The Comparison of Rumen Fungi Quantification in the Medium Containing Sunflower Meal Treated with Formaldehyde and or Sodium Hydroxide by Using Quantitative Competitive PCR

Abstract

The method of quantitative competitive polymerase chain reaction (QC-PCR) was conducted to compare the number of rumen anaerobic fungi in pure culture of fungi containing high fat sunflower meal (165 g fat/kg DM) processed with formaldehyde and NaOH. Twenty one multiparous early lactating Holstein cows (30±5 days of lactation) selected and fed experimental diets for 7 weaks. The diets were including untreated sunflower meal (control, n=3) and treated 4 % sodium hydroxide (n=3) and treated with 0.3 and 0.6 % formaldehyde (n=3). Competitive PCR technique was used to evaluate quantitative difference of anaerobic fungal population in the rumen under the dietary treatments. Standard control DNA was constructed from lambda phage for use in the competitive PCR and was shown to amplify under the same reaction condition and with the same amplification efficiency as the target DNA. The relative intensities of PCR products were used to evaluate variety of fungal population under fed treatments. The analysis of data of present study showed that NaOH treated sunflower meal increased and formaldehyde treated sunflower meal decreased number of fungi in medium compared to control. Therefore it seems that QC-PCR method has appropriate efficacy for enumerating rumen fungal population under the effect of dietary treatments.