Effects of using different roughages in the total mixed ration inoculated with or without coculture of Lactobacillus acidophilus and Bacillus subtilis on in vitro rumen fermentation and microbial population

Abstract

Objective: This study aimed to determine the effects of different roughages in total mixed ration (TMR) inoculated with or without coculture of <i>Lactobacillus acidophilus</i> (<i>L. acidophilus</i>) and <i>Bacillus subtilis</i> (<i>B. subtilis</i>) on <i>in vitro</i> rumen fermentation and microbial population.Methods: Three TMRs formulations composed of different forages were used and each TMR was grouped into two treatments: non-fermented TMR and fermented TMR (F-TMR) (inoculated with coculture of <i>L. acidophilus</i> and <i>B. subtilis</i>). After fermentation, the fermentation, chemical and microbial profile of the TMRs were determined. The treatments were used for <i>in vitro</i> rumen fermentation to determine total gas production, pH, ammonianitrogen (NH<sub>3</sub>-N), and volatile fatty acids (VFA). Microbial populations were determined by quantitative real-time polymerase chain reaction (PCR). All data were analyzed as a 3×2 factorial arrangement design using the MIXED procedure of Statistical Analysis Systems.Results: Changes in the fermentation (pH, lactate, acetate, propionate, and NH<sub>3</sub>-N) and chemical composition (moisture, crude protein, crude fiber, and ash) were observed. For <i>in vitro</i> rumen fermentation, lower rumen pH, higher acetate, propionate, and total VFA content were observed in the F-TMR group after 24 h incubation (p<0.05). F-TMR group had higher acetate concentration compared with the non-fermented group. Total VFA was highest (p<0.05) in F-TMR containing combined forage of domestic and imported source (F-CF) and F-TMR containing Italian ryegrass silage and corn silage (F-IRS-CS) than that of TMR diet containing oat, timothy, and alfalfa hay. The microbial population was not affected by the different TMR diets.Conclusion: The use of Italian ryegrass silage and corn silage, as well as the inoculation of coculture of <i>L. acidophilus</i> and <i>B. subtilis</i>, in the TMR caused changes in the pH, lactate and acetate concentrations, and chemical composition of experimental diets. In addition, F-TMR composed with Italian ryegrass silage and corn silage altered ruminal pH and VFA concentrations during <i>in vitro</i> rumen fermentation experiment.

Kimchi cabbage (Brassica rapa L.) by-products treated with calcium oxide and alkaline hydrogen peroxide as feed ingredient for Holstein steers

“…The serum bottle (160 mL capacity) was prepared by adding 1 g of weighed substrate of each treatment with three replications. One hundred milliliter of buffered rumen fluid was filled into individual serum bottles containing the substrate, under anaerobic conditions [ 22 ]. The bottles were subsequently sealed with rubber septum stoppers and aluminum caps and incubated at 39°C for 6, 12, and 24 h with shaking (100 rpm) [ 23 ].…”

“…Samples of fermentation were also collected in 1.5 mL cryotubes and deep frozen at −80°C. These samples were later thawed at room temperature and then centrifuged at 13,000×g for 15 min at 4°C using a Micro 17TR centrifuge (Hanil Science Industrial, Incheon, Korea) and the supernatant was used for ammonia nitrogen (NH 3 -N) and volatile fatty acids (VFA) analysis [ 22 ]. Using the Libra S22 spectrophotometer (Biochrom, Cambridge, UK) at an absorbance of 630 nm, NH 3 -N concentration was measured according to the methods developed by Chaney and Marbach [ 25 ].…”

Seasonal Influence on Rumen Microbiota, Rumen Fermentation, and Enteric Methane Emissions of Holstein and Jersey Steers under the Same Total Mixed Ration

“…CH 4 is an indicator of dietary gross energy losses, and it has a negative environmental impact contributing to global warming [ 23 , 24 ]. Previous studies revealed that several factors, including diet, feed additives, host genetics, age, and physiological state affect the rumen microbiomes, rumen fermentation characteristics, and CH 4 production [ 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 ]. O’Hara et al [ 28 ] reported an association between the rumen microbiome and its fermentation products with feed efficiency and CH 4 emissions.…”

Kecernaan, Fermentabilitas dan Produksi Protein Mikrobia Secara In Vitro pada Complete Feed Berbasis Pelepah Sawit Fermentasi

“…McDonald et al (2010) menyatakan bahwa, kisaran normal N-NH3 untuk menunjang pertumbuhan mikrobia rumen yang optimal yakni 6-21mM. Miguel et al (2021) melaporkan, pengaruh penggunaan serat yang berbeda dalam complete feed yang tidak fermentasi menghasilkan produksi N-NH3 sebesar 8,76 mM dan complete feed fermentasi 12,61 mM.…”