Table 3.

Enterococcal DNA in Cecal Fluids

Feeding trial	Group	Ct	Log10 counts	Fold increase
103–4 weeks	Water	26.14	5.69	–
	Ethanol	21.76	7.27	38 (47)
114–14 weeks	Water	33.67	2.98	–
	Ethanol	25.97	5.75	587

Cecal fluid of C57BL/6 mice was carefully harvested and diluted for cultures as described in Materials and Methods, after 4 or 14 weeks of 20% ethanol feeding. Aliquots of the diluted fluid were processed for PCR detection and quantitation of DNA for Escherichia coli β-glucosidase and Enterococcus 23S rRNA, exactly as described by Frahm and Obst, 2003. Standard curves of log₁₀ of bacterial number in control cultures processed for PCR versus C_t were constructed; the slopes of the curves were essentially identical to the control curves published by Frahm and Obst. C_t represents the threshold cycle in which the fluorescence signal exceeds background. There was a dramatic increase in Enterococcus DNA in ethanol mice (38 to over 500-fold in separate determinations), consistent with the culture results. The parenthetic result shown for fold increase in trial 103–4 represents a separate PCR determination of another sample from the same feeding trial. E. coli was isolated in very few colonies by culture, and specific DNA was only marginally increased in the cecal fluid of ethanol mice as compared with water controls (data not shown).