Analisis Potensi Fermentasi Tepung Daun Binahong Anredera Cordifolia Dengan Metode Trial End Error Sebagai Pakan Tambahan Pada Ayam Petelur

Ayam petelur merupakan ternak yang mudah stress dan rentan terhadap penyakit. Guna mengatasi keadaan tersebut perlu dicari alternatif bahan herbal yang tidak menimbulkan residu pada telur ayam yaitu dengan memberikan pakan tambahan fermentasi tepung daun binahong. Binahong merupakan salah satu tanaman yang dapat meningkatkan vitalitas dan daya tahan tubuh. Kandungan fitobiotik pada tepung daun binahong (fenol 85,30 mg/kg, total flafonoid 47,40 mg/kg, saponin 66,00 mg/kg, dan alkaloid 2,60 mg/kg). Unggas yang memiliki sistem pencernaan tunggal kurang baik dalam mencerna pakan berserat kasar tinggi. Kebutuhan serat kasar pada ayam petelur maksimal 7% maka, dilakukan analisis manfaat fermentasi menggunakan EM4. Dalam penelitian terdahulu menyebutkan bahwa EM4 menghasilkan sejumlah besar enzim mencerna serat kasar seperti selulase dan mannase. Keuntungan Lactobacillus dalam EM4 dalam mencerna serat kasar adalah karena bakteri tidak menghasilkan serat kasar dalam aktivitasnya, sehingga mereka lebih efektif dalam menurunkan serat kasar. Fermentasi dapat meningkatkan nilai gizi dan juga palatabilitas. Pemberian pakan tambahan fermentasi tepung daun binahong sebagai feed adiktif memiliki potensi terhadap peningkatkan sistem imun sehingga ayam sehat, maka produksi telur akan meningkat dan stabil. Laying hens are animals that are easily stressed and susceptible to disease. In order to overcome this situation, it is necessary to find alternative herbal ingredients that do not cause residues in chicken eggs, namely by providing additional feed with fermented binahong leaf flour. Binahong is one of the plants that can increase vitality and endurance. Phytobiotic content in binahong leaf flour (phenol 85.30 mg/kg, total flavonoids 47.40 mg/kg, saponins 66.00 mg/kg, and alkaloids 2.60 mg/kg). Poultry that has a single digestive system is not good at digesting high crude fiber feed. The need for crude fiber in laying hens is a maximum of 7%, so an analysis of the benefits of fermentation using EM4 was carried out. In previous studies, it was stated that EM4 produces a large number of enzymes digesting crude fiber such as cellulase and mannase. The advantage of Lactobacillus in EM4 in digesting crude fiber is that bacteria do not produce crude fiber in their activity, so they are more effective in reducing crude fiber. Fermentation can increase nutritional value and also palatability. The addition of fermented binahong leaf flour feed as an addictive feed has the potential to increase the immune system so that chickens are healthy, so egg production will increase and be stable.

Coculture with hemicellulose-fermenting microbes reverses inhibition of corn fiber solubilization by Clostridium thermocellum at elevated solids loadings

Background The cellulolytic thermophile Clostridium thermocellum is an important biocatalyst due to its ability to solubilize lignocellulosic feedstocks without the need for pretreatment or exogenous enzyme addition. At low concentrations of substrate, C. thermocellum can solubilize corn fiber > 95% in 5 days, but solubilization declines markedly at substrate concentrations higher than 20 g/L. This differs for model cellulose like Avicel, on which the maximum solubilization rate increases in proportion to substrate concentration. The goal of this study was to examine fermentation at increasing corn fiber concentrations and investigate possible reasons for declining performance. Results The rate of growth of C. thermocellum on corn fiber, inferred from CipA scaffoldin levels measured by LC–MS/MS, showed very little increase with increasing solids loading. To test for inhibition, we evaluated the effects of spent broth on growth and cellulase activity. The liquids remaining after corn fiber fermentation were found to be strongly inhibitory to growth on cellobiose, a substrate that does not require cellulose hydrolysis. Additionally, the hydrolytic activity of C. thermocellum cellulase was also reduced to less-than half by adding spent broth. Noting that > 15 g/L hemicellulose oligosaccharides accumulated in the spent broth of a 40 g/L corn fiber fermentation, we tested the effect of various model carbohydrates on growth on cellobiose and Avicel. Some compounds like xylooligosaccharides caused a decline in cellulolytic activity and a reduction in the maximum solubilization rate on Avicel. However, there were no relevant model compounds that could replicate the strong inhibition by spent broth on C. thermocellum growth on cellobiose. Cocultures of C. thermocellum with hemicellulose-consuming partners—Herbinix spp. strain LL1355 and Thermoanaerobacterium thermosaccharolyticum—exhibited lower levels of unfermented hemicellulose hydrolysis products, a doubling of the maximum solubilization rate, and final solubilization increased from 67 to 93%. Conclusions This study documents inhibition of C. thermocellum with increasing corn fiber concentration and demonstrates inhibition of cellulase activity by xylooligosaccharides, but further work is needed to understand why growth on cellobiose was inhibited by corn fiber fermentation broth. Our results support the importance of hemicellulose-utilizing coculture partners to augment C. thermocellum in the fermentation of lignocellulosic feedstocks at high solids loading.

Lignocelluloytic activities and composition of bacterial community in the camel rumen

<abstract> <p>The camel is well-adapted to utilize the poor-quality forages in the harsh desert conditions as the camel rumen sustains fibrolytic microorganisms, mainly bacteria that are capable of breaking down the lignocellulosic biomass efficiently. Exploring the composition of the bacterial community in the rumen of the camel and quantifying their cellulolytic and xylanolytic activities could lead to understanding and improving fiber fermentation and discovering novel sources of cellulases and xylanases. In this study, Illumina MiSeq sequencing of the V4 region on 16S rRNA was applied to identify the bacterial and archaeal communities in the rumen of three camels fed wheat straw and broom corn. Furthermore, rumen samples were inoculated into bacterial media enriched with xylan and different cellulose sources, including filter paper (FP), wheat straw (WS), and alfalfa hay (AH) to assess the ability of rumen bacteria to produce endo-cellulase and endo-xylanase at different fermentation intervals. The results revealed that the phylum Bacteroidetes dominated the bacterial community and <italic>Candidatus Methanomethylophilus</italic> dominated the archaeal community. Also, most of the bacterial community has fibrolytic potential and the dominant bacterial genera were <italic>Prevotella</italic>, <italic>RC9_gut_group</italic>, <italic>Butyrivibrio</italic>, <italic>Ruminococcus</italic>, <italic>Fibrobacteres</italic>, and <italic>Treponema</italic>. The highest xylanase production (884.8 mU/mL) was observed at 7 days. The highest cellulase production (1049.5 mU/mL) was observed when rumen samples were incubated with Alfalfa hay for 7 days.</p> </abstract>

Coculture with hemicellulose-fermenting microbes reverses inhibition of corn fiber solubilization by Clostridium thermocellum at elevated solids loadings

BackgroundThe cellulolytic thermophile Clostridium thermocellum is an important biocatalyst due to its ability to solubilize lignocellulosic feedstocks without the need for pretreatment or exogenous enzyme addition. At low concentrations of substrate, C. thermocellum can solubilize corn fiber >95% in 5 days, but solubilization declines markedly at substrate concentrations higher than 20 g/L. This differs for model cellulose like Avicel, on which the maximum solubilization rate increases in proportion to substrate concentration. The goal of this study was to examine fermentation at increasing corn fiber concentrations and investigate possible reasons for declining performance.ResultsThe rate of growth of C. thermocellum on corn fiber, inferred from CipA scaffoldin levels measured by LC-MS/MS, showed very little increase with increasing solids loading. To test for inhibition, we evaluated the effects of spent broth on growth and cellulase activity. The liquids remaining after corn fiber fermentation were found to be strongly inhibitory to growth on cellobiose, a substrate that does not require cellulose hydrolysis. Additionally, the hydrolytic activity of C. thermocellum cellulase was also reduced to less than half by adding spent broth. Noting that >15 g/L hemicellulose oligosaccharides accumulated in the spent broth of a 40 g/L corn fiber fermentation, we tested the effect of various model carbohydrates on growth on cellobiose and Avicel. Some compounds like xylooligosaccharides caused a decline in cellulolytic activity and a reduction in the maximum solubilization rate on Avicel. However, there were no relevant model compounds that could replicate the strong inhibition by spent broth on C. thermocellum growth on cellobiose. Cocultures of C. thermocellum with hemicellulose consuming partners - Herbinix spp. strain LL1355 and Thermoanaerobacterium thermosaccharolyticum –exhibited lower levels of unfermented hemicellulose hydrolysis products, a doubling of the maximum solubilization rate, and final solubilization increased from 67% to 93%. ConclusionsThis study documents inhibition of C. thermocellum with increasing corn fiber concentration and demonstrates inhibition of cellulase activity by xylooligosaccharides, but further work is needed to understand why growth on cellobiose was inhibited by corn fiber fermentation broth. Our results support the importance of hemicellulose-utilizing coculture partners to augment C. thermocellum in the fermentation of lignocellulosic feedstocks at high solids loading.

Pretreatment of Rapeseed Meal Increases Its Recalcitrant Fiber Fermentation and Alters the Microbial Community in an in vitro Model of Swine Large Intestine

The aim of current study was to investigate in an in vitro study how enzymatic and chemical pretreated rapeseed meal (RSM) influences the fiber fermentation and microbial community in the swine large intestine. RSM was processed enzymatically by a cellulase (CELL), two pectinases (PECT), or chemically by an alkaline (ALK) treatment. 16S rRNA gene sequencing data was performed to evaluate changes in the gut microbiota composition, whereas short-chain fatty acid (SCFA) production (ion-chromatography) and non-starch polysaccharides (NSP) composition (using monoclonal antibodies; mAbs) were used to assess fiber degradation. The results showed that ALK, CELL, PECT1, and PECT2 changed microbial community composition, increased the predicted abundance of microbial fiber-degrading enzymes and pathways, and increased acetic acid, propionic acid, butyric acid, and total SCFA production. The increased microbial genera positively correlated with SCFA production. Monoclonal antibody analyses showed that the cell wall polysaccharide structures of RSM shifted after ALK, CELL, PECT1, and PECT2 treatment. The degradation of NSP during the fermentation period was dynamic, and not continuous based on the epitope recognition by mAbs. This study provides the first detailed analysis of changes in the swine intestinal microbiota due to RSM modified by ALK, CELL, PECT1, and PECT2, which altered the microbial community structure, shifted the predicted functional metagenomic profile and subsequently increased total SCFA production. Our findings that ALK, CELL, PECT1, and PECT2 increased fiber degradability in RSM could help guide feed additive strategies to improve efficiency and productivity in swine industry. The current study gave insight into how enzymatic treatment of feed can alter microbial communities, which provides good opportunity to develop novel carbohydrase treatments, particularly in swine feed.

Coculture with Hemicellulose-Fermenting Microbes Reverses Inhibition of Corn Fiber Solubilization by C. Thermocellum at Elevated Solids Loadings

Background The cellulolytic thermophile Clostridium thermocellum is an important biocatalyst due to its ability to solubilize lignocellulosic feedstocks without the need for pretreatment or exogenous enzyme addition. At low concentrations of substrate, C. thermocellum can solubilize corn fiber >95% in 5 days, but solubilization declines markedly at substrate concentrations higher than 20 g/L. This differs for model cellulose like Avicel, on which the maximum solubilization rate increases in proportion to substrate concentration. The goal of this study was to examine fermentation at increasing corn fiber concentrations and investigate possible reasons for declining performance. Results The rate of growth of C. thermocellum on corn fiber, inferred from CipA scaffoldin levels measured by LC-MS/MS, showed very little increase with increasing solids loading. To test for inhibition, we evaluated the effects of spent broth on growth and cellulase activity. The liquids remaining after corn fiber fermentation were found to be strongly inhibitory to growth on cellobiose, a substrate that does not require cellulose hydrolysis. Additionally, the hydrolytic activity of C. thermocellum cellulase was also reduced to less than half by adding spent broth. Noting that >15 g/L hemicellulose oligosaccharides accumulated in the spent broth of a 40 g/L corn fiber fermentation, we tested the effect of various model carbohydrates on growth on cellobiose and Avicel. Some compounds like xylooligosaccharides caused a decline in cellulolytic activity and a reduction in the maximum solubilization rate on Avicel. However, there were no relevant model compounds that could replicate the strong inhibition by spent broth on C. thermocellum growth on cellobiose. Cocultures of C. thermocellum with hemicellulose consuming partners - Herbinix spp. strain LL1355 and Thermoanaerobacterium thermosaccharolyticum –exhibited lower levels of unfermented hemicellulose hydrolysis products, a doubling of the maximum solubilization rate, and final solubilization increased from 67% to 93%. Conclusions This study documents inhibition of C. thermocellum with increasing corn fiber concentration and demonstrates inhibition of cellulase activity by xylooligosaccharides, but further work is needed to understand why growth on cellobiose was inhibited by corn fiber fermentation broth. Our results support the importance of hemicellulose-utilizing coculture partners to augment C. thermocellum in the fermentation of lignocellulosic feedstocks at high solids loading.

Comparative Analysis of Fecal Microbiota in Grasscutter (Thryonomys swinderianus) and Other Herbivorous Livestock in Ghana

The grasscutter (also known as the greater cane rat; Thryonomys swinderianus) is a large rodent native to West Africa that is currently under domestication process for meat production. However, little is known about the physiology of this species. In the present study, aiming to provide information about gut microbiota of the grasscutter and better understand its physiology, we investigated the intestinal microbiota of grasscutters and compared it with that of other livestock (cattle, goat, rabbit, and sheep) using 16S rRNA metagenomics analysis. Similar to the other herbivorous animals, bacteria classified as Bacteroidales, Clostridiales, Ruminococcaceae, and Lachnospiraceae were abundant in the microbiome of grasscutters. However, Prevotella and Treponema bacteria, which have fiber fermentation ability, were especially abundant in grasscutters, where the relative abundance of these genera was higher than that in the other animals. The presence of these genera might confer grasscutters the ability to easily breakdown dietary fibers. Diets for grasscutters should be made from ingredients not consumed by humans to avoid competition for resources and the ability to digest fibers may allow the use of fiber-rich feed materials not used by humans. Our findings serve as reference and support future studies on changes in the gut microbiota of the grasscutter as domestication progresses in order to establish appropriate feeding methods and captivity conditions.

Relationship between Dietary Fiber Fermentation and Volatile Fatty Acids’ Concentration in Growing Pigs

This study was conducted to determine whether differences in fiber fermentation in fiber-rich feed ingredients exist and to assess relationship between fiber fermentation and concentration of volatile fatty acids (VFA) in pig. Castrated males (barrows) were allotted randomly to six diets formulated with different amounts of wheat bran (WB), corn bran (CB), sugar beet pulp (SBP), oat bran (OB), soybean hulls (SH) or rice bran (RB). The apparent ileal digestibility (AID) of soluble dietary fiber (SDF) for OB and SH diets was greater (P < 0.05) than for the other diets. The fermentation of total dietary fiber (TDF) and insoluble dietary fiber (IDF) in the hindgut were greater (P < 0.05) for SBP and SH diets than for WB, CB, OB and RB diets. The apparent total tract digestibility (ATTD) values of all fiber components in SBP, SH and OB diets were greater (P < 0.05) than for WB, CB and RB diets. The concentration of VFA in feces was positively correlated with the ATTD of IDF and cellulose, and ATTD of IDF is the best factor for predicting fecal VFA concentration. Overall, dietary fiber source affected fermentable characteristics of fiber components in the different digestive segments of pig intestine.

Metabolic breath signature of 13C-enriched wheat bran consumption related to gut fermentation in humans: a Fiber-TAG study

IntroductionDietary fibers (DF) have been classified mainly according to their physico-chemical and fermentability properties but it remains unclear whether such classification is relevant when addressing their health effects. Indeed, the nature of physiological effects induced by DF, particularly through their interaction with gut microbiota, remains poorly known due to their diversity, to gut microbiota inter-subjects variability and to the lack of validated non-invasive biomarkers to characterize DF-gut microbiota interaction. The aim of this pilot study was 1) to follow the metabolic fate of 13C-labeled DF through the assessment of 13C-labelled gut-derived metabolites in excreted breath and 2) to evaluate novel non-invasive breath-derived biomarkers of DF-gut microbiota interactions.Materials and methodsSix healthy women (29.7 ± 1.7 years old, BMI: 23.2 ± 0.9kg/m2, fiber intake: 23 ± 1g/d) consumed in research settings a controlled breakfast containing eight 13C-labelled wheat bran biscuits (50 g of labelled wheat bran, 3.0At%13C). 13C-labelled wheat bran was obtained from wheat cultivated under 13CO2 enriched atmosphere. Samples of expired gases were collected during 24 h after ingestion in order to measure H2 and CH4 by gas chromatography (GC) with piezoelectric detection and 13CO2 and 13CH4 by gas chromatography coupled with an isotope ratio mass spectrometer (GC-IRMS). Apart test breakfast, subjects only consumed standardized meals without fibers.ResultsThe analysis of H2 and CH4 24h-kinetic measurements distinguished 2 groups in terms of fermentation related gas excretion: the high-CH4 producers with high baseline CH4 concentrations (42.1 ± 13.7ppm) and low baseline H2 concentrations (7.3 ± 5.8ppm) and the low-CH4 producers with low baseline CH4 concentrations (6.5 ± 3.6ppm) and high baseline H2 concentrations (20.8 ± 16.0ppm). Following the 13C-wheat bran biscuits’ ingestion, postprandial H2 and CH4 concentrations increased more significantly in the high-CH4 producer subjects. 13C enrichment was detectable in expired gases in all subjects. 13CO2 kinetics were similar for all subjects and correspond to the oxidation of the digestible part of the bran. The appearance of 13CH4 was significantly enhanced and prolonged after 180 min in high-CH4 producers compared to low-CH4 producers, suggesting distinct fiber fermentation profile.DiscussionThis pilot study allowed to consider novel procedures for development of non-invasive breath biomarkers of fiber-gut microbiota interactions. Assessment of expired gas excretion following 13C-labelled fiber ingestion allowed deciphering distinct fermentation profiles: high-CH4 producers vs low-CH4 producers and accordingly provide a related non-invasive breath metabolic signature of the fiber fermentation for each profile. Further gut microbiota and 13C-metabolites analysis will permit to relate the gut bacteria composition with breath gas excretion kinetics according to fiber fermentation profile.