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Enzyme-linkedi mmunosorbenta ssay  (ELISA). The ELISAu sed in this study has been describedp reviously (20, 26).  Briefly, 96-well flat-bottom plates (Immuno-plate Maxisorp, Nunc, Denmark) were coated with rabbit antiserumi n carbonate buffer (pH 9.6) and incubated at 4 C overnight.  The plates were then washed with phosphateb uffer (pH 7.4) containing 0 .05% Tween 20 and blockedw ith 1 % bovine serum albumini n carbonate buffer at 37 C for 1 hr. After washing the plates, six serial 2-fold dilutions of the intestinal contents were made with  a  phosphate  buffer  (pH  7.4)  containing   1%  bovine serum  albumin  and  0.05%  Tween  20.  Aliquots  (100   IA) of  the  dilutions  were  added  to  each  well  and  incubated  at
37  C  for  45   min.  Detection  was  carried  out  with  sheep anti-mouse  IgG  conjugated  with  horseradish  peroxidase (Caltag  Laboratories,  San  Francisco,  Calif.,  U.S.A.), using  o-phenylene  diamine  (Wako  Pure  Chemical  Industries,  Tokyo)  as  the  substrate.  The  plates  were  read with  an  EIA  reader   (Model   2550,  Bio-Rad  Laboratories, Richmond,  Calif,  U.S.A.)  at  492  nm.  The  number  of CBM588  vegetative  cells  were  determined  by  interpolation  from  a  standard  curve.
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