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Effect of leaves of six forest plants on rumen microbiota and fermentation characteristics in vitro.

D. P. Bu



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Effect of leaves of six forest plants on rumen microbiota and fermentation characteristics in vitro.
A. Ayemele Gnetegha1, L. Ma1, T. Park2, J. C. Xu1,3, Z. T. Yu2, D. P. Bu*1,4. 1State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences Beijing, China, 2Department of Animal Sciences, The Ohio State University Columbus, OH, 3World Agroforestry Center, East and Central Asia Kunming, China, 4CAASICRAF Joint Lab on Agroforestry and Sustainable Animal Husbandry, World Agroforestry Centre, East and Central Asia Beijing, China.

This study screened the leaves of 6 forest plants (Brassica rapa chinensis, Kalimeris indica, Calotropis procera, Portulara oleracea, Flemingia macrophylla, and Adansonia digitata) at 4 different doses (0, 0.7, 0.9 and 1.1 mg/mL) for their ability to inhibit rumen protozoa in vitro. Rumen fluid was collected from 3 different dairy cows and cultured anaerobically for 24 h incubation in the presence of ground leaves of each plant and then counted under a light microscope. The protozoal cells were examined using scanning electron microscopy for potential changes in cell surface morphology. Total bacteria and total archaea were quantified as copies of 16S rRNA genes per mL culture samples using qPCR. The VFA and ammonia concentration were determined using gas chromatography and colorimetry respectively. Data were analyzed in a completely randomized design using the PROC GLM procedure of SAS 9.4. Out of the 6 tested plants, only Calotropis procera inhibited the genus Entodinium, which is the main culprit of intraruminal microbial protein cycling, and decreased ammonia nitrogen production (P < 0.05), while not suppressing the cellulolytic genera Orphryoscolex or Eudiplodinium (P > 0.05). The leaves of C. Procera damaged the Entodinium extracellular structure in a dose-dependent manner. Total bacterial and archaeal populations were not decreased (P > 0.05). Except for butyrate which increased (P < 0.05), the molar proportion of VFAs and total VFAs production were not altered (P > 0.05). Chemical extraction and analysis of the C. procera leaves revealed phenolics (48.98 � 1.89 mg/g of dry extract), flavonoids (8.93 � 0.89 mg/g), and alkaloids (3.61 � 0.75 mg/g) as the major secondary metabolites. Future research is warranted to identify the specific secondary metabolite(s) that are responsible for the specific inhibition of Entodinium and the underpinning inhibition mechanisms.

Keywords: anaerobic culture, nitrogen efficiency, Calotropis procera.