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Phytate content in feed ingredients can negatively impact digestibility and palatability. To address this issue, it is necessary to study microbes capable of breaking down phytate content. This study aimed to isolate and characterize phytase-producing bacteria from decaying materials rich in phytic acid. The research was conducted in several stages. The first stage involved isolating phytase-producing bacteria from the acidification of Tithonia diversifolia using growth media containing Na-phytate. Bacterial isolates that produced clear zones were then tested for their activity and ability to produce several enzymes, specifically phytase, cellulase, and protease. The next step was to test the morphological characteristics of the bacterial isolate. The final stage of bacterial identification consisted of DNA isolation, followed by PCR amplification of the 16S rRNA gene, DNA sequence homology analysis, and construction of a phylogenetic tree. Based on research, three isolates were found to produce clear phytase zones: isolates R5 (20.3 mm), R7 (16.1 mm) and R8 (31.7 mm). All isolates were able to produce the enzymes phytase (5.45-6.54 U/ml), cellulase (2.60-2.92 U/ml), and protease (22.2-23.4 U/ml). Metagenomic testing identified isolate R7 and R8 as Alcaligenes faecalis and isolate R5 as Achromobacter xylosoxidans. The isolation and characterization of phytase-producing bacteria from Tithonia diversifolia acidification resulted in the identification of two promising candidates that can be applied as sources of phytase producers. Phytase-producing bacteria can be utilized to improve digestibility and palatability in animal feed.
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Background and Aim: To achieve optimal feed efficiency in ruminants, especially Pesisir cattle, it is necessary to maintain a harmonious equilibrium between energy and protein levels within the rumen. Sulfur supplementation can potentially escalate the energy-protein balance in the rumen. The aim of this study was to explore the formulation of ruminant diets by synchronizing rumen degradable protein (RDP) and non-fiber carbohydrate (NFC) while adding sulfur minerals at different levels. Nutrient digestibility, NH3 concentration, volatile fatty acids (VFA) production, microbial protein synthesis (MPS), and methane gas production were assessed. Materials and Methods: We employed a randomized block design with a 2 × 2 × 3 factorial arrangement and examined diverse incubation periods of 6, 24, and 48 h. Treatment consisted of RDP (60% and 65%), NFC (35% and 40%), and sulfur (0%, 0.15%, and 0.3%) levels. In this study, the Tilley and Terry in vitro technique, which used Pesisir cattle's rumen fluid, was employed to assess the digestibility of dry matter, organic matter, acid detergent fiber, neutral detergent fiber, and RDP-Rumen undegradable protein. In addition, it measures various rumen fluid attributes, including pH, NH3, VFA, MPS, and methane gas production. Results: Treatment with a coordinated combination of 65% RDP and 40% NFC combined with 0.15% sulfur supplement yielded significantly improved digestibility and notably reduced methane gas production (p < 0.05). Conclusion: The enhancement in digestibility and reduction in methane gas emissions can be attributed to the interaction of RDP, NFC, and sulfur. Feed digestibility was increased in the 65% RDP treatment with 40% NFC and 0.15% sulfur, along with a decrease in methane gas production.
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Background and Aim: Probiotic supplementation can assist with manipulating the rumen microbial ecosystem. Lactic acid bacteria and yeast from fermented fish (Budu) as the indigenous food from West Sumatra, Indonesia, are potential probiotics for livestock. This study aims to select the best candidate lactic acid bacteria and yeast strains from fermented fish as ruminant probiotics and evaluate the effect of their supplementation on the characteristics of rumen fermentation, feed digestion, and total gas production in vitro. Materials and Methods: This study used nine treatments, performed in triplicate, in a completely randomized design. The substrate ratio comprised of 70% Pennisetum purpureum forage and 30% concentrate. Five lactic acid bacteria and three yeast isolates were used in this study. Treatments were as follows: T0: control (basal diet); T1: T0 + Lactobacillus parabuchneri strain 3347; T2: T0 + Lactobacillus buchneri strain 5296; T3: T0 + Lactobacillus harbinensis JCM 16178; T4: T0 + Schleiferilactobacillus harbinensis strain LH991; T5: T0 + L. parabuchneri strain 6902; T6: T0 + Pichia kudriavzevii strain B-5P; T7: T0 + P. kudriavzevii strain CBS 5147; and T8: T0 + commercial yeast (Saccharomyces cerevisiae). The lactic acid bacteria inoculum contained 1.02 × 1011 colony-forming unit (CFU)/mL, while the yeast inoculum contained 1.5 × 1010 CFU/mL. Results: The results showed that four lactic acid bacteria and three yeast produced a higher total gas yield (104-183.33 mL) compared to the control (103 mL). Supplementation with lactic acid bacteria in the rumen fermentation in vitro showed dry matter digestibility of 63%-70% and organic matter digestibility (OMD) of 64%-71%. We observed that total volatile fatty acid (VFA) production in all treatments was significantly higher (86-121 mM) compared to the control (81 mM). The concentration of NH3 production was higher in all treatments (12.33-16.83 mM) than in the control (12.25 mM). Meanwhile, the probiotic supplementation did not cause a significant change in the rumen pH (6.86-7.12). Supplementation with the lactic acid bacteria S. harbinensis strain LH991 consistently demonstrated the best results from the parameters of dry and OMD (70.29% and 71.16%, respectively), total VFA (121.67 mM), NH3 (16.83 mM), and total gas production (149.17 mL). The best results were observed from the yeast candidate P. kudriavzevii strain B-5P, where the results were dry and OMD (67.64% and 69.55% respectively), total VFA (96.67 mM), NH3 (13.42 mM), and total gas production (183.33 mL). Conclusion: Based on the obtained results, lactic acid bacteria S. harbinensis strain LH991 and yeast P. kudriavzevii strain B-5P are attractive candidates to be utilized as probiotics for ruminants based on their potential to improve rumen fermentation in vitro. This probiotic supplementation can increase the digestibility of feed ingredients, production of total VFA and NH3, and total gas produced.
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Objective: Protein and carbohydrates are substances needed by ruminants, especially sheep. Providing protein and carbohydrates must pay attention to their degradation. In addition, balancing nutrients to meet the nutritional needs of rumen microbes is very important because the unbalanced availability of rumen degradable protein (RDP) and readily available carbohydrate (RAC) at one time can cause suboptimal microbial protein synthesis efficiency. Materials and Methods: Completely randomized design with a nonfactorial pattern of five treatments with three replications. Treatment includes RDP and RAC ratios, namely R1 = 2.30, R2 = 2.00, R3 = 1.70, R4 = 1.50, and R5 = 1.30. Results: The results showed that the balance of RDP and RAC in sheep ration formulation in vitro had a very significant effect (p < 0.01) on NH3, microbial protein synthesis, total gas, total microbes, and organic matter digestibility (OMD) but had no significant effect (p > 0.05) on rumen pH and dry matter digestibility (DMD). Conclusion: The optimal balance of RDP and RAC in the formulation of sheep rations in vitro was obtained at a ratio of 2.30 with NH3 (mM) 8.47, rumen pH 5.97, microbial protein synthesis (mg/100 ml) 123, gas (ml/g of material) 145, total microbes (cells/ml) 2.012 × 106, (log CFU cells/ml) 6.3025, DMD 61.0%, and OMD 63.1%.
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Background: Tannins are a group of phenolic compounds that can modify the rumen biohydrogenation (BH) of polyunsaturated fatty acids (PUFA), but to date results obtained have been inconsistent. This study therefore aims to conduct a meta-analysis of the scientific literature related to the effects of tannins on rumen BH and fermentation. Methods: A total of 28 articles were collected from various scientific databases, such as Scopus, Science Direct and Google Scholar, and the data were analysed using a random effects model and meta-regression for rumen BH. The publication bias on the main variables of rumen fermentation was assessed using a funnel plot and Egger's test. Results: An increase in tannin levels significantly reduced methane production (p < 0.001) and the population of Butyrivibrio fibrisolvens (p < 0.05). Dietary tannins also decreased the SFA proportion (p < 0.001) and increased (p < 0.001) the rumen monounsaturated fatty acid (MUFA) and polyunsaturated fatty acid (PUFA) proportions. In additions, there were negative relationships between dietary tannin levels and BH rates of C18:2 n-6 and C18:3 n-3 (p < 0.05). Conclusion: Dietary tannins modulate the rumen fermentation profile, mitigate methane emissions, and inhibit rumen BH of PUFA.
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BACKGROUND AND AIM: Feeding ruminants must notice the degradability of feed, especially protein. Microbial rumen requires ammonia from rumen degradable protein (RDP) beside that ruminant require bypass protein or rumen undegradable protein (RUP) and microbial crude protein. The aim of the study was to discover the best RDP:RUP ratio in beef cattle diets commonly used by Indonesian farmers using an in vitro methodology. MATERIALS AND METHODS: Samples of Pennisetum purpureum, Leucaena leucocephala, Indigofera zollingeriana, cassava, maize, palm kernel cake, rice bran, and tofu waste were formulated into dietary treatments (dry matter [DM] basis). All experiments were carried out using a 3×3×2 factorial, randomized block design with three replications. Treatments consisted of three protein levels (12%, 14%, and 16%), two energy levels (65% and 70%), and three RDP:RUP ratio levels (55:45, 60:40, and 65:35). The experimental diets were incubated in vitro using buffered rumen fluid for 48 h at 39°C. After incubation, the supernatants were analyzed to determine pH, ammonia concentration, total volatile fatty acid (VFA), and microbial protein synthesis. The residues were analyzed to determine DM, organic matter, protein, and RUP digestibility. RESULTS: Increased protein, energy, and RDP levels increased digestibility, ammonia concentrations, total VFAs, and microbial protein synthesis (p<0.05), while rations with 16% protein lowered these parameters (p<0.05). CONCLUSION: Increased dietary protein (from 12% to 14% DM), energy (from 65% to 70% DM), and RDP (from 55% to 65% crude protein [CP]) levels increased nutrient digestibility, ammonia concentration, total VFA levels, and microbial protein synthesis. The diet containing 14% DM dietary protein and 70% DM energy, which contained 55%, 60%, or 65% CP RDP optimally increased nutrient digestibility, ammonia concentration, total VFA levels, and microbial protein synthesis. Thus, feed based on these RDP:RUP ratios can optimize ruminant productivity.
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AIM: This research aimed to discover the chemical composition, as well as the content of the degradable and undegradable protein of the ruminant feed commonly used as cattle feed by Indonesian farmers. MATERIALS AND METHODS: In this study, Pennisetum purpureum, Leucaena leucocephala, Indigofera zollingeriana, Gliricidia sepium, cassava, maize, palm kernel cake, and rice bran were used as feed. Chemical composition was determined by proximate and Van Soest analyses performed in triplicate. Dry matter and organic matter digestibility, as well as the rumen degradable proteins (RDP) and rumen undegradable proteins (RUP) contents, were determined in vitro using the Tilley and Terry method. RESULTS: The results showed that more proteins can be obtained from legumes than from grass or concentrates. The highest protein amount was obtained from I. zollingerian (31.22%), while the lowest protein amount was obtained from cassava (3.59%). Dry matter digestibility ranged from 18.53% (rice bran) to 49.21% (G. sepium). Organic matter digestibility ranged from 35.71% (cassava) to 59.57% (I. zollingerian). Rice bran had the highest RDP from concentrate (73.26%), whereas I. zollingerian had the highest RDP from forage (74.72%). The highest RUP from concentrate was obtained from palm kernel cake (61.01%), and the highest RUP from forage was obtained from L. leucocephala (49.23%). CONCLUSION: The preparation of ruminant livestock rations must be based on RDP and RUP to meet the needs of both the rumen microbes and host animals. Information regarding the RDP and RUP of feeds is still limited, making this study useful in the preparation of ruminant livestock rations based on RDP and RUP ratios.
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AIM: The aim of this study was to evaluate the effects of various combinations of tropical grass-legume species in rations on the biohydrogenation (BH) activity of unsaturated fatty acids (FAs), C18:0 composition, and fermentation profile in an in vitro rumen system. MATERIALS AND METHODS: Samples of the following five fodder plants were used: One species of grass (Pennisetum purpureum) and four species of tree legumes (Leucaena leucocephala, Gliricidia sepium, Calliandra calothyrsus, and Indigofera zollingeriana). The following eight experimental diets were evaluated: 50% P. purpureum + 50% L. leucocephala (LL I); 50% P. purpureum + 50% G. sepium (GS I); 50% P. purpureum + 50% C. calothyrsus (CC I); 50% P. purpureum + 50% I. zollingeriana (IZ I); 75% P. purpureum + 25% L. leucocephala (LL II); 75% P. purpureum + 25% G. sepium (GS II); 75% P. purpureum + 25% C. calothyrsus (CC II); and 75% P. purpureum + 25% I. zollingeriana (IZ II). Each ration was replicated 3 times. In vitro rumen incubation was performed for 48 h, according to the Tilley and Terry method. Determination of the FA profiles of the forage materials and rumen fluid samples was performed using gas chromatography. RESULTS: The percentage of polyunsaturated FA (PUFA) in the forage materials ranged from 34.18% (P. purpureum) to 74.51% (C. calothyrsus). The percentage of monounsaturated FA (MUFA) ranged from 5.06% (P. purpureum) to 8.71% (L. leucocephala). The percentage of saturated FA (SFA) was the lowest at 19.12% (C. calothyrsus) and highest at 60.76% (P. purpureum). In vitro BH of C18:3 n-3, C18:2 n-6, C18:1 n-9, and C18 PUFA in the experimental diets ranged from 72% to 100%. The BH of C18:1 n-9 in GS I (80%) and IZ I (72%) was significantly different (p<0.05). The percentage of C18:0 was 10-50% and significantly different (p<0.05) among treatments, with the highest (of 50%) in GS II. No significant differences (p>0.05) were observed in the fermentation parameters (pH, total volatile FAs, in vitro dry matter digestibility, and in vitro organic matter digestibility) among the treatments, except in NH3 concentration (p<0.05). CONCLUSION: The various combinations of tropical legumes do not have significant inhibitory effects on the BH of C18:2 n-6, C18:3 n-3, and C18 PUFA after in vitro incubation for 48h. Furthermore, an increase in the tropical legume ratio in the ration tends to suppress C18:0 formation after the fermentation simulation process. IZ I has the potential to reduce C18:1 n-9 (MUFA) disappearance and yield an ideal rumen fermentation profile.