In Vitro Fermentation, 2nd Edition

A special issue of Fermentation (ISSN 2311-5637). This special issue belongs to the section "Microbial Metabolism, Physiology & Genetics".

Deadline for manuscript submissions: closed (31 March 2023) | Viewed by 32110

Special Issue Editor


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Guest Editor
College of Animal Science and Technology, Yangzhou University, Yangzhou, China
Interests: ruminant; rumen; caecum; microbiology; bacteria; protozoa; fermentation; fermenter; VFA; amino acid
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

In vitro fermentation is a critical technology for studying and evaluating feed nutritional value, nutrient metabolism mechanism and fermentation state. In vitro fermentation includes simulated rumen in vitro fermentation, in vitro gas production, in vitro microbial fermentation, in vitro enzyme fermentation, combined two-step fermentation, etc. Over the years, in vitro fermentation has evaluated the feeding value of a large number of feeds to ruminants and monogastric animals such as swine. It not only enriches the feed database, but also makes an important contribution to the development and utilization of microbial resources and the improvement of feed utilization rate.

Substrate, microorganism, enzyme and fermentation state are important elements of in vitro fermentation, and equipment and fermentation regulation of in vitro fermentation are also important guarantees for fermentation. Therefore, the goal of this Special Issue is to publish both recent innovative research results and review papers on in vitro fermentation with nutrients or new feed resources. Review and research papers on the development of novel enzymes, microbial strains, and fermentation equipment are also of interest. If you would like to contribute a review paper, please contact one of the editors to discuss the relevance of the topic before submitting the manuscript.

Prof. Dr. Mengzhi Wang
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Fermentation is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2600 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • microbe
  • microbial ecology
  • nutrients
  • digestibility
  • enzyme
  • metabolite
  • gas
  • rumen
  • fermenter
  • medium

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Published Papers (19 papers)

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Research

23 pages, 5208 KiB  
Article
In Vitro Fermentation of Different Indigestible Glucans with Varying Physico-Chemical Properties by Human Fecal Microbiota
by Hao Zhang, Chunhua Chen, Yanli Zhang, Hongmei Yin, Jielun Hu, Yadong Zhong, Shaoping Nie and Mingyong Xie
Fermentation 2023, 9(5), 485; https://doi.org/10.3390/fermentation9050485 - 18 May 2023
Cited by 1 | Viewed by 1571
Abstract
Indigestible glucans (IGs) are dietary fibers that can promote human health via fermentation by the gut microbiota, where their physico-chemical properties play a crucial role. This effect remains to be fully explored. The aim of the current study was to comprehensively investigate and [...] Read more.
Indigestible glucans (IGs) are dietary fibers that can promote human health via fermentation by the gut microbiota, where their physico-chemical properties play a crucial role. This effect remains to be fully explored. The aim of the current study was to comprehensively investigate and compare the fermentation characteristics of IGs with various structural properties, as well as their effects on the gut microbiota. Barley β-glucan (BG), laminarin (L), yeast β-glucan (BY), pachyman (PAC), resistant starch (R), and litesse (Lit) were anaerobically batch-fermented using the human fecal microbiota for 48 h. All the IGs were utilized by the gut microbiota at different rates, and 2% of L, 14% of BG, 23% of BY and PAC, and 35% of R and Lit were unfermented at the 48th hour. During fermentation, mono-, di-, or trisaccharides were released from BG, L, and Lit, and the pH of broth was greatly lowered by IGs, especially BG and L, along with the production of short-chain fatty acids. Interestingly, PAC favored butyric acid production, while BG, L, and BY preferred propionic acid. Moreover, lactic acid, but not succinic acid, was detected in considerable amounts, but only with BG and L after 5 h. 16S rDNA analysis showed different microbial structures and the selective promotion of bacteria with different IGs, while only PAC did not decrease microbial α-diversity. Further qPCR analysis confirmed that BG was more potent at proliferating Faecalibacterium prausnitzii; BY preferred total bacteria, Prevotella, and Lactobacillus; and R favored Bifidobacterium. The IG-induced changes in the gut microbiota were strongly correlated with carboxylic acid production. In conclusion, the six IGs differed in fermentation characteristics and gut microbiota regulation capacity, and each one could have specific applications in human health promotion. Full article
(This article belongs to the Special Issue In Vitro Fermentation, 2nd Edition)
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16 pages, 2950 KiB  
Article
Calm Hu Sheep Have a Different Microbiome Profile and Higher Energy Utilization Efficiency Than Nervous Hu Sheep
by Feifan Wu, Luoyang Ding, Jiasheng Wang, Qiaoqing Chen, Asmita Thapa, Jianyu Mao and Mengzhi Wang
Fermentation 2023, 9(5), 470; https://doi.org/10.3390/fermentation9050470 - 13 May 2023
Viewed by 1363
Abstract
In sheep, temperament is known to affect animal welfare and the quality of animal products. While the composition of the gut microbiota is different between depressed patients and healthy human patients, in sheep, the influence of temperament on ruminal microbial species and abundance [...] Read more.
In sheep, temperament is known to affect animal welfare and the quality of animal products. While the composition of the gut microbiota is different between depressed patients and healthy human patients, in sheep, the influence of temperament on ruminal microbial species and abundance remains unknown. This study investigated the effects of temperament on parameters of rumen fermentation and microbial composition of rumen contents of Hu ram lambs. Using the pen score test, 6 lambs that scored 2 points or below (calm) and 6 lambs that scored 4 points or more (nervous) were selected from 100 ram lambs. The sheep were fed a standard diet for 60 days and rumen samples were collected at slaughter. The concentrations of propionic acid, isovaleric acid, valeric acid, and the ammonia nitrogen concentration were different between the calm and the nervous groups (p < 0.05). At the phylum level, there were significant differences in Bacteroidetes, Tenericutes, and Spirochetes (p < 0.05); and at the genus level, there were significant differences in the Christensenellaceae R-7 group, Treponema 2, Fibrobacter, and Ruminococcaceae UCG-003 (p < 0.05). The present study suggests that differences in the rumen microbiota between the calm group and the nervous group could have an impact on the metabolism of carbohydrates and polysaccharides and explain why Calm Hu sheep have a higher energy utilization efficiency than nervous Hu sheep. More studies are needed to further understand the effect of temperament on specific pathways of the rumen microbiota. Full article
(This article belongs to the Special Issue In Vitro Fermentation, 2nd Edition)
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11 pages, 324 KiB  
Article
In Vivo Digestibility and In Vitro Fermentation of High Dietary Fiber Forages in Growing Pigs’ Diets
by Mónica Gandarillas, María Isidora Valenzuela, Jorge Molina, Rodrigo Arias and Juan Keim
Fermentation 2023, 9(5), 448; https://doi.org/10.3390/fermentation9050448 - 09 May 2023
Viewed by 1034
Abstract
The pig farming industry is constantly challenged to seek low-cost ingredients that fulfill animal requirements. In this study, two summer forage brassica meals were assessed as sources of dietary fiber in growing pigs by in vivo digestibility and in vitro fermentation experiments. The [...] Read more.
The pig farming industry is constantly challenged to seek low-cost ingredients that fulfill animal requirements. In this study, two summer forage brassica meals were assessed as sources of dietary fiber in growing pigs by in vivo digestibility and in vitro fermentation experiments. The control diet included corn, soybean meal, and wheat middlings. The experimental diets replaced wheat middlings (15%) with turnip (Brassica rapa) roots or fodder rape (Brassica napus) whole plant meal, respectively. All diets were elaborated to be iso-nitrogenous and iso-energetic. The turnip diet had a greater digestibility rate for gross energy (p = 0.020). The ash digestibility was greater for the rape diet and intermediate for the turnip diet, with the lowest value for the control diet (p = 0.003). When incubating pure brassica forages, only gas production at 72 h was greater for the turnip than rape diet (p = 0.04). No differences (p > 0.05) in the in vitro gas production parameters were observed among the diets. The pure fermentation of turnip increased the VFA concentration and propionate molar proportion, whereas acetate was reduced (p < 0.05), which resulted in a trend towards a greater propionate molar proportion with the inclusion of turnip in the diet (p = 0.067). The inclusion of 15% of turnip meal increased the in vivo energy digestibility and tended to modify the fermentation parameters, increasing the molar proportion of propionate, whereas the inclusion of whole plant fodder rape did not affect the in vivo digestibility or in vitro fermentation compared with the control diet. Full article
(This article belongs to the Special Issue In Vitro Fermentation, 2nd Edition)
14 pages, 1804 KiB  
Article
Degradability of Vicia ervilia Grain Using In Situ and CNCPS Methods, and Model-Based Analysis of Its Ruminal Degradation
by Marziyeh Taghavi, Akbar Taghizadeh, Yousef Mehmannavaz, Ali Hoseinkhani, Hamid Mohammadzadeh, Muhlis Macit, Valiollah Palangi and Maximilian Lackner
Fermentation 2023, 9(5), 419; https://doi.org/10.3390/fermentation9050419 - 27 Apr 2023
Viewed by 1166
Abstract
Using nylon bag techniques, Cornell net carbohydrates and protein systems (CNCPS), and scanning electron microscopy, the authors examined the digestibility and structure of Vicia ervilia (ervil, bitter vetch) after steam flaking, roasting, and microwave processing. During the in situ technique, the samples were [...] Read more.
Using nylon bag techniques, Cornell net carbohydrates and protein systems (CNCPS), and scanning electron microscopy, the authors examined the digestibility and structure of Vicia ervilia (ervil, bitter vetch) after steam flaking, roasting, and microwave processing. During the in situ technique, the samples were incubated at 0, 2, 4, 6, 8, 12, 16, 24, 36, and 48 h. For the description of the ruminal DM (dry matter) and CP (crude protein) degradation kinetics of treated and untreated Vicia ervilia, different models were selected as the best fit for the dry matter (DM) and crude protein (CP) degradation parameters of steam flaked samples. The results showed that both the steam flaking and microwave treatment samples contained high levels of non-protein nitrogen and buffer soluble protein, respectively. In comparison with steam flaking and microwave treatment, roasting decreased and increased the buffer soluble protein and neutral detergent insoluble protein, respectively. The control treatments showed the highest levels of neutral detergent soluble protein and the lowest levels of acid detergent soluble protein. Moreover, steam flaking and roasting decreased and increased the amount of acid detergent insoluble protein, respectively. When using dry heat (microwave and roasting), the acid detergent soluble and insoluble protein fractions were increased. As a result of this experiment, the nitrogen fractions were altered using heat processing. Hence, protein fermented feed and ruminal fermentation conditions can be expressed using these results. Full article
(This article belongs to the Special Issue In Vitro Fermentation, 2nd Edition)
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15 pages, 912 KiB  
Article
In Vitro Rumen Fermentation Kinetics Determination and Nutritional Evaluation of Several Non-Conventional Plants with Potential for Ruminant Feeding
by Cristiana S. A. M. Maduro Dias, Helder Nunes, Carlos Vouzela, João Madruga and Alfredo Borba
Fermentation 2023, 9(5), 416; https://doi.org/10.3390/fermentation9050416 - 26 Apr 2023
Cited by 2 | Viewed by 1457
Abstract
Using invasive plants as non-conventional forage in livestock production can contribute to meeting climate change targets and provide a competitive advantage in a global market. This study evaluated the nutritional potential of several non-conventional plants: Pennisetum setaceum, Ricinus communis, Arundo donax, Acacia [...] Read more.
Using invasive plants as non-conventional forage in livestock production can contribute to meeting climate change targets and provide a competitive advantage in a global market. This study evaluated the nutritional potential of several non-conventional plants: Pennisetum setaceum, Ricinus communis, Arundo donax, Acacia melanoxylon, Opuntia ficus-indica, Agave americana, Pittosporum undulatum, and Hedychium gardnerianum. Chemical composition, in vitro digestibility, gas production, and energy estimates were determined. Opuntia showed the lowest DM value (6.65%), while the highest was found for Acacia (39.43%). Ricinus recorded the highest levels of CP (23.56% DM), RFV (273.86), and in vitro gas production at 24 h (43.49 mL/200 mg DM). The highest NDF (80.39% DM), HEM (39.03% DM), and CEL (36.81% DM) values were observed for Pennisetum. Agave produced the highest amount of gas from the start to the end of incubation (22.68 to 48.99 mL/200 mg DM), while Acacia produced the least (3.83 to 14.78 mL/200 mg DM). The highest ME (8.72 MJ/kg DM) and NEL (5.06 MJ/kg DM) estimates were obtained for Agave. Correlations between the chemical compositions and feed quality indices of the plants were observed, showing strong negative correlations between ADF and DMD, OMD, and DMI (r > 0.86, p < 0.01). We can conclude that all these non-conventional plants have potential as an alternative feed for ruminants when there are fodder shortages. Full article
(This article belongs to the Special Issue In Vitro Fermentation, 2nd Edition)
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13 pages, 312 KiB  
Article
Evaluation of Fermentability of Whole Soybeans and Soybean Oligosaccharides by a Canine In Vitro Fermentation Model
by Hee Seong Kim, Evan C. Titgemeyer and Charles Gregory Aldrich
Fermentation 2023, 9(5), 414; https://doi.org/10.3390/fermentation9050414 - 26 Apr 2023
Cited by 3 | Viewed by 1412
Abstract
Soybean oligosaccharides (OS) have been recognized as a prebiotic that can be fermented in the colon, resulting in short-chain fatty acid (SCFA) production that can be used as an energy source for colonocytes, supporting cell differentiation and gut health. The objective was to [...] Read more.
Soybean oligosaccharides (OS) have been recognized as a prebiotic that can be fermented in the colon, resulting in short-chain fatty acid (SCFA) production that can be used as an energy source for colonocytes, supporting cell differentiation and gut health. The objective was to determine the effects of WSBOS on in vitro fermentation, using dog feces as inoculum. Treatments included total dietary fiber (TDF) residues from WSB, soybean hulls (SH), pea fiber (PF), and beet pulp (BP), as well as WSB TDF residue plus soybean OS (WSBOS) and WSB TDF residue plus raffinose, stachyose, and verbascose (WSBRSV). Fresh fecal samples were collected from dogs and maintained in anaerobic conditions until substrate inoculation. Test tubes containing fiber sources and inoculum were incubated for 4, 8, and 12 h at 39 °C. Organic matter disappearance (OMD), pH, and SCFA were measured. The WSBOS and WSBRSV had greater (p < 0.05) OMDs than BP. Butyrate production was greatest (p < 0.05) for WSBOS (294.7 µmol/g) and WSBRSV (266.1 µmol/g), followed by BP (130.3 µmol/g) and WSB (109.2 µmol/g), and lowest (p < 0.05) for PF (44.1 µmol/g). The production of total SCFA was greatest (p < 0.05) for BP and WSBOS, followed by WSB, and lowest (p < 0.05) for PF. In conclusion, WSB has the potential as a prebiotic demonstrating greater butyrate production than BP in a canine in vitro fermentation model due to the fermentation of both OS and fiber in WSB. Further animal feeding studies are needed to determine the appropriate amount of WSB in canine diets. Full article
(This article belongs to the Special Issue In Vitro Fermentation, 2nd Edition)
12 pages, 1094 KiB  
Article
Saccharomyces cerevisiae Culture’s Dose–Response Effects on Ruminal Nutrient Digestibility and Microbial Community: An In Vitro Study
by Dongwen Dai, Yanfang Liu, Fanlin Kong, Cheng Guo, Chunxiao Dong, Xiaofeng Xu, Shengli Li and Wei Wang
Fermentation 2023, 9(5), 411; https://doi.org/10.3390/fermentation9050411 - 26 Apr 2023
Viewed by 1394
Abstract
Supplementation with saccharomyces cerevisiae culture products (SCs) has shown effectiveness in alleviating or improving the health and productivity of ruminants at a high risk of digestive and metabolic problems as a consequence of their physiological state and feeding system (i.e., Holstein cows during [...] Read more.
Supplementation with saccharomyces cerevisiae culture products (SCs) has shown effectiveness in alleviating or improving the health and productivity of ruminants at a high risk of digestive and metabolic problems as a consequence of their physiological state and feeding system (i.e., Holstein cows during peak lactation). However, the effects of SC supplementation on ruminal digestion and microbial population are not yet well-understood. Hence, this study aimed to contribute to the knowledge of the effects of in vitro SC supplementation on ruminal nutrient digestibility and microbial community. This study included three treatment groups: a control group (CON, 0% SC proportion of substrate DM), a low-dose SC group (LSC, 0.10% SC proportion of substrate DM), and a high-dose SC group (HSC, 0.30% SC proportion of substrate DM). The SC product contained 7.0 × 109 CFU/g. After 48 h of fermentation at 39 °C, the incubation fluid and residue were collected to measure the ruminal nutrient digestibility and microbial community. The results showed that supplemental SC tended (p = 0.096) to increase DM digestibility due to an increase (9.6%, p = 0.03) in CP digestibility and via a tendency (0.05 < p < 0.08) to increase the fiber fraction. Additionally, the 16S rRNA high-throughput sequencing results revealed that the richness and diversity of the microbiota were unchanged by SC supplementation, while the abundances of Spirochaetes, Tenericutes, and Spirochaetaceae were lower in the SC groups than those in the CON group (p < 0.05). At the genus level, the abundances of Selenomonas and Succinivibrio were increased by SC supplementation (p < 0.05), while SC supplementation decreased the abundances of Ruminococcaceae_UCG-014 and Treponema_2 (p < 0.05). Furthermore, the predicted function of the microbiota showed that carbohydrate metabolism and lipid metabolism were enriched in the SC groups compared with the CON group (p < 0.05). Except for the increases in ADF digestibility (p = 0.032) and pH (p = 0.076) at 0.30%, the supplemental level did not result in additional effects. In summary, our results demonstrate that SC supplementation could improve ruminal nutrient degradation digestibility and alter microbiota composition. Full article
(This article belongs to the Special Issue In Vitro Fermentation, 2nd Edition)
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12 pages, 1996 KiB  
Article
Impact of Rumen Fluid Storage on In Vitro Feed Fermentation Characteristics
by Bereket Zeleke Tunkala, Kristy DiGiacomo, Pablo S. Alvarez Hess, Frank R. Dunshea and Brian J. Leury
Fermentation 2023, 9(4), 392; https://doi.org/10.3390/fermentation9040392 - 19 Apr 2023
Cited by 4 | Viewed by 1385
Abstract
Storing rumen fluid (RF) has the potential to standardize subsequent in vitro feed fermentation studies. The first phase of this experiment aimed to evaluate the effect of two RF storage methods on gas composition and dry matter disappearance (DMD) in wheat grain and [...] Read more.
Storing rumen fluid (RF) has the potential to standardize subsequent in vitro feed fermentation studies. The first phase of this experiment aimed to evaluate the effect of two RF storage methods on gas composition and dry matter disappearance (DMD) in wheat grain and lucerne hay under in vitro fermentation. The storage methods were as follows: (1) snap-freezing RF using liquid nitrogen and then storing it at −80 °C (−80 °C); and (2) mixing RF with 5% dimethyl sulfoxide (DMSO), subsequently freezing it at −20 °C (D−20 °C), and comparing it to fresh RF on days 1, 14, and 30 post collection. The objective of the second phase was to quantify the impact of preserving the RF for 180 days at D−20 °C on the in vitro fermentation parameters. The methane composition was lower (p < 0.001) in both the preserved RFs than in the fresh RF. There was no difference (p < 0.05) in DMD values between days 14 and 30. The average cumulative gas production and DMD from the RF stored at D−20 °C was higher than that from the RF stored at −80 °C. Moreover, there was no difference between day 30 and day 180 in the total gas production and lag time when fermenting with RF preserved at D−20 °C. Therefore, storing RF at D−20 °C is preferable to storing it at −80 °C when access to fresh RF is limited. Full article
(This article belongs to the Special Issue In Vitro Fermentation, 2nd Edition)
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19 pages, 5222 KiB  
Article
Lactobacillus acidophilus Fermented Dandelion Improves Hyperuricemia and Regulates Gut Microbiota
by Qianwen Ma, Mingju Chen, Yu Liu, Ying Tong, Tianfeng Liu, Lele Wu, Jiliang Wang, Bin Han, Lin Zhou and Xuguang Hu
Fermentation 2023, 9(4), 352; https://doi.org/10.3390/fermentation9040352 - 03 Apr 2023
Cited by 2 | Viewed by 2428
Abstract
Foodborne prevention and treatment of hyperuricemia (HUA) has received widespread attention. Lactic acid bacteria (LAB) can improve intestinal function, while traditional medicine dandelion has the functions of detoxification and detumescence. Whether LAB fermented dandelion has any effects on HUA and the underlying mechanism [...] Read more.
Foodborne prevention and treatment of hyperuricemia (HUA) has received widespread attention. Lactic acid bacteria (LAB) can improve intestinal function, while traditional medicine dandelion has the functions of detoxification and detumescence. Whether LAB fermented dandelion has any effects on HUA and the underlying mechanism is not clear. To address these questions, Lactobacillus acidophilus was selected or maximal xanthine oxidase activity. The effect of Lactobacillus acidophilus fermented dandelion (LAFD) on uric acid metabolism was evaluated by the HUA mouse model. Expression levels of UA, BUN, CRE, XOD, and inflammatory factors in serum were detected. Paraffin sections and staining were used to observe the kidney and small intestine, and mRNA expression of GLUT9, URAT1, OAT1, and ABCG2 related to uric acid metabolism were investigated. Furthermore, the intestinal flora was studied by contents of the cecum and high throughput 16S rRNA sequencing. The results showed that LAFD had a significant inhibitory effect on XOD in vitro (p < 0.01). LAFD could reduce the levels of UA, BUN, CRE, XOD, IL-1 β, IL-6, and TNF- α in serum (p < 0.05), thus inhibiting inflammatory reaction, and reducing UA by decreasing the mRNA expression of GLUT9, URAT1 in kidney and increasing the mRNA expression of OAT1 and ABCG2 in kidney and small intestine (p < 0.05). In addition, the 16S rRNA gene sequencing analysis demonstrated that LAFD treatment can help restore the imbalance of the intestinal microbial ecosystem and reverse the changes in Bacterodietes/Firmicutes, Muribaculaceae, Lachnospiraceae in mice with HUA. It is suggested that the mechanism of LAFD in treating HUA may be related to the regulation of the mRNA expressions of GLUT9, URAT1, OAT1, and ABCG2 in the kidney and small intestine, as well as the regulation of intestinal flora, which provides the experimental basis for the development of new plant fermented products. Full article
(This article belongs to the Special Issue In Vitro Fermentation, 2nd Edition)
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15 pages, 868 KiB  
Article
The Effect of Direct-Fed Microbials on In-Vitro Rumen Fermentation of Grass or Maize Silage
by Rajan Dhakal, Giuseppe Copani, Bruno Ieda Cappellozza, Nina Milora and Hanne Helene Hansen
Fermentation 2023, 9(4), 347; https://doi.org/10.3390/fermentation9040347 - 01 Apr 2023
Cited by 6 | Viewed by 1915
Abstract
Direct-fed microbial products (DFM) are probiotics that can be used advantageously in ruminant production. The in vitro gas production technique (IVGPT) is a method to simulate rumen fermentation and can be used to measure degradation, gas production, and products of fermentation of such [...] Read more.
Direct-fed microbial products (DFM) are probiotics that can be used advantageously in ruminant production. The in vitro gas production technique (IVGPT) is a method to simulate rumen fermentation and can be used to measure degradation, gas production, and products of fermentation of such additives. However, inter-laboratory differences have been reported. Therefore, tests using the same material were used to validate laboratory reproducibility. The objective of this study was to assess the effect of adding two DFM formulations on fermentation kinetics, methane (CH4) production, and feed degradation in two different basal feeds while validating a newly established IVGPT laboratory. Six treatments, with three replicates each, were tested simultaneously at the established IVGPT lab at the University of Copenhagen, and the new IVGPT lab at Chr. Hansen Laboratories. Maize silage (MS) and grass silage (GS) were fermented with and without the following DFM: P1: Ligilactobacillus animalis and Propionibacterium freudenreichii (total 1.5 × 107 CFU/mL), P2: P1 with added Bacillus subtilis and B. licheniformis (total 5.9 × 107 CFU/mL). The DFM were anaerobically incubated in rumen fluid and buffer with freeze-dried silage samples for 48 h. Total gas production (TGP: mL at Standard Temperature and Pressure/gram of organic matter), pH, organic matter degradability (dOM), CH4concentration (MC) and yield (MY), and volatile fatty acid (VFA) production and profiles were measured after fermentation. No significant differences between the laboratories were detected for any response variables. The dOM of MS (78.3%) was significantly less than GS (81.4%), regardless of the DFM added (P1 and P2). There were no significant differences between the effects of the DFM within the feed type. MS produced significantly more gas than GS after 48 h, but GS with DFM produced significantly more gas at 3 and 9 h and a similar gas volume at 12 h. Both DFM increased TGP significantly in GS at 48 h. There was no difference in total VFA production. However, GS with and without probiotics produced significantly more propionic acid and less butyric acid than MS with and without probiotics. Adding P2 numerically reduced the total methane yield by 4–6% in both MS and GS. The fermentation duration of 48 h, used to determine maximum potential dOM, may give misleading results. This study showed that it is possible to standardize the methodology to achieve reproducibility of IVGPT results. Furthermore, the results suggest that the P2 DFM may have the potential to reduce CH4 production without affecting organic matter degradation. Full article
(This article belongs to the Special Issue In Vitro Fermentation, 2nd Edition)
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19 pages, 1120 KiB  
Article
Differential Diet and pH Effects on Ruminal Microbiota, Fermentation Pattern and Fatty Acid Hydrogenation in RUSITEC Continuous Cultures
by Julio Ernesto Vargas, Lorena López-Ferreras, Sonia Andrés, Iván Mateos, Egon Henrique Horst and Secundino López
Fermentation 2023, 9(4), 320; https://doi.org/10.3390/fermentation9040320 - 23 Mar 2023
Cited by 2 | Viewed by 2300
Abstract
The aim of this study was to distinguish effects due to diet composition from those triggered by ruminal pH on fermentation patterns and microbial profiles in a continuous culture system (RUSITEC). The study followed a 2 × 2 factorial design, with two diets [...] Read more.
The aim of this study was to distinguish effects due to diet composition from those triggered by ruminal pH on fermentation patterns and microbial profiles in a continuous culture system (RUSITEC). The study followed a 2 × 2 factorial design, with two diets varying in the proportions of forage and concentrate and two pH levels in the culture medium. RUSITEC fermenters were used to simulate rumen fermentation and feed digestibility, fermentation end-products, microbial protein synthesis, microbial community, and long-chain fatty acid profiles in the digesta were determined. Multivariate analyses were applied to summarize the overall results. High concentrate (34% cereal grain, 32% hay) diets were more digestible (p < 0.05) than high forage (10% cereal grain, 78% hay) diets, resulting in a greater (p < 0.05) formation of most fermentation end-products and microbial protein in the rumen. However, there were no significant (p > 0.05) differences between diets in methane production. Ciliate protozoa, anaerobic fungi, some fibrolytic bacteria, hydrogenation of oleic acid, and relative proportion of conjugated linoleic acid were increased (p < 0.05) with high forage diets. A decline in rumen pH from 6.8 to 6.4 decreased (p < 0.05) feed digestibility, protein degradability, and the daily outputs of some fermentation end-products (gas, VFA, acetate, ammonia) but had no effect (p > 0.05) on the synthesis of microbial protein, and on the output of methane, propionate, butyrate or lactate. Minor changes in microbial community profile or the fatty acid relative proportions were observed within this pH range. The overall multivariate analysis revealed a clear discrimination between high-concentrate and high-forage diets, with subtler and less-defined pH effects on ruminal fermentation and microbial communities. Full article
(This article belongs to the Special Issue In Vitro Fermentation, 2nd Edition)
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11 pages, 280 KiB  
Article
Greenhouse Gas Emission Reduction Potential of Lavender Meal and Essential Oil for Dairy Cows
by Levend Coşkuntuna, Maximilian Lackner, Kadir Erten, Sevilay Gül, Valiollah Palangi, Fisun Koç and Selim Esen
Fermentation 2023, 9(3), 253; https://doi.org/10.3390/fermentation9030253 - 04 Mar 2023
Cited by 1 | Viewed by 1581
Abstract
This research aims to evaluate the potential of lavender meal (LM) and lavender essential oil (LEO) to mitigate methane emissions by dairy cows. Locally grown lavender was collected fresh for this purpose, and its oil was extracted using the cold-press method. The resultant [...] Read more.
This research aims to evaluate the potential of lavender meal (LM) and lavender essential oil (LEO) to mitigate methane emissions by dairy cows. Locally grown lavender was collected fresh for this purpose, and its oil was extracted using the cold-press method. The resultant LEO and LM and whole lavender (WL) were added to dairy cow concentrate feed at 0%, 0.05%, and 0.10%, and their effects on vitro gas production values and gas concentrations were subsequently assessed. Out of the 30 bioactive compounds isolated from LEO, linalool and linalyl acetate were the most common—accounting for 70.4% of the total. The lavender dose had a significant influence on gas production for up to 12 h. No significant variations were found across the lavender forms when gas kinetics, in vitro degradability, and predicted energy values were compared. The addition of WL to the concentrate feed of dairy cows produced the greatest quantities of methane, carbon dioxide, and hydrogen sulfide, whereas LEO resulted in the lowest values. In contrast, no significant difference in ammonia content was found across the various lavender forms added into dairy cow concentrate feed. The results of this research suggest that adding 0.05–0.10% LM and LEO to concentrate feed may decrease greenhouse gas emissions from dairy cows. Full article
(This article belongs to the Special Issue In Vitro Fermentation, 2nd Edition)
9 pages, 268 KiB  
Article
Effects of Capsicum oleoresin Inclusion on Rumen Fermentation and Lactation Performance in Buffaloes (Bubalus bubalis) during Summer: In Vitro and In Vivo Studies
by Zhigao An, Junwei Zhao, Xinxin Zhang, Shanshan Gao, Chao Chen, Kaifeng Niu, Pei Nie, Zhiqiu Yao, Ke Wei, Umair Riaz and Liguo Yang
Fermentation 2023, 9(3), 232; https://doi.org/10.3390/fermentation9030232 - 28 Feb 2023
Viewed by 1333
Abstract
This research aimed to evaluate the effects of Capsicum oleoresin (CAP) supplementation on rumen fermentation in vivo and In vitro, and lactation performance in buffaloes. In the experiment in vitro, 2 × 5 factorial design was carried out according to two temperatures (normal [...] Read more.
This research aimed to evaluate the effects of Capsicum oleoresin (CAP) supplementation on rumen fermentation in vivo and In vitro, and lactation performance in buffaloes. In the experiment in vitro, 2 × 5 factorial design was carried out according to two temperatures (normal temperature: 39 °C; hyperthermal temperature: 42 °C) and five CAP concentrations (0 mg/L; 2 mg/L; 20 mg/L; 200 mg/L; 2000 mg/L). In the experiment in vivo, four multiparous mid-lactating Mediterranean buffaloes (body weight: 640.08 ± 17.90 kg) were randomly allocated to four treatments according to 4 × 4 Latin square design for CAP supplementation in four dosages (0 mg/kg, 10 mg/kg, 20 mg/kg, or 40 mg/kg of dry matter). The experiment’s results In vitro showed that hyperthermal temperature affected all fermentation characteristics measured in this research. CAP decreased the pH, short-chain fatty acids concentration, and percentages of propionate, butyrate, isobutyrate, valerate, and caproate, while increasing the percentage of acetate and the ratio of acetate to propionate at normal temperature (p ≤ 0.05). In the experiment in vivo, CAP decreased the percentage of propionate and quadratically affected acetate percentage in rumen fluid (p ≤ 0.05). CAP reduced rectal temperature and respiratory rates (p ≤ 0.05) and tended to increase dry matter intake quadratically (p ≤ 0.10). For lactation performance, CAP increased milk yield and milk lactose yield (p ≤ 0.05), and tended to increase milk protein yield (p ≤ 0.10). In conclusion, CAP modified rumen fermentation characteristics in vivo and In vitro and had beneficial effects on lactation performance in buffaloes during summer. Full article
(This article belongs to the Special Issue In Vitro Fermentation, 2nd Edition)
10 pages, 266 KiB  
Article
In Vitro Studies on Rumen Fermentation and Methanogenesis of Different Microalgae and Their Effects on Acidosis in Dairy Cows
by Ekin Sucu
Fermentation 2023, 9(3), 229; https://doi.org/10.3390/fermentation9030229 - 27 Feb 2023
Cited by 3 | Viewed by 1734
Abstract
Two in vitro studies were carried out on nonlactating dairy cows. Experiment 1 compared the methanogenesis and rumen fermentation parameters of various microalgae (Spirulina platensis, Chlorella vulgaris, and Schizochytrium spp.) and protein feeds (sunflower meal, soybean meal, and alfalfa hay) [...] Read more.
Two in vitro studies were carried out on nonlactating dairy cows. Experiment 1 compared the methanogenesis and rumen fermentation parameters of various microalgae (Spirulina platensis, Chlorella vulgaris, and Schizochytrium spp.) and protein feeds (sunflower meal, soybean meal, and alfalfa hay) with monensin (MON). Rumen fermentation parameters were determined by an in vitro gas production system. Experiment 2 compared the ability of three microalgae to prevent acidosis. They were tested for 6 h against oat straw (100 mg) and MON (12 g/mL) to ameliorate ruminal acidosis caused by the addition of glucose (0.1 g/mL) as a fermentable carbohydrate with rumen fluid. In experiment 1, there were variations in the nutrient content of microalgae and protein sources. The dry matter content of the substrates ranged from 90 to 94%, and the organic matter content ranged from 82 to 88%, with Schizochytrium spp. having the highest. Protein content in algae and protein feeds ranged from 18–62% of dry matter (DM) to 16–48% DM, with S. platensis and C. vulgaris having the highest. The ether extract of Schizochytrium spp. (45.5% DM) was the highest of any substrate. In vitro rumen fermentation revealed that protein feeds increased the cumulative gas production at the highest level while MON caused a decrease. Ruminal pH was found to be higher in MON (6.95) and protein feeds (6.77–6.81) than in algae (6.37–6.50). In addition, in terms of metabolizable energy and digestible organic matter, protein feeds outperformed algae. The MON produced the least amount of methane (CH4) of any substrate, but Schizochytrium spp. demonstrated potential for CH4 reduction. In these groups, the decrease in CH4 production was accompanied by a decrease in total volatile fatty acids, acetate, and the acetate-to-propionate ratio, but an increase in propionate. Experiment 2 revealed MON as the most effective cure for controlling acidosis. However, C. vulgaris and Schizochytrium spp. had an effect on medium culture pH and demonstrated potential for acidosis prevention. This study found that algae can influence ruminal fermentation, have the potential to reduce CH4 production, and may reduce acidosis incidence rates. These assumptions, however, must be validated through in vivo studies. Full article
(This article belongs to the Special Issue In Vitro Fermentation, 2nd Edition)
11 pages, 2371 KiB  
Article
Supplementing Proteolytic Enzymes Increased the In Vitro Nutrient Effective Degradability and Fermentation Characteristics of Pineapple Waste Silage
by Kim Margarette Corpuz Nogoy, Jae Ik Lee, Jia Yu, Jung In Sang, Hyoung Ki Hong, Yoon Gwang Ji, Xiang Zi Li and Seong Ho Choi
Fermentation 2023, 9(3), 218; https://doi.org/10.3390/fermentation9030218 - 24 Feb 2023
Cited by 1 | Viewed by 1404
Abstract
Pineapple waste silage (PAS) is an abundant agro-industrial by-product characterized by its high fiber content posing a high potential feed value as roughage for ruminants. Studies on its supplementation with proteolytic enzyme (PE) will help extend its utilization as an alternative nutritive feed [...] Read more.
Pineapple waste silage (PAS) is an abundant agro-industrial by-product characterized by its high fiber content posing a high potential feed value as roughage for ruminants. Studies on its supplementation with proteolytic enzyme (PE) will help extend its utilization as an alternative nutritive feed source for cattle nutrition. Thus, this study aimed to determine the in vitro nutrient degradability and fermentation characteristics of fiber-rich but low-protein PAS supplemented with different levels of PE. Seven treatments were evaluated in this study: PAS without PE and PAS1 to PAS6, which corresponds to incremental levels of PAS supplementation as follows: 0.1%, 0.2%, 0.3%, 1%, 2%, and 4%. The nutrient disappearance, nutrient effective degradability, and fermentation characteristics such as total gas production, ammonia-nitrogen, and pH values were evaluated in vitro. PAS without added PE showed a comparably good nutritive value (dry matter: 94.30%, neutral detergent fiber: 63.66%, acid detergent fiber: 34.78%) to that of commonly used corn silage in South Korea. With the supplementation of PE in PAS, the PE increased the effective degradability of different nutrients such as dry matter (DM), neutral detergent fiber (NDF), acid detergent fiber (ADF), organic matter (OM), and crude protein (CP). The effect of PE supplementation on the degradation of nutrients was consistent with improvements in in vitro rumen fermentation characteristics. Supplementing PAS with PE increased the total gas production and decreased the pH values, which are characteristics of heightened fiber degradation and fermentation. The ammonia-N concentration of the in vitro-incubated PAS was moderated by the addition of PE, which is likely due to the decrease in pH or in vitro acidosis and has shown a synergistic protease activity effect on nutrient degradation. Overall, supplementing PAS with PE increased the effective degradability of DM, NDF, ADF, OM, and CP, with the most dramatic effects observed in PAS3 and PAS6 (0.3% and 4%, respectively). Full article
(This article belongs to the Special Issue In Vitro Fermentation, 2nd Edition)
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15 pages, 1107 KiB  
Article
Seasonal Variation in Chemical Composition, Ruminal Fermentation, and Biological Characteristics of Paulownia shan tong: In Vitro Potential Use by Sheep and Goats
by Hajer Ammar, Ahmed E. Kholif, Manel Missaoui, Halimeh Zoabi, Soha Ghzayel, Mario de Haro-Martí, Izabelle Auxiliadora Molina de Almeida Teixeira, Sondos Fkiri, Mohamed Larbi Khouja, Mahmoud Fahmy, Gouda A. Gouda, Secundino López and Mireille Chahine
Fermentation 2023, 9(3), 210; https://doi.org/10.3390/fermentation9030210 - 23 Feb 2023
Cited by 1 | Viewed by 1954
Abstract
This study was conducted to determine the seasonal variation (two seasonal harvests during winter and spring) in some chemical and biochemical characteristics of Paulownia shan tong leaves. To achieve this, we analyzed the chemical composition in terms of dry matter (DM), organic matter [...] Read more.
This study was conducted to determine the seasonal variation (two seasonal harvests during winter and spring) in some chemical and biochemical characteristics of Paulownia shan tong leaves. To achieve this, we analyzed the chemical composition in terms of dry matter (DM), organic matter (OM), crude protein (CP), cell-wall content (neutral detergent fiber (NDF)), acid detergent fiber (ADF), acid detergent lignin (ADL), and lipids. The phenolic compounds (PC) and antioxidant activity (AA) of the leaves were also determined. In vitro gas production (GP) and nutrient degradability were evaluated using two ruminal-content donors (sheep and goats). Higher (p < 0.05) antioxidant activities were observed in the winter harvest of P. shan tong, while higher concentrations of OM (90.5%), lipids (3.02%), NDF (69.7%), ADF (54.8%), ADL (35.4%), total flavonoids (0.45 mg catechin equivalents/mg DM), and total phenolics (2.52 mg gallic acid equivalents/mg DM) were observed in the spring harvest. Higher asymptotic GP and rate of GP (p < 0.05) were observed when the leaves were incubated with rumen liquor of goats compared to sheep; however, higher (p < 0.05) lag time of GP and half-time of GP were observed with rumen liquor from sheep. Moreover, higher asymptotic GP, rate of GP, and half-time of GP were observed with the winter harvest of P. shan tong. Higher (p < 0.001) metabolizable energy, OM degradability, and microbial protein supplies were obtained when the leaves were incubated with rumen fluids from goats and from the winter harvest of P. shan tong compared to the spring harvest. In conclusion, based on the chemical composition and the kinetics of in vitro gas production, P. shan tong leaves seem to be characterized by a good nutritive value and could be considered as an alternative feed resource for ruminants. Full article
(This article belongs to the Special Issue In Vitro Fermentation, 2nd Edition)
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12 pages, 283 KiB  
Article
Effect of Heat Processing of Rubber Seed Kernel on In Vitro Rumen Biohydrogenation of Fatty Acids and Fermentation
by Nirawan Gunun, Metha Wanapat, Chatchai Kaewpila, Waroon Khota, Sineenart Polyorach, Anusorn Cherdthong, Rattikan Suwannasing, Pairote Patarapreecha, Piyawit Kesorn, Piyawan Intarapanich, Nuttabodee Viriyawattana and Pongsatorn Gunun
Fermentation 2023, 9(2), 143; https://doi.org/10.3390/fermentation9020143 - 01 Feb 2023
Cited by 2 | Viewed by 1622
Abstract
The aim of this study was to assess the effect of rubber seed kernel heat processing on in vitro rumen biohydrogenation of fatty acids and fermentation. The experiment was conducted with a completely randomized design (CRD). The inclusion of RSK at 0% (CON) [...] Read more.
The aim of this study was to assess the effect of rubber seed kernel heat processing on in vitro rumen biohydrogenation of fatty acids and fermentation. The experiment was conducted with a completely randomized design (CRD). The inclusion of RSK at 0% (CON) and 20% with different processing methods as follows: Raw rubber seed kernel (RAWR), roasted rubber seed kernel (ROR), microwave irradiated rubber seed kernel (MIR), and rubber seed kernel were heated in a hot air oven (RHO) in total mixed ration (TMR) diets. The hydrogen cyanide (HCN) was reduced using RSK heat methods. The heat processing of RSK had no effect on cumulative gas production at 96 h, the gas production from the insoluble fraction (b), or degradability (p > 0.05), whereas it reduced the gas production from the immediately soluble fraction (a) and constant rate of gas production for the insoluble fraction (c) (p < 0.01). The RSK processing methods did not influence ruminal pH, total volatile fatty acid (VFA), or VFA proportions (p > 0.05). RSK heat processing reduced ammonia-nitrogen (NH3-N) (p < 0.04) while increasing the bacterial population (p < 0.02). Heat treatment had no effect on linoleic acid (C18:2 cis-9,12 + tran-9,12) (p > 0.05). The RHO increases oleic acid (C18:1 cis-9 + tran-9) and linolenic acid (C18:3 cis-9,12,15) concentrations (p < 0.01). In conclusion, RHO reduced rumen biohydrogenation of unsaturated fatty acids (UFA), especially C18:3 and C18:1. Full article
(This article belongs to the Special Issue In Vitro Fermentation, 2nd Edition)
7 pages, 439 KiB  
Article
Effect of Sodium Hydroxide Treatment on Chemical Composition and Feed Value of Common Reed (Phragmites australis) Straw
by Ahmet Uzatici, Onder Canbolat and Adem Kamalak
Fermentation 2022, 8(12), 749; https://doi.org/10.3390/fermentation8120749 - 16 Dec 2022
Cited by 2 | Viewed by 1987
Abstract
This research was undertaken with the aim of determining the effect of sodium hydroxide (NaOH) treatment on the chemical composition, in vitro gas production, neutral detergent fiber digestibility (NDFD) and true dry matter digestibility (TDMD), dry matter intake (DMI), and relative feed value [...] Read more.
This research was undertaken with the aim of determining the effect of sodium hydroxide (NaOH) treatment on the chemical composition, in vitro gas production, neutral detergent fiber digestibility (NDFD) and true dry matter digestibility (TDMD), dry matter intake (DMI), and relative feed value (RFV) of common reed (Phragmites australis) straw. Reed straw was treated with 0% (control), 1%, 2%, and 3% NaOH and stored in 1.5-L glass jars in triplicate for 21 days. NaOH treatment had a significant effect on the chemical composition, in vitro gas production, NDFD and TDMD, DMI, and RFV of the reed straw. While it decreased the cell wall content of the reed straw, it significantly increased the NDFD, TDMD, DMI, and RFV. The neutral detergent fiber content of the reed straw decreased with NaOH treatment in a dose-dependent manner and ranged from 56.03% to 65.05%, whereas the NDFD increased and ranged from 53.10% to 59.99%. Metabolizable energy, organic matter digestibility, and TDMD values were improved, ranging from 9.15 to 10.19 MJ/kg DM, 58.46% to 65.05%, and 55.29% to 62.33%, respectively. The estimated RFV and DMI also improved, ranging from 84.70% to 95.58% and from 1.87% to 2.14% of body weight, respectively. The most effective treatment dose of NaOH was 3%. Therefore, it can be suggested that NaOH treatment has the potential to improve the nutritive value of reed straw. However, before large applications, further in vivo investigations are required to determine the effects of NaOH treatment on the feed intake and production of ruminant animals. Full article
(This article belongs to the Special Issue In Vitro Fermentation, 2nd Edition)
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11 pages, 271 KiB  
Article
Effect of Source and Level of Dietary Supplementary Copper on In Vitro Rumen Fermentation in Growing Yaks
by Xinsheng Zhao, Lizhuang Hao, Yanfeng Xue, Allan Degen and Shujie Liu
Fermentation 2022, 8(12), 693; https://doi.org/10.3390/fermentation8120693 - 30 Nov 2022
Cited by 3 | Viewed by 1452
Abstract
Copper (Cu) is essential for the health of livestock, however, the optimal source and level of dietary Cu for yaks are uncertain. To fill this important gap, we designed an in vitro study to examine the effects of three Cu sources, namely Cu [...] Read more.
Copper (Cu) is essential for the health of livestock, however, the optimal source and level of dietary Cu for yaks are uncertain. To fill this important gap, we designed an in vitro study to examine the effects of three Cu sources, namely Cu methionine (Met-Cu), Cu chloride (CuCl2) and tribasic Cu chloride (TBCC), at five levels, namely 5, 10, 15, 20 and 25 mg/kg DM (includes Cu in substrate), on rumen fermentation in yaks. In vitro dry matter degradability (IVDMD) and amylase activity were greater (p < 0.05) with added Met-Cu than the other two Cu sources, and ammonia nitrogen (NH3-N), microbial protein (MCP) and propionate contents were greater with Met-Cu and CuCl2 than with TBCC. Total gas production and lipase activity were greater with Met-Cu and TBCC than CuCl2 (p < 0.05), which meant that the metabolizable energy yield was greater in the two former Cu sources than the latter, but CH4 production did not differ (p = 0.92) among Cu sources. IVDMD and lipase activity were greatest (p < 0.05) at 15 mg Cu/kg DM in the substrate and MCP, isobutyrate, butyrate and isovalerate contents, and amylase and trypsin activities were greatest or second greatest at 10 and 15 mg Cu/kg DM. It was concluded that Met-Cu was the best source of Cu and 10 to 15 mg Cu/kg DM was the optimal level for yaks, at least under in vitro conditions. Full article
(This article belongs to the Special Issue In Vitro Fermentation, 2nd Edition)
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