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18 pages, 896 KiB

Open AccessArticle

Influence of Genotype and Anaerobic Fermentation on In Vitro Rumen Fermentation Characteristics and Greenhouse Gas Production of Whole-Plant Maize

by Edwin Rafael Alvarado-Ramírez, Mona Mohamed Mohamed Yasseen Elghandour, Marco Antonio Rivas-Jacobo, Serena Calabrò, Alessandro Vastolo, Monica Isabella Cutrignelli, Pedro Enrique Hernández-Ruiz, Edson Brodeli Figueroa-Pacheco and Abdelfattah Zeidan Mohamed Salem

Fermentation 2024, 10(1), 42; https://doi.org/10.3390/fermentation10010042 - 04 Jan 2024

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Abstract

The objective of this study was to evaluate the effect of anaerobic fermentation on the in vitro ruminal production of total gas (TG), methane (CH₄), carbon monoxide (CO) and hydrogen sulfide (H₂S), as well as on the characteristics of [...] Read more.

The objective of this study was to evaluate the effect of anaerobic fermentation on the in vitro ruminal production of total gas (TG), methane (CH₄), carbon monoxide (CO) and hydrogen sulfide (H₂S), as well as on the characteristics of ruminal fermentation and CH₄ conversion efficiency of whole-plant maize (WPM) from four native maize genotypes (Amarillo, Olotillo, Tampiqueño and Tuxpeño) from Mexico, and a commercial hybrid was used as a control. In all genotypes, the fermented WPM produced the lowest amounts (p ≤ 0.0236) of TG and CH₄ from degraded dry matter (DM), and Tampiqueño and Tuxpeño presented the highest production of these gases. In addition, Tuxpeño also presented the highest proportion of CH₄ (mL 100 mL⁻¹ TG), and Olotillo presented the lowest proportion of both gases. Something similar occurred in H₂S, where the fermented WPM produced the lowest (p ≤ 0.0077) amount per DM degraded, and Amarillo and Tampiqueño presented the highest and lowest production, respectively. However, the fermented WPM presented the highest (p = 0.0128) CO production from degraded DM, and Tuxpeño and Olotillo presented the highest and lowest production, while the rumen pH was lower (p < 0.0001) in the fermented WPM, and Tuxpeño and Olotillo presented the highest and lowest pH, respectively. Furthermore, the fermented plant presented the greatest (p ≤ 0.0055) DM degradation, and the Amarillo and hybrid genotypes presented the highest percentages, while Olotillo presented the lowest. The short-chain fatty acid (SCFA) content and metabolizable energy (ME) did not differ (p ≥ 0.0899) between genotypes and were higher (p = 0.0009) in the fresh WPM. Despite the above, the fermented WPM was more efficient (p ≤ 0.0249), and the Amarillo and hybrid genotypes produced less CH₄ per unit of SCFAs, ME and organic matter. In conclusion, the Amarillo genotype equaled the hybrid one, and although the production of CO increased, anaerobic fermentation showed the potential to reduce the rumen production of TG, CH₄ and H₂S, as well as to improve DM degradability and CH₄ conversion efficiency. Full article

(This article belongs to the Special Issue In Vitro Fermentation, 3rd Edition)

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21 pages, 2919 KiB

Open AccessArticle

A Comparison of Three Artificial Rumen Systems for Rumen Microbiome Modeling

by Claire A. Shaw, Yuna Park, Maria Gonzalez, Rich A. Duong, Pramod K. Pandey, Charles G. Brooke and Matthias Hess

Fermentation 2023, 9(11), 953; https://doi.org/10.3390/fermentation9110953 - 07 Nov 2023

Cited by 1 | Viewed by 1861

Abstract

The rumen contains a complex mixture of microbes, crucial for the animal’s ability to degrade feed. Some of the feed-derived carbon is released as methane, a potent greenhouse gas, into the atmosphere. There is growing interest in reducing the loss of feed-derived carbon, [...] Read more.

The rumen contains a complex mixture of microbes, crucial for the animal’s ability to degrade feed. Some of the feed-derived carbon is released as methane, a potent greenhouse gas, into the atmosphere. There is growing interest in reducing the loss of feed-derived carbon, making it available to the animal and improving animal productivity. Artificial rumen systems (ARSs) have been widely used to evaluate novel feed additives in terms of their ability to reduce methane production in the rumen and their effect on the rumen microbiome function prior to conducting resource-intensive animal trials. While the value of ARSs is widely acknowledged, it remains unclear which of these in vitro systems simulate the natural system most accurately. Here, we evaluated three different ARSs and compared them to in vivo rumen metrics. The results showed that all systems were capable of maintaining stable pH, redox potential, and temperature over time. The batch-style ARS simulated the rumen over 48 h. The semi-continuous ARS mimicked the volatile fatty acid profile and microbiota of the in vivo rumen for up to 120 h. Similarly, all ARSs maintained the prokaryotic and eukaryotic rumen populations over the duration of the study, with the semi-continuous ARS maintaining the natural rumen microbiome more accurately and for up to 120 h. In sum, our results suggest that three of the widely used ARSs simulate the rumen ecosystem adequately for many short-term rumen microbiome studies, with the more advanced semi-continuous ARS being more accurate when rumen simulation is extended to over 48 h. Full article

(This article belongs to the Special Issue In Vitro Fermentation, 3rd Edition)

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18 pages, 1771 KiB

Open AccessArticle

Ex Vivo Fermentation of Hay and Corn by Rumen Bacteria from Cattle and Sheep

by Jennifer R. Weinert-Nelson, Donald G. Ely, Michael D. Flythe, Tracy A. Hamilton, Jessica L. Ferrell and Brittany E. Davis

Fermentation 2023, 9(11), 929; https://doi.org/10.3390/fermentation9110929 - 25 Oct 2023

Cited by 1 | Viewed by 995

Abstract

Sheep are often utilized as a model ruminant, despite a lack of functional comparisons of rumen bacterial communities and responses during dietary transitions between sheep and cattle. Therefore, an ex vivo study was conducted to evaluate species differences. Rumen fluid was obtained from [...] Read more.

Sheep are often utilized as a model ruminant, despite a lack of functional comparisons of rumen bacterial communities and responses during dietary transitions between sheep and cattle. Therefore, an ex vivo study was conducted to evaluate species differences. Rumen fluid was obtained from hay-fed sheep and cattle (n = 3 species⁻¹). Mixed bacterial cell suspensions in buffered media containing 3% w/v ground hay, corn, or combinations (2:1, 1:2) of substrates were incubated (24 h; 39 °C). Suspension pH, lactate, volatile fatty acids (VFA), and digestibility were assessed, functional guilds enumerated, and amylolytic bacteria isolated. Lactate was fully utilized in all hay incubations, and pH did not differ between species (p > 0.75). In contrast, digestibility, lactate accumulation, and pH decline were greater in bovine suspensions fermenting corn (p < 0.01). Streptococcus bovis was the predominant bacteria regardless of species, but total amylolytic bacteria were 10-fold greater in bovine suspensions (p < 0.01). Lactate-utilizing bacteria were 1000-fold greater in bovine than ovine suspensions (p < 0.01). However, total VFA did not differ between species (p > 0.28). Overall, these results demonstrate differential feed utilization capacities in rumen microbial communities of sheep and cattle as well as potential differences in rumen acidosis susceptibility. Full article

(This article belongs to the Special Issue In Vitro Fermentation, 3rd Edition)

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16 pages, 2272 KiB

Open AccessArticle

Fructan Catabolism by Rumen Microbiota of Cattle and Sheep

by Jennifer R. Weinert-Nelson, Isabelle A. Kagan, Donald G. Ely, Michael D. Flythe and Brittany E. Davis

Fermentation 2023, 9(11), 925; https://doi.org/10.3390/fermentation9110925 - 24 Oct 2023

Viewed by 916

Abstract

Fructans serve as the primary form of storage carbohydrate in cool-season grasses, but little is known about potential differences in ruminal fermentation of fructans between cattle and sheep. An ex vivo study was conducted to evaluate species differences in fructan catabolism. Buffered media [...] Read more.

Fructans serve as the primary form of storage carbohydrate in cool-season grasses, but little is known about potential differences in ruminal fermentation of fructans between cattle and sheep. An ex vivo study was conducted to evaluate species differences in fructan catabolism. Buffered media containing ground orchardgrass (Dactylis glomerata L.) substrate was inoculated with uncultivated rumen microbiota obtained from cattle and sheep (n = 4 species⁻¹). Fructan profiles were monitored over the incubation period (8 h; 39 °C) using high-performance anion-exchange chromatography coupled to pulsed amperometric detection (HPAEC-PAD). In both species, disappearance of long-chain fructans (degree of polymerization [DP] > 8) was evident by 2 h of incubation (p < 0.01), whereas short-chain fructans (DP 4–8) increased from 0 to 2 h prior to subsequent degradation (p < 0.01). However, the overall rate of long-chain fructan catabolism was greater in bovine versus ovine fermentations, particularly between 2 and 4 h (p < 0.01). Additionally, rapid utilization of short-chain fructans occurred from 2 to 4 h in bovine fermentations, but was delayed in ovine fermentations, with substantial degradation occurring only after 4 h of incubation (p < 0.01). These results indicate that rumen microbiota of cattle may have a greater capacity for fructan degradation. Full article

(This article belongs to the Special Issue In Vitro Fermentation, 3rd Edition)

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14 pages, 3988 KiB

Open AccessArticle

Effects of Exogenous Oral Infusion of Volatile Fatty Acids on Ileal Microbiome Profiling and Epithelial Health in Goats

by Yongkang Zhen, Chong Zhang, Jiaqi Lin, Ali Rahmat, Feiyang He and Mengzhi Wang

Fermentation 2023, 9(9), 801; https://doi.org/10.3390/fermentation9090801 - 30 Aug 2023

Viewed by 737

Abstract

The role of volatile fatty acids (VFAs) in ruminal fermentation is well studied, but their effects on the ileal microbiome and epithelial health remain less explored. In this study, we investigated the impact of the exogenous oral infusion of three VFAs, sodium acetate [...] Read more.

The role of volatile fatty acids (VFAs) in ruminal fermentation is well studied, but their effects on the ileal microbiome and epithelial health remain less explored. In this study, we investigated the impact of the exogenous oral infusion of three VFAs, sodium acetate (NaAc), propionate (NaPr), and butyrate (NaBu), on the VFA concentrations in ileal contents, as well as ileal microbiome profiling and epithelial health parameters (inflammatory cytokine and tight junctions) in goats. The data demonstrated that the oral infusion of three VFAs can enhance VFA production by increasing the proportions of each individual VFA and the total VFAs. Then, the microbiome revealed distinct microbial succession patterns and altered microbial diversities in response to the oral infusion of VFA treatments. Moreover, the oral infusion of each VFA had unique effects on the ileal bacterial community, with potential implications for epithelial health. Notably, the oral infusion of VFAs demonstrated potential anti-inflammatory effects, as evidenced by reduced IL-6 levels in the NaPr group and increased IL-10 levels in the NaAc group. Notably, the oral infusion of VFAs did not directly impact the tight junction concentrations, such as Claudin1, Occludin, and ZO-1. Lastly, the correlation analyses identified specific relationships between the ileal bacteria and epithelial health parameters, and Prevotella was positively correlated with IL-6 and IL-1β, while Bifidobacterium was positively correlated with IL-10. These results highlighted the crosstalk between VFAs, the ileal microbiome, and the health of the small intestine. Our findings had significant implications for optimizing ruminant nutrition, enhancing epithelial health, and promoting sustainable livestock production practices. Full article

(This article belongs to the Special Issue In Vitro Fermentation, 3rd Edition)

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13 pages, 3147 KiB

Open AccessArticle

Effects of Oligosaccharide Fermentation on Canine Gut Microbiota and Fermentation Metabolites in an In Vitro Fecal Fermentation Model

by Yu Zhang, Zhuang Ding, Xiaoyu Chen, Min Wen, Qingpeng Wang and Zhengping Wang

Fermentation 2023, 9(8), 722; https://doi.org/10.3390/fermentation9080722 - 01 Aug 2023

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Abstract

Owing to the well-established application of prebiotics in human food products, there is a growing interest in their potential as dietary supplements for gut microbiota composition and improvement of the digestive health of dogs. However, targeted studies with dogs as research subjects are [...] Read more.

Owing to the well-established application of prebiotics in human food products, there is a growing interest in their potential as dietary supplements for gut microbiota composition and improvement of the digestive health of dogs. However, targeted studies with dogs as research subjects are still limited. In the present study, an in vitro simulated gut microbiota fermentation system using canine feces from a healthy Border Collie breed was used to investigate the prebiotic effects of five different oligosaccharides and compare their regulatory effects on the gut microbiota structure and the resultant metabolites. Due to the addition of oligosaccharides, the fermented samples had lower pH and higher bacterial proliferation. The oligosaccharide-fermentation selectively boosted Lactobacillus spp., Streptococcus spp., Enterococcus spp., Bacteroides spp., and hindered Escherichia-Shigella spp., Paeniclostridium, spp., and Bacteroides spp. Each oligosaccharide showed distinct characteristics and preferences for regulating gut microbiota structure and abundance. Furthermore, the addition of oligosaccharides increased the production of short-chain fatty acids, particularly butyric acid. This study provides a preliminary basis for the rapid and rational selection of prebiotic oligosaccharides as canine dietary supplements and further explores the function of oligosaccharides and their combinations in canine health. Full article

(This article belongs to the Special Issue In Vitro Fermentation, 3rd Edition)

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20 pages, 3494 KiB

Open AccessArticle

Effects of Administration of Prebiotics Alone or in Combination with Probiotics on In Vitro Fermentation Kinetics, Malodor Compound Emission and Microbial Community Structure in Swine

by Maro Lee, Yeonjae Choi, Joel Bayo, Andrew Wange Bugenyi, Yangseon Kim and Jaeyoung Heo

Fermentation 2023, 9(8), 716; https://doi.org/10.3390/fermentation9080716 - 28 Jul 2023

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Abstract

The objective of this study was to evaluate the effect of Lactobacillus amylovorus, L. plantarum, galacto-oligosaccharide (GOS) and their synbiotic formulations on pH, volatile fatty acids (VFA), malodor, and microbial ecological profiles through a 24-h in vitro fermentation model. Inclusion of [...] Read more.

The objective of this study was to evaluate the effect of Lactobacillus amylovorus, L. plantarum, galacto-oligosaccharide (GOS) and their synbiotic formulations on pH, volatile fatty acids (VFA), malodor, and microbial ecological profiles through a 24-h in vitro fermentation model. Inclusion of GOS alone and in synbiotic combination with either probiotic resulted in consistently lower pH and higher total gas volumes at 12 and 24 h of incubation. Notably, concentrations of odorous compounds (hydrogen sulfide, H₂S and methyl mercaptan, CH₃SH) in the total gas produced were significantly lower in these GOS-containing treatments relative to the controls and probiotic-only-treated groups. However, although ammonia showed an initial relative reduction at 12 h, concentrations did not differ among treatments at 24 h. Further, the GOS-containing treatments had remarkably higher total and individual VFAs, including acetate, propionate, and butyrate, relative to controls and the probiotic-only treatments. Analysis of microbial composition and diversity showed clustering of GOS-containing treatments away from the controls and probiotic-only treatments at 12 and 24 h of incubation. Our study suggests that GOS supplementation (alone or in combination with L. amylovorus or L. plantarum probiotic strains) has the potential to increase VFA production in the swine gut while lowering emissions of malodorous compounds, except ammonia, in their manure. Full article

(This article belongs to the Special Issue In Vitro Fermentation, 3rd Edition)

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15 pages, 658 KiB

Open AccessArticle

Dry Matter Content and Additives with Different Modes of Action Modify the Preservation Characteristics of Grass Silage

by Marcia Franco and Marketta Rinne

Fermentation 2023, 9(7), 640; https://doi.org/10.3390/fermentation9070640 - 08 Jul 2023

Cited by 2 | Viewed by 1065

Abstract

Two experiments evaluated how grass silage quality can be manipulated by various management options. In Experiment 1, silage characteristics were evaluated at two dry matter (DM) contents and treated with additives presenting different modes of action. Timothy grass was ensiled at low (224 [...] Read more.

Two experiments evaluated how grass silage quality can be manipulated by various management options. In Experiment 1, silage characteristics were evaluated at two dry matter (DM) contents and treated with additives presenting different modes of action. Timothy grass was ensiled at low (224 g/kg) and high (534 g/kg) DM contents and five additives were applied: 1. control (C), 2. homofermentative lactic acid bacteria inoculant (HO), 3. heterofermentative lactic acid bacteria inoculant (HE), 4. salt-based additive (SA) and 5. Formic- and propionic-acid-based additive (FPA). A higher DM content and FPA restricted silage fermentation, and additive effects were generally greater in low rather than high DM silages. The chemical additives SA and FPA resulted in the highest aerobic stability, while the HE improved it at a high DM content. In Experiment 2, the low DM content grass was ensiled utilizing resin acids, as follows: 1. C, 2. FPA, 3. Resin acid oil (FOR) at 13 L/t, 4. FOR at 26 L/t, 5. Resin acid soluble in water (ROS) at 13 L/t and 6. ROS at 26 L/t. Wilting combined with additives improved the preservation characteristics of grass silages (Experiment 1), but resin acid products failed to modify silage fermentation compared to the control (Experiment 2). Full article

(This article belongs to the Special Issue In Vitro Fermentation, 3rd Edition)

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19 pages, 5807 KiB

Open AccessArticle

16S rRNA Sequencing Reveals the Antibacterial Effect of Omega-3 (Fish Oil) against Fibrolytic Bacteria, Altering Fermentation and Volatile Fatty Acids Profile In Vitro

by Mohamed Abdelrahman, Wei Wang, Zhigao An, Haimiao Lv, Guohua Hua, Ahmed Ezzat Ahmed, Aiman Alsaegh and Liguo Yang

Fermentation 2023, 9(7), 596; https://doi.org/10.3390/fermentation9070596 - 26 Jun 2023

Cited by 1 | Viewed by 1131

Abstract

Polyunsaturated fatty acids (PUFAs) constitute a significant lipid class with essential nutritional and health benefits for both animal and human health; however, their effect and interaction with the gut microbiota ecosystem are still unclear. Therefore, the present study aims to investigate the effect [...] Read more.

Polyunsaturated fatty acids (PUFAs) constitute a significant lipid class with essential nutritional and health benefits for both animal and human health; however, their effect and interaction with the gut microbiota ecosystem are still unclear. Therefore, the present study aims to investigate the effect of fish oil (FO) on ruminal fermentation and bacterial abundance under high- and low-forage diets. Thirty-six ruminal fluid samples were allocated into two experiments. The first was on high-forage diet and included three groups: the control (basal diet with 70% forage and 30% concentrate), group 2 (basal diet + 5 mL/L FO), and group 3 (basal diet + 10 mL/L). The second experiment was on low-forage diet: the control (basal diet with 30% forage and 70% concentrate), group 2 (basal diet + 5 mL/L FO), and group 3 (basal diet + 10 mL/L). The results showed that although FO supplementation did not affect the pH level among different diets, it significantly decreased methane under a high-forage diet. In addition, regarding the fatty acids profile, FO supplementation in high-forage diet significantly decreased fatty acids in both; however, under a low-forage diet, FO groups showed significantly higher fatty acid content than the control. However, FO supplementation increased the abundance of Anaerovibirio, Selenomonas, pseudobutyrivibrio, and butyrivibrio through a high-forage diet. In contrast, the abundance of Prevotella, Rikenellaceae RC9 gut group, and Saccharofermentans was depressed with FO supplementation. Whereas under low-forage diet, FO supplementation increased Ruminobacter, Anaerovibirio, Megasphaera, Pseudobutyrivibrio, Streptococcus, Butyrivibrio, unclassified_lachnospiraceae; it also decreased Prevotella and Rikenellaceae RC9 abundance similar to the high-forage diet. Based on the KEGG pathway results, FO supplementation significantly downregulated genes mainly related to folding, sorting and degradation, environmental adaptation, cell motility, transcription, membrane transport, and signal transduction. The results revealed that FO has a depressing effect on ruminal fermentation and some bacterial population; however, this negative effect can be minimized in high-concentrate diets. Full article

(This article belongs to the Special Issue In Vitro Fermentation, 3rd Edition)

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14 pages, 2602 KiB

Open AccessArticle

Calcium Propionate Supplementation Mitigated Adverse Effects of Incubation Temperature Shift on In Vitro Fermentation by Modulating Microbial Composition

by Tengfei He, Xilin Wang, Shenfei Long, Jiangong Li, Zhenlong Wu, Yao Guo, Fang Sun and Zhaohui Chen

Fermentation 2023, 9(6), 544; https://doi.org/10.3390/fermentation9060544 - 06 Jun 2023

Cited by 2 | Viewed by 1216

Abstract

The ingestion of cold water in winter results in fluctuating decreases in rumen temperature, impacting rumen fermentation function and methane production. This study aimed to investigate the impact of calcium propionate (CaP) on rumen fermentation characteristics, methane production, and microbial microbiome under different [...] Read more.

The ingestion of cold water in winter results in fluctuating decreases in rumen temperature, impacting rumen fermentation function and methane production. This study aimed to investigate the impact of calcium propionate (CaP) on rumen fermentation characteristics, methane production, and microbial microbiome under different in vitro incubation temperatures. A 2 × 2 factorial experiment was conducted over 72 h, with or without 2.5% CaP (dry matter [DM] basis) in substrate under a constant incubation temperature (39 °C, CIT) or shifting incubation temperatures (12 cycles in total, with each cycle consisting of 4 h of incubation at 39 °C followed by immediate transfer to 30 °C for 2 h, ITS). The results showed that ITS inhibited the gas production, methane production, and methane concentration at 12 and 72 h (p < 0.05), and reduced the concentration of ammonium nitrogen (−14.25%), propionate (−16.14%), butyrate (−12.67%), and total volatile fatty acid (−8.50%) at 72 h more than the CIT groups (p < 0.05). The addition of 2.5% CaP significantly increased the gas production at 72 h (+4.84%), asymptotic gas production (+5.08%, per 0.2 g DM substrate), concentration of propionate (+18.05%), and valerate (+9.45%) (p < 0.05) compared to CaP-free groups, while it had no observable effect on the production or concentration of methane. Furthermore, the addition of 2.5% CaP yielded a significant increase in the relative abundance of Bacteroides (p < 0.05). Under the ITS condition, the relative abundance of Methanomicrobiales decreased and was positively correlated with methane production at 72 h (r = 0.47, p < 0.05). Additionally, CaP decreased the abundance of Prevotella_UCG_003 (p < 0.05), which was negatively correlated with the asymptotic gas production (r = 0.45, p < 0.05). Overall, our study suggests that the addition of 2.5% CaP can alleviate the adverse effects of ITS on in vitro fermentation parameters by regulating microbial composition and sustaining a reduction in methane production. Full article

(This article belongs to the Special Issue In Vitro Fermentation, 3rd Edition)

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Review

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19 pages, 291 KiB

Open AccessReview

Enhancement of the Nutritional Composition and Antioxidant Activities of Fruit Pomaces and Agro-Industrial Byproducts through Solid-State Fermentation for Livestock Nutrition: A Review

by Olusegun Oyebade Ikusika, Oluwakamisi Festus Akinmoladun and Conference Thando Mpendulo

Fermentation 2024, 10(5), 227; https://doi.org/10.3390/fermentation10050227 - 25 Apr 2024

Viewed by 259

Abstract

The abundance of fruit waste from the food industry and wineries, particularly peels, seeds, and other fruit pomace throughout the year, could lead to health and environmental hazards if not channelled into productive areas. Improving or transforming these waste products for better use [...] Read more.

The abundance of fruit waste from the food industry and wineries, particularly peels, seeds, and other fruit pomace throughout the year, could lead to health and environmental hazards if not channelled into productive areas. Improving or transforming these waste products for better use in other vital sectors could be achieved via solid-state fermentation (SSF) since most waste products are solid. One such productive and important area is the feeding of livestock, which will guarantee millennium food security goals for many nations of the world. The nutritional and antioxidant composition of abundantly available fruit pomace and agro-industrial byproducts could be improved via solid-state fermentation for overall livestock productivity. They contain substantial dietary fibre, protein, and phenolic compounds; hence, improving them via fermentation could serve the livestock industry in dual capacities, including nutraceutical and conventional feedstuff. This review seeks to provide reinforcing evidence on the applicability and impact of fruit pomaces on livestock nutrition. The significant nutrient improvements, beneficial outcomes in feeding trials, and inconsistencies or areas of research gap were also explored. Full article

(This article belongs to the Special Issue In Vitro Fermentation, 3rd Edition)

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