v�:V�l
t��0���6�5��5��5��5��/���������/�
��������/���������/���������yt�B��$$If�!vh5��5��5��5��#v�#v�#v�#v�:V�l
t��0���6�5��5��5��5��/���������/�
��������/���������/���������yt�B��$$If�!vh5��5��5��5��#v�#v�#v�#v�:V�l
t��0���6�5��5��5��5��/���������/�
��������/���������/���������yt�B��$$If�!vh5��5��5��5��#v�#v�#v�#v�:V�l
t��0���6�5��5��5��5��/���������/�
��������/���������/���������yt�B��$$If�!vh5��5��5��5��#v�#v�#v�#v�:V�l
t��0���6�5��5��5��5��/���������/�
��������/���������/���������yt�B��$$If�!vh5��5��5��5��#v�#v�#v�#v�:V�l
t��0���6�5��5��5��5��/���������/�	��������/���������/�yt�B�$$If�Q!vh5��5��5��#v�#v�:V�l	�
t���������������0������5��5��/�/��a�Qp�������������yt�B�	$$If�Q!vh5��5��5��5��#v�:V�l	�
t��(�����������������0������5��/��a�Qp�(����������������yt�B�$$If�Q!vh5��5��5��5��#v�:V�l4	�
t��(�����������������0������+�5��a�Qp�(����������������yt�B�$$If�Q!vh5��5��5��5��#v�:V�l4	�
t��(���������������0������+�5��a�Qp�(����������������������yt�B��$$If��!vh5��5�&#v�#v&:V�l
t �0�����5��5�&/�/�
�������a��yt�B��$$If��!vh5��5�5�#v�#v:V�l	�
t ��������������0�����5��5�/�
�������a��p�������������yt�B��$$If��!vh5��5�5�#v�#v:V�l
t �0�����5��5�a��yt�B��$$If��!vh5��5�5�#v�#v:V�l	�
t ��������������0�����5��5�/�
�������a��p�������������yt�B��$$If��!vh5��5�5�#v�#v:V�l
t �0�����5��5�a��yt�B��$$If��!vh5��5�5�#v�#v:V�l	�
t ��������������0�����5��5�/�
�������a��p�������������yt�B��$$If��!vh5��5�5�#v�#v:V�l
t �0�����5��5�a��yt�B�j+���������666666666vvvvvvvvv666666>666666666666666666666666666�6666666666�666666666666hH66666666666666666666666666666666666666666666666666666666666666666�62���� 0@P`p������2(�� 0@P`p������ 0@P`p������ 0@P`p������ 0@P`p������ 0@P`p������ 0@P`p��8X�V~ OJPJQJ_HmH
,nH
,sH
,tH
,L`�L$Normal$da$CJ_HaJmH
,sH
,tH	bb�=��T�tulo 1$$��@&'5B*CJOJPJQJ\^JaJph6_�bb�=��T�tulo 2$$��@&'5B*CJOJPJQJ\^JaJphO��ZZ�=��T�tulo 3$$��@&5B*OJPJQJ\^JphO��NA`�N
Fuente de p�rrafo predeter.Ri@�R0Tabla normal�4�
l4�a�,k �,
0	Sin listaD�@�D�?� P�rrafo de lista	^��m$h�o�h�9�Default$7$8$H$a$-B*CJOJQJ^J_HaJmH
,phsH
,tH	�����c��Tabla con cuadr�cula7:V�0d�z��#z�0Q�Lista clara1�:V�0���4�4j�.@�������j�.�������j�.�������j���
�.d�f�d���f�d���5��5\��5\��5\��5B*\ph���h^2h	xT�0Normal (Web)d��d�d[$\$a$CJOJPJQJ^JaJtH
,<U@�A<xT�0Hiperv�nculo>*B*ph�X��QX�=��T�tulo 1 Car'5�B*CJOJPJQJ\�^JaJph6_�X��aX�=��T�tulo 2 Car'5�B*CJOJPJQJ\�^JaJphO��P��qP�=��T�tulo 3 Car5�B*OJPJQJ\�^JphO��FB�F�=�0Texto independiente�xF���F�=�0Texto independiente CarRC�R�=�0Sangr�a de texto normal
�x^�N���N�=�0Sangr�a de texto normal CarrN��r�=�0%Texto independiente primera sangr�a 2�^�h`�hj���j�=�0)Texto independiente primera sangr�a 2 Car��������Sombreado claroG:V�0�����4�4j�;@��������������������������������������
����j�;��������������������������������������
����j�D���������������������������������������j�D���������������������������������������(f�d���f�d���5B*ph��5\��5\��5\��5\D'��D
�<70Ref. de comentarioCJaJDD
!�<70Texto comentario CJaJD��D �<70Texto comentario CartH	LjL
#�<70Asunto del comentario"5�\�P�1P"�<70Asunto del comentario Car5\R�BR
%�<70Texto de globo$d�CJOJQJ^JaJT��QT$�<70Texto de globo CarCJOJQJ^JaJtH	<b<
'�N0
Encabezado
&
�C�"@��q@&�N0Encabezado CarCJaJtH	B @�B)�N0
Pie de p�gina
(
�C�"F���F(�N0Pie de p�gina CarCJaJtH	6(��6
�N0N�mero de l�neaPK!����[Content_Types].xml���j�0E����ж�r�(��΢Iw},��-j��4	��w�P�-t#bΙ{U�����T�U^h�d}㨫��)��*1P�'��	�^��W��0)��T�9<�l�#��$yi}��;�~@��(��H����u�*Dנz��/0�ǰ��$��X��3aZ����,�D0j~�3߶�b��~i>���3�\`�?�/�[���G��\�!�-�Rk.�s�Ի�..���a濭?��PK!�֧�6_rels/.rels���j�0���}Q��%v/��C/�}�(h"���O�
����=������ ����C?�h�v=��Ʌ��%[xp��{۵_�Pѣ<�1�H�0���O�R�Bd���JE�4b$��q_����6L�R�7`������0̞O��,�En7�Li�b��/�S���e��е�����PK!ky���theme/theme/themeManager.xml�M
� @�}�w��7c�(Eb�ˮ��C�AǠҟ����7��՛K
Y,�
�e�.���|,���H�,l����xɴ��I�sQ}#Ր���� ֵ+�!�,�^�$j=�GW��)�E�+&
8���PK!&lT�Rtheme/theme/theme1.xml�YMoE�#�F{oc'vGu�رH�F�[��xw�;���jf��7���q�7��J\ʯ	A��xgfw��I�FPA}H����1^�3tH��<i{��5���M¶w{ؿ��!�p`���Dz�6��*^W�	�D��)��/-I����S���11V�(¥@�#����Zmu)�4�P�c`;tk<�>�6r�=2%����@3'M	�5BNe�	t�Y�I?���CK/�^�|����Kx=#bjm��o>]F,�"B�F��V�����z�n�^�3�`�ե̳�_�wr�%��:ϻ[k�.��eN�V��i�2],S�_s���jcs���7���f����
��W��+�Ն�7����`���g�Ș��J���j|��l(�K��D-ʵ��
dX��iJ�؇<��x$(��:��7vɗsKZ����j{�jb�����_=��<;~���Ç�~���m��e���~����O�y��j�,���_~���3S�ŗO~{���W���ݣ
����2|Hc"�Mr��y������8�0´L���'XK���S���9�,���G���#�}T�O�9
"1Q�B�N;�]�Y��J/�hY%7'IX-\Lʸ}��dwq�ķ7I�o�i�ލ���É�!I�B�? �º��:~ݥ���KQ�J���ɦ�6�!.�*�!ގov�gUVo�C	U�Y��C�7^���*�C���o`U)9�
���I�	�)�hn	����2�l�H��A���2r�t#�U�M�2�y)��WU�]�V�~�8�da��P��np���J��o&B�Z�Ӂc��];f��́�k��_|��"���F�	{RU%l�h��p'�n�����=wO�=i>��k��Z���o����v�[����fD�N�c��@M�!͐,a�������'�4��Y_wp���	�>�*D8���i&��X��\��,W��xҕ=6������.��^���ۄ��Z��*l�J��~au�ԙ�Սj��9�
�!��b�M@�-��U8�k�p0����v���b�p�!���9�7���1�� �bn w*b�y�x�$��پ����,��@\�7�R���(�=Q�,)'K�Q�k5���q��p���q
Q�z��,��!_	������Y4[�an���}�`��B�-,#��U�,ђ���Mp�E`3�5�XY�d�״?��%�1�U9إ�;���R>QD����D�c�NU�'��&LG�p���m^��9+����u��g�V�h^�n��<���*u7Ɲ�S�dJ9��g��n
V.rF�^�*�Ѕ҈�}����-p�!��:��P��5gy����ڧ!�#	B��-��;�Y=ۻ,K�12URW�V�9$l�{���=A��n���;��sVA�P9�zszH�����'[�`�ۇ�@���P�bW��<�{ˆ��1��W+m���_S�sn��c�Y��̕�(�[��@��}�`���g��	���>�V?4hf�6�՗��t���#��M&�ʺ6������'�B�	gk���s:��\qN-^��3;��k]
�=Y��4�2&0�G��N|t�����I&�MI`=����DC����PK!
ѐ��'theme/theme/_rels/themeManager.xml.rels��M
�0���wooӺ�&݈Э���5
6?$Q��
�,.�a��i����c2�1h�:�q��m��@RN��;d�`��o7�g�K(M&$R(.1�r'J��ЊT���8��V�"��AȻ�H�u}��|�$�b{��P����8�g/]�QAsم(����#��L�[������PK-!����[Content_Types].xmlPK-!�֧�6+_rels/.relsPK-!ky���theme/theme/themeManager.xmlPK-!&lT�R�theme/theme/theme1.xmlPK-!
ѐ��'�	theme/theme/_rels/themeManager.xml.relsPK]�
<?xml version="1.0" encoding="UTF-8" standalone="yes"?>
<a:clrMap xmlns:a="http://schemas.openxmlformats.org/drawingml/2006/main" bg1="lt1" tx1="dk1" bg2="lt2" tx2="dk2" accent1="accent1" accent2="accent2" accent3="accent3" accent4="accent4" accent5="accent5" accent6="accent6" hlink="hlink" folHlink="folHlink"/>�6����	***-��
���^�g�� <!F#�$�,.-j.�1(3_4�6�8:�;�=�>�@�D�GOI�LNeO�R3V�WwYJ[A]�^g3h�i5rV|�~q����:�^$�$EGHIJKMNOPQRSTUVWXYZ[\]^_abcdefghijkmnopqrstuvwy~�����8�V��(�<�f��,������
�"�.�I�K�Z�\�$^$�$�$�$�$�$�$FL`lx�����������������������]aGWmnt&<=A�� � 6"F"\"]"c"�#&&T(d(z({(�(44+4,4046�8�8r;�;�;"@2@H@I@N@y@�@�@�@�@K�K�K�K�KLL*L+L3L(O8ONOOOUOeP�W�W�Z(b-b�b�b�b�b�b9c�i�i�m�mI|�Q���QQ�@���QQ�@���Q���QQ�@���Q���QQ�@���QQ�@����Q����X���QQ�@����QQ�@����QQ�@����QQ�@����QQ�@����Q����Q����QQ�@���Q����Q����$&-!����L�#��A�A@�����������0�(	�
��B
�S����	?���_GoBack
�
�AIJQ������&+hk�����������
�
8B��7?�����
�
p|��
/>08ls��� !!! !.![!b!("0"''�'�'�(�(�)�)J,Q,�,�,'-.-F-Q-R-`-0.;.N.[./"/�/�/�2�2�2�2�2�2�2�2�2�2�2�2�2�2,3.3�3�3�4�4�4�4�4�4�4�4�4�455551535O5Q5997:>:�:�:�:;%;-;N;V;)<4<�<�<�>�>k@r@�B�BVHdHkHrH�H�H�I�I�J�JbKmK�K�K�W�W�X�X�b�b�b�b�b�b�b�b)c2cj-jnjuj^kik�l�l�l�l�l�l�l�l�l�l�m�m�m�m�m�mn
nYncndninonrn�n�n�n�n�n�n�n�n
oooo#o,ofoko�o�o�o�o�o�o%p-p�p�p�p�p�p�p�p�p�p�pq	qqqq&q�q�q�q�q�q�q�q�q�q�q�q�q�q�q�q�qQrYrZrar�r�r�r�r�r�rUs\s]sist.tEtNt�t�t�t�t�t�t�t�t�t�t/u6u7uCu`uiu�u�u�u�u"v,vMvTvnvuv�v�v�v�v�v�v�vw6w<w@wGwHwOwjwqw�w�wxx!x*xbxjxkxux}x�x�x�x�x�x
yy'y/yByLybyjykyry~y�y	zzTz\z]zdz�z�z�z�z�z�z�z�z�z�z�z�z�z{{{{{�{�{�{�{�{�~�~��������������€ĀŀǀȀ��������qs��`"b"�.�.�0�0�1�1�2�2�2�2�2�2�B�BC!C/L2LiLqL�s�s�s�s�t*u�x�x{{��������������€ĀŀǀȀ��3333333333333333333333�^"f"�*�*�;�;�G�G�m�mYn�no:o�op�p�p�q�q�r�rQsws+uRuKvjv>w^w�w�w�x	y�y�y�z�zw{�{-|L|�|$~�~X�����������������€ĀŀǀȀ���A%>t�}����������<�]�3@"���������08qH�^���������������^��`���o(.������^��`���.��p�L�^�p`�L�.��@���^�@`���.�����^�`���.����L�^��`�L�.������^��`���.������^��`���.��P�L�^�P`�L�.������^��`���OJPJQJ^Jo(-��e���^�e`���OJQJ^Jo(o��5���^�5`���OJQJo(����
���^�
`���OJQJo(��������^��`���OJQJ^Jo(o������^��`���OJQJo(����u���^�u`���OJQJo(����E���^�E`���OJQJ^Jo(o�����^�`���OJQJo(�������^��`���5o(.�
�����^��`����h�H.�
�p�L�^�p`�L��h�H.�
�@���^�@`����h�H.�
����^�`����h�H.�
���L�^��`�L��h�H.�
�����^��`����h�H.�
�����^��`����h�H.�
�P�L�^�P`�L��h�H.�A%>�<�]08q����������������
,
,
,
,
,
,
,
,
,D���
,
,
,
,
,
,
,
,
,
,
,
,
,
,
,
,
,�_�w�&�D����jtH�h�k��:�^(�(�[�a%�.8�JHh�x-
�6
7A
�5�PfZ,�R�v`���wB@�LMAr�@�y�L�M�N�o�
*pX�k��b$�kd�6P�
 x!�"!_!�g!�"U'"�J"OK"�<#$
	$�y$�%�R%�J&"M("c)>*�-*mM*�o*bx*�,A0,IP,8-�%-f..�].W1�1�'1�2�;23�h4�05�H5�W6�7�97�<7�`7!8s{8EO9�:�$:�%:`d:�<�)<=u<�	>�5>-p>�
?X2?�+AcB�B�'BzIB�.Ca_C8D�
EDF�H�DH�LMLqM�9N�OoP�,P�GP�0Q8Q�qQ�RWR�'RLnStS�hT�jT�rVS2W�=W�X�Y�ZY�pY�wY}Z�#[W\�?\P\r>]�O]"2_�b#Yc�_c1|c�<d�kewYg%h�gh1ilYj/k`(k�4klJk�Ok$l(*m	n10nfn�oPyo�p)Ap2zq�,sKrs�*t%ytuvb-w�Rw+yTy�	z�M{-U{�p|�}}2}{O}mQ~�Z~�g~�>.:0ko�Fx�z����p��~��[��`�:��p�s�2���9�=��E�"?�t �wg�
�f.��
����S�kc��m��R�b[��i�4���7�}���X�:�+?�O��P�D��^����4�E�g�P�M"��*��F�[^��(��E�����2#�UW�ca�s�fv��}��k�t�Hk��&��h�U��:�hW�A�^ ��E��x�{��k�K�.s�>�1�1o�O�	e�Xt�k���+��=��c�Nw������B���D*�g�(Z����9>�a3���)A��o��@�T�,��@��B�8E��_���'�62�%@��b��$�yz�����[��e��A�v��[�5O�t�z������L��h����8��o�xT��i�"Z��y��C�L��n�+p�L��z��z��8�6H��F���C^��v��9�/C����?���4m����m�~8��K��E��
��c��g�sG�����D-��J�W}�)�����n<�UI�RT��u��o��*�d�uk��Q��	���0?��B��d�Ig�q��1���b�������EN.InstantFormat��	EN.Layout�EN.Libraries�xw<ENInstantFormat><Enabled>1</Enabled><ScanUnformatted>1</ScanUnformatted><ScanChanges>1</ScanChanges></ENInstantFormat>R<ENLayout><Style>JVetSciMedDiag</Style><LeftDelim>{</LeftDelim><RightDelim>}</RightDelim><FontName>Calibri</FontName><FontSize>11</FontSize><ReflistTitle></ReflistTitle><StartingRefnum>1</StartingRefnum><FirstLineIndent>0</FirstLineIndent><HangingIndent>720</HangingIndent><LineSpacing>0</LineSpacing><SpaceAfter>0</SpaceAfter></ENLayout>N<ENLibraries><Libraries><item>Brucellosis.enl</item></Libraries></ENLibraries>�@���--
5
6�9�:�C�Djk	y�|�}��X@XX,@X XD@X@X�@XFX�@XRX�@XzX�@X�@X�X�@XH@��Unknown������������	G��*�Ax�	�Times New Roman5��Symbol3.��*�Cx�	�Arial5.��.�[`�)�Tahoma7.���@	�Calibri7���K@�Cambria?=�	�*�Cx�	�Courier New;��WingdingsA���� B�Cambria Math"1�����#g�#�._��lA����lA��$���n���0{�{�>�Q���HX	�$P����������������������t�2!xx���UsuarioUsuario����Oh��+'��0T�������	��

(4<DL�UsuarioNormalUsuario46Microsoft Office Word@��1@�
�C�@��E���l����՜.��+,��D��՜.��+,��8�hp��������
���Hewlett-Packard�A{�T�tulo� 8@_PID_HLINKS�A�U[%-http://www.graphpad.com/quickcalcs/index.cfm&	

 !"#$%&'()*+,-./0123456789:;<=>?@ABCDEFGHIJKLMNOPQRSTUVWXYZ[\]^_`abcdefghijklmnopqrstuvwxyz{|}~��������������������������������������������������������������������������������������������������������������������������������������	

�������� !"#$%����������������*+��������.����������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������Root Entry��������	�FK�E�-Data
�������������˧1Table���������wJWordDocument
����;6SummaryInformation(������������DocumentSummaryInformation8��������MsoDataStore����������%�E�pBG�E��H����X�YE�PYD�Q0JD5�A==2����������%�E�pBG�E�Item
����	�����Properties������������UCompObj����
}����������������	����������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������<b:Sources SelectedStyle="\APA.XSL" StyleName="APA" xmlns:b="http://schemas.openxmlformats.org/officeDocument/2006/bibliography" xmlns="http://schemas.openxmlformats.org/officeDocument/2006/bibliography"></b:Sources><?xml version="1.0" encoding="UTF-8" standalone="no"?>
<ds:datastoreItem ds:itemID="{B3E97B94-E9D5-4860-8F60-38D06890DFD8}" xmlns:ds="http://schemas.openxmlformats.org/officeDocument/2006/customXml"><ds:schemaRefs><ds:schemaRef ds:uri="http://schemas.openxmlformats.org/officeDocument/2006/bibliography"/></ds:schemaRefs></ds:datastoreItem>��
����	�F+Documento de Microsoft Office Word 97-2003
MSWordDocWord.Document.8�9�q

Youez - 2016 - github.com/yon3zu
LinuXploit