Microbial Protein and Metabolite Profiles of Klebsiella oxytoca M5A1 in a Bubble Column Bioreactor.

The production of microbial proteins (MPs) has emerged as a critical focus in biotechnology, driven by the need for sustainable and scalable alternatives to traditional protein sources. This study investigates the efficacy of two experimental setups in producing MPs using the nitrogen-fixing bacterium Klebsiella oxytoca M5A1. K. oxytoca M5A1, known for its facultative anaerobic growth and capability to fix atmospheric nitrogen, offers a promising avenue for environmentally friendly protein production. This research compares the performance of a simple bubble column (BC) bioreactor, which promotes efficient mixing and cross-membrane gas transfer, with static fermentation, a traditional method lacking agitation and aeration. The study involved the parallel cultivation of K. oxytoca M5A1 in both systems, with key parameters such as microbial growth, glucose utilization, protein concentration, and metabolite profiles monitored over a 48 h period. The results indicate that the BC bioreactor consistently outperformed static fermentation regarding the growth rate, protein yield, and glucose utilization efficiency. The BC exhibited a significant increase in protein production, reaching 299.90 µg/mL at 48 h, compared to 219.44 µg/mL in static fermentation. The organic acid profile reveals both synthesis and utilization regimes of varying patterns. These findings highlight the advantages of the BC bioreactor for MP production, particularly its ability to maintain aerobic conditions that support higher growth and yield.

Sustainable production of microbial protein from carbon dioxide in the integrated bioelectrochemical system using recycled nitrogen sources.

Given the urgency of climate change, it is imperative to develop innovative technologies for repurposing CO2 into value-added products to achieve carbon neutrality. Additionally, repurposing nitrogen-source-derived wastewater streams is crucial, focusing on sustainability rather than conventional nitrogen removal in wastewater treatment plants. In this context, microbial protein (MP) production presents a sustainable and promising approach for transforming recovered low-value resources into high-quality feed and food. We assessed MP production by hydrogen-oxidizing bacteria (HOB) utilizing CO2 and various nitrogen sources. Specifically, we investigated MP production by two different HOB strains, Cupriavidus necator H16 and Xanthobacter viscosus 7d, within an integrated water-splitting biosynthetic system that generates in situ H2 via water electrolysis. The electroautotrophically produced MPs of C. necator H16 and X. viscosus 7d exhibited amino acid contents of 555 and 717 mg protein/g cell dry weight, with 243 and 299 mg essential amino acid/g cell dry weight, respectively. They could serve as viable alternatives to conventional food/feed sources like fishmeal or soybean protein. Ammonium-rich wastewater streams are preferable for MP production in integrated bioelectrochemical systems. This study provides valuable insights into sustainable, carbon-neutral MP production using CO2, water, renewable electricity, and recycled nitrogen sources.

Power-to-vitamins: producing folate (vitamin B9) from renewable electric power and CO2 with a microbial protein system.

We recently proposed a two-stage Power-to-Protein technology to produce microbial protein from renewable electric power and CO2. Two stages were operated in series: Clostridium ljungdahlii in Stage A to reduce CO2 with H2 into acetate, and Saccharomyces cerevisiae in Stage B to utilize O2 and produce microbial protein from acetate. Renewable energy can be used to power water electrolysis to produce H2 and O2. A drawback of Stage A was the need for continuous vitamin supplementation. In this study, by using the more robust thermophilic acetogen Thermoanaerobacter kivui instead of C. ljungdahlii, vitamin supplementation was no longer needed. Additionally, S. cerevisiae produced folate when grown with acetate as a sole carbon source, achieving a total folate concentration of 6.7 mg per 100 g biomass with an average biomass concentration of 3 g l-1. The developed Power-to-Vitamin system enables folate production from renewable power and CO2 with zero or negative net-carbon emissions.

Upcycling of nutrients from kitchen waste: Integration of anaerobic digestion system and microbial protein production system.

To upcycle the nutrients from kitchen waste (KW), an integrated system consisting of anaerobic digestion (AD) reactor and microbial protein (MP) production reactor was established in this study. The subsystem I (AD system) demonstrated an efficient bio-energy production (282.37 mL CH4/g VS), with 553.54 mg/L of NH4+-N remained in the digestate. The subsystem II (MP production system) utilized the nitrogenous constituents of the digestate, with 2.04 g/L MP production. In order to further enhance the recovery efficiency, C/N ratio in the subsystem II was studied. NH4+-N recovery efficiency was 23.08% higher after C/N ratio optimization along with 0.24 g/L increment on MP production. Over 0.7 g/L of essential amino acids was obtained, according with the qualitative necessary for the feeds. Also, the key enzyme abundance of CO2 releasing and amino acid biosynthesis was obviously increased with max. 55.21%. Meanwhile, the integrated system was profitable via a simplified economic assessment.

Comparing feces collection methods for evaluating the apparent digestibility coefficient of brewers' spent yeast in juvenile Atlantic salmon (Salmo salar).

Brewer's spent yeast (BSY), derived from Saccharomyces cerevisiae used in beer production, is a valuable protein source for aquafeeds. Estimations of apparent digestibility coefficients (ADC) for nutrients in BSY are crucial for its inclusion in aquafeeds. ADC estimations for Saccharomyces cerevisiae protein in rainbow are hardly comparable from a methodological point of view, whereas the ADC estimations for BSY protein in Atlantic salmon are only based on stripped feces, which are known to produce underestimations. Therefore, new determinations of ADC of BSY nutrients are necessary for the inclusion of this ingredient in practical diets for salmonids. This study is focused on determining unbiased ADC values for protein and energy from BSY in juvenile Salmo salar. To reduce systematic biases, fecal samples were collected using stripping and decantation methods, which are known to produce under-and overestimations, respectively. 780 fish (25.16 ± 4.88 g) were stocked in six tanks. A reference diet (50% protein, 20% lipid, 1% Cr2O3) was provided to three tanks, and a test diet (70,30 reference diet to BSY) to the other three. ADC for BSY protein was 84.70 ± 1.04% (decantation) and 70.50 ± 4.03% (stripping). For gross energy, stripped feces yielded an ADC of 52.04 ± 5.30%, while decantation resulted in 63.80 ± 1.17%. Thus, ADC estimates were taken as the average of the stripping-value and the decantation-value, resulting in 77.6% for BSY crude protein, which is appreciably higher than previously measured values in S. salar fed undisrupted S. cerevisiae, and in 57.9% for gross energy.

90-day oral toxicity study in rats of a protein-rich powder derived from Xanthobacter sp. SoF1.

Xanthobacter sp. SoF1 (SoF1) is an autotrophic hydrogen-oxidizing bacteria that produces protein-rich biomass and has potential to be an alternative protein source that is more environmentally sustainable than animal and plant derived proteins. A protein-rich powder derived from SoF1 was the test material in a 90-day repeated-dose oral toxicity study to explore major toxic effects, demonstrate target organs, and provide an estimate of a no-observed-adverse-effect level (NOAEL). Daily doses of 0 (vehicle only), 375, 750, and 1500 mg/kg bw/day of the test material were administered by gavage to 10 Han:WIST rats/sex/group. An additional group was administered 1290 mg/kg bw/day whey protein concentrate as positive control. No treatment-related adverse effects were observed, and no target organs were determined after 90/91 days of consecutive administration of the test item. A NOAEL of 1500 mg/kg bw/day was determined.

Waste to protein: A systematic review of a century of advancement in microbial fermentation of agro-industrial byproducts.

Increasing global consumption of protein over the last five decades, coupled with concerns about the impact on emissions of animal-based protein production, has created interest in alternative protein sources. Microbial proteins (MPs), derived through the fermentation of agro-industrial byproducts, present a promising option. This review assesses a century of advancements in this domain. We conducted a comprehensive review and meta-analysis, examining 347 relevant research papers to identify trends, technological advancements, and key influencing factors in the production of MP. The analysis covered the types of feedstocks and microbes, fermentation methods, and the implications of nucleic acid content on the food-grade quality of proteins. A conditional inference tree model and Bayesian factor were used to ascertain the impact of various parameters on protein content. Out of all the studied parameters, such as type of feedstock (lignocellulose, free sugars, gases, and others), type of fermentation (solid, liquid, gas), type of microbe (bacteria, fungi, yeast, and mix), and operating parameters (temperature, time, and pH), the type of fermentation and microbe were identified as the largest influences on protein content. Gas and liquid fermentation demonstrated higher protein content, averaging 52% and 42%, respectively. Among microbes, bacterial species produced a higher protein content of 51%. The suitable operating parameters, such as pH, time, and temperature, were also identified for different microbes. The results point to opportunities for continued innovation in feedstock, microbes, and regulatory alignment to fully realize the potential of MP in contributing to global food security and sustainability goals.

Effects of replacing of alfalfa hay with Plantago lanceolata hay on digestibility, methane production and microbial protein production of total mixed diet.

The aim of current experiment was to determine the effect of replacement of alfalfa hay with ribwort plantain (Plantago lanceolata) hay in ruminant diets on the fermentation parameters such as gas production, methane (CH4) production, true digestible dry matter (TDDM), true digestibility (TD), partitioning factor, microbial protein, and efficiency of microbial protein using in vitro gas production technique. The alfalfa hay was replaced with P. lanceolata hay in a diets isocaloric (2650 kcal/kg DM) and nitrogenic (17% CP kg DM) at the ratio of 0, 5, 10 and 15%. Partial substitution of alfalfa hay with P. lanceolata hay had no significant effect on gas and methane (ml/incubated substrate or %) production whereas the partial substitution had a significant effect on TDDM, TD, gas (ml/digested DM), CH4 (ml ml/digested DM) and microbial MP of diets. The replacement of alfalfa hay with ribwort plantain hay shifted the fermentation pattern from gas and methane production to microbial protein production. Therefore alfalfa hay can be replaced with ribwort plantain hay with high digestibility and anti-methanogenic potential in ruminant diets up to 15% to decrease methane production and improve microbial protein production. However further in vivo experiments are required to determine the effect of replacement on feed intake and animal production.

In vitro genotoxicological evaluation of protein-rich powder derived from Xanthobacter sp. SoF1.

One way of limiting the environmental impact of food production and improving food security is to replace part of the animal- or plant-based protein in the human diet with protein sourced from microorganisms. The recently discovered bacterium Xanthobacter sp. SoF1 (VTT-E-193585) grows autotrophically using carbon dioxide gas as the only carbon source, yielding protein-rich biomass that can be processed further into a powder and incorporated into various food products. Since the safety of this microbial protein powder for human consumption had not been previously assessed, its genotoxic potential was evaluated employing three internationally recognized and standardized studies: a bacterial reverse mutation test, an in vitro chromosomal aberration assay in human lymphocytes, and an in vitro micronucleus test in human lymphocytes. No biologically relevant evidence of genotoxicity or mutagenicity was found.

Effects of Garlic Oil and Cinnamaldehyde on Sheep Rumen Fermentation and Microbial Populations in Rusitec Fermenters in Two Different Sampling Periods.

Garlic oil (GO) and cinnamaldehyde (CIN) have shown potential to modify rumen fermentation. The aim of this study was to assess the effects of GO and CIN on rumen fermentation, microbial protein synthesis (MPS), and microbial populations in Rusitec fermenters fed a mixed diet (50:50 forage/concentrate), as well as whether these effects were maintained over time. Six fermenters were used in two 15-day incubation runs. Within each run, two fermenters received no additive, 180 mg/L of GO, or 180 mg/L of CIN. Rumen fermentation parameters were assessed in two periods (P1 and P2), and microbial populations were studied after each of these periods. Garlic oil reduced the acetate/propionate ratio and methane production (p < 0.001) in P1 and P2 and decreased protozoal DNA concentration and the relative abundance of fungi and archaea after P1 (p < 0.05). Cinnamaldehyde increased bacterial diversity (p < 0.01) and modified the structure of bacterial communities after P1, decreased bacterial DNA concentration after P2 (p < 0.05), and increased MPS (p < 0.001). The results of this study indicate that 180 mg/L of GO and CIN promoted a more efficient rumen fermentation and increased the protein supply to the animal, respectively, although an apparent adaptive response of microbial populations to GO was observed.

Evaluating the dose-dependent effects of curcumin nano-micelles on rumen fermentation, nitrogen metabolism, and nutrient digestibility in heat-stressed fattening lambs: Implications for climate change and sustainable animal production.

Global warming threatens livestock production, especially in hot climates. This study evaluated the dose-dependent impacts of dietary curcumin nano-micelles (CNM) on rumen fermentation, nitrogen metabolism, and nutrient digestibility in heat-stressed fattening lambs. Thirty-two crossbred male lambs [ˆIle-de-France × (Dalagh × Romanov)] were utilized within the current study. The initial weight of lambs was documented as averaged by 31.2 ± 1.55 kg, while they were in their 4th to 5th months of age. Animals were fed increasing doses of dietary CNM (0, 20, 40, and 80 mg/day) over 97 days, under severe heat stress conditions with average temperature-humidity index (THI) of 24.5. Quadratic improvements (p < 0.01) occurred in weight gain, average daily gain (ADG), and feed conversion ratio (FCR) by 28.7%, 27.4%, and 23.9%, respectively, in the T40 group compared to the control. Additionally, T40 increased fiber digestion by 2.8% (p < 0.05). Furthermore, T40 quadratically improved parameters of rumen fermentation, including concentrations of NH3-N (p < 0.05), total volatile fatty acids (TVFA; p < 0.01), acetate (p < 0.05), and iso-valerate (p < 0.05), by 13.9%, 12.5%, 15.0%, and 43.5%, respectively, compared to the control. Quadratic increases were also observed in nitrogen balance (p < 0.05) and microbial protein synthesis (p < 0.01) by 19.8% and 37.6%, respectively, in the T40 group. Quadratic models estimated optimal CNM levels between 41.5 and 48.6 mg/day for multiple parameters. These findings indicate CNM at dose level of 40 mg/day could benefit heat-stressed lambs through enhanced rumen function and microbiota. Further research should refine ideal dosing for various species and production phases as higher levels adversely impacted fiber digestibility. Overall, CNM shows promise as a sustainable nutritional intervention for livestock production facing global warming.

Current trends and possibilities of typical microbial protein production approaches: a review.

Global population growth and demographic restructuring are driving the food and agriculture sectors to provide greater quantities and varieties of food, of which protein resources are particularly important. Traditional animal-source proteins are becoming increasingly difficult to meet the demand of the current consumer market, and the search for alternative protein sources is urgent. Microbial proteins are biomass obtained from nonpathogenic single-celled organisms, such as bacteria, fungi, and microalgae. They contain large amounts of proteins and essential amino acids as well as a variety of other nutritive substances, which are considered to be promising sustainable alternatives to traditional proteins. In this review, typical approaches to microbial protein synthesis processes were highlighted and the characteristics and applications of different types of microbial proteins were described. Bacteria, fungi, and microalgae can be individually or co-cultured to obtain protein-rich biomass using starch-based raw materials, organic wastes, and one-carbon compounds as fermentation substrates. Microbial proteins have been gradually used in practical applications as foods, nutritional supplements, flavor modifiers, and animal feeds. However, further development and application of microbial proteins require more advanced biotechnological support, screening of good strains, and safety considerations. This review contributes to accelerating the practical application of microbial proteins as a promising alternative protein resource and provides a sustainable solution to the food crisis facing the world.

Impact of the environmental parameters on single cell protein production and composition by Cupriavidus necator.

Due to the rapid increase in the world's population, many developing countries are facing malnutrition problems, including famine and food insecurity. Particularly, the deficiency of protein sources becomes a serious problem for human and animal nutrition. In this context, Single Cell Proteins, could be exploited as an alternative source of unconventional proteins. The aim of the study was to investigate SCP production and composition by Cupriavidus necator under various environmental conditions, temperature and pH values. A mono-factorial approach was implemented using batch bioreactor cultures under well-controlled conditions. Results were compared in terms of bacterial growth and SCP composition (proteins, nucleic acids, amino acids and elemental formula). Complementary analyses were performed by flow cytometry to study cell morphology, membrane permeability and the presence of Poly(3-hydroxybutyrate) (PHB) production. Our data confirmed the ability of C. necator to produce high amount of proteins (69 %DW at 30 °C and pH7). The results showed that temperature and pH independently impact SCP production and composition. This impact was particularly observed at the highest temperature (40 °C) and also the lowest pH value (pH5) providing lower growth rates, cell elongation, changes in granularity and lower amounts of proteins (down to 44 %DW at pH5) and nucleic acids. These low percentages were related to the production of PHB production (up to 44 %DW at 40 °C) which is the first report of a PHB accumulation in C. necator under nutrient unlimited conditions.

Evaluation of cell disruption methods for protein and coenzyme Q10 quantification in purple non-sulfur bacteria.

A recent focus has been on the recovery of single-cell protein and other nutritionally valuable bioproducts, such as Coenzyme Q10 (CoQ10) from purple non-sulfur bacteria (PNSB) biomass following wastewater treatment. However, due to PNSB's peculiar cell envelope (e.g., increased membrane cross-section for energy transduction) and relatively smaller cell size compared to well-studied microbial protein sources like yeast and microalgae, the effectiveness of common cell disruption methods for protein quantification from PNSB may differ. Thus, this study examines the efficiency of selected chemical (NaOH and EDTA), mechanical (homogenization and bead milling), physical (thermal and bath/probe sonication), and combined chemical-mechanical/physical treatment techniques on the PNSB cell lysis. PNSB biomass was recovered from the treatment of gas-to-liquid process water. Biomass protein and CoQ10 contents were quantified based on extraction efficiency. Considering single-treatment techniques, bead milling resulted in the best protein yields (p < 0.001), with the other techniques resulting in poor yields. However, the NaOH-assisted sonication (combined chemical/physical treatment technique) resulted in similar protein recovery (p = 1.00) with bead milling, with the former having a better amino acid profile. For example, close to 50% of the amino acids, such as sensitive ones like tryptophan, threonine, cystine, and methionine, were detected in higher concentrations in NaOH-assisted sonication (>10% relative difference) compared to bead-milling due to its less disruptive nature and improved solubility of amino acids in alkaline conditions. Overall, PNSB required more intensive protein extraction techniques than were reported to be effective on other single-cell organisms. NaOH was the preferred chemical for chemical-aided mechanical/physical extraction as EDTA was observed to interfere with the Lowry protein kit, resulting in significantly lower concentrations. However, EDTA was the preferred chemical agent for CoQ10 extraction and quantification. CoQ10 extraction efficiency was also suspected to be adversely influenced by pH and temperature.

Anti-Microbial Activities of Mussel-Derived Recombinant Proteins against Gram-Negative Bacteria.

Many anti-microbial peptides (AMPs) and pro-apoptotic peptides are considered as novel anti-microbial agents, distinguished by their different characteristics. Nevertheless, AMPs exhibit certain limitations, including poor stability and potential toxicity, which hinder their suitability for applications in pharmaceutics and medical devices. In this study, we used recombinant mussel adhesive protein (MAP) as a robust scaffold to overcome these limitations associated with AMPs. Mussel adhesive protein fused with functional peptides (MAP-FPs) was used to evaluate anti-microbial activities, minimal inhibitory concentration (MIC), and time-kill kinetics (TKK) assays against six of bacteria strains. MAP and MAP-FPs were proved to have an anti-microbial effect with MIC of 4 or 8 µM against only Gram-negative bacteria strains. All tested MAP-FPs killed four different Gram-negative bacteria strains within 180 min. Especially, MAP-FP-2 and -5 killed three Gram-negative bacteria strain, including E. coli, S. typhimurium, and K. pneumoniae, within 10 min. A cytotoxicity study using Vero and HEK293T cells indicated the safety of MAP and MAP-FP-2 and -3. Thermal stability of MAP-FP-2 was also validated by HPLC analysis at an accelerated condition for 4 weeks. This study identified that MAP-FPs have novel anti-microbial activity, inhibiting the growth and rapidly killing Gram-negative bacteria strains with high thermal stability and safety.

A comprehensive report on valorization of waste to single cell protein: strategies, challenges, and future prospects.

The food insecurity due to a vertical increase in the global population urgently demands substantial advancements in the agricultural sector and to identify sustainable affordable sources of nutrition, particularly proteins. Single-cell protein (SCP) has been revealed as the dried biomass of microorganisms such as algae, yeast, and bacteria cultivated in a controlled environment. Production of SCP is a promising alternative to conventional protein sources like soy and meat, due to quicker production, minimal land requirement, and flexibility to various climatic conditions. In addition to protein production, it also contributes to waste management by converting it into food and feed for both human and animal consumption. This article provides an overview of SCP production, including its benefits, safety, acceptability, and cost, as well as limitations that constrains its maximum use. Furthermore, this review criticizes the downstream processing of SCP, encompassing cell wall disruption, removal of nucleic acid, harvesting of biomass, drying, packaging, storage, and transportation. The potential applications of SCP, such as in food and feed as well as in the production of bioplastics, emulsifiers, and as flavoring agents for baked food, soup, and salad, are also discussed.

Enhanced Microbial Protein Production from CO2 and Air by a MoS2 Catalyzed Bioelectrochemical System.

Carbon dioxide can be relatively easily reduced to organic matter in a bioelectrochemical system (BES). However, due to insufficient reduction force from in-situ hydrogen evolution, it is difficult for nitrogen reduction. In this study, MoS2 was firstly used as an electrocatalyst for the simultaneous reduction of CO2 and N2 to produce microbial protein (MP) in a BES. Cell dry weight (CDW) could reach 0.81±0.04 g/L after 14 d operation at -0.7 V (vs. RHE), which was 108±3 % higher than that from non-catalyst control group (0.39±0.01 g/L). The produced protein had a better amino acid profile in the BES than that in a direct hydrogen system (DHS), particularly for proline (Pro). Besides, MoS2 promoted the growth of bacterial cell on an electrode and improved the biofilm extracellular electron transfer (EET) by microscopic observation and electrochemical characterization of MoS2 biocathode. The composition of the microbial community and the relative abundance of functional enzymes revealed that MoS2 as an electrocatalyst was beneficial for enriching Xanthobacter and enhancing CO2 and N2 reduction by electrical energy. These results demonstrated that an efficient strategy to improve MP production of BES is to use MoS2 as an electrocatalyst to shift amino acid profile and microbial community.

Effects of nitrogen supply on hydrogen-oxidizing bacterial enrichment to produce microbial protein: Comparing nitrogen fixation and ammonium assimilation.

To investigate the effects of nitrogen source supply on microbial protein (MP) production by hydrogen-oxidizing bacteria (HOB) under continuous feed gas provision, a sequencing batch culture comparison (N2 fixation versus ammonium assimilation) was performed. The results confirmed that even under basic cultivation conditions, N2-fixing HOB (NF-HOB) communities showed higher levels of CO2 and N2 fixation (190.45 mg/L Δ CODt and 11.75 mg/L Δ TNbiomass) than previously known, with the highest biomass yield being 0.153 g CDW/g COD-H2. Rich ammonium stimulated MP synthesis and the biomass accumulation of communities (increased by 7.4 ～ 14.3 times), presumably through the enhancement of H2 and CO2 absorption. The micro mechanism may involve encouraging the enrichment of species like Xanthobacter and Acinetobacter then raising the abundance of nitrogenase and glutamate synthase to facilitate the nitrogen assimilation. This would provide NF-HOB with ideas for optimizing their MP synthesis activity.

Dietary replacement of soybean meal with heat-treated grain soybean in diets of feedlot-finished beef cattle: impacts on intake, digestibility, and ruminal parameters.

The objective of this study was to assess the impact of increasing levels of heat-treated soybean in the diet of crossbred cattle during the finishing phase on nutrient intake and digestibility, ruminal parameters, digesta passage rate, nitrogen balance, and microbial protein synthesis. Five steers, crossbred 7/8 Jersey x Zebu, fitted with rumen cannulas and with an average weight of 350 ± 50 kg, were utilized. The experimental treatments consisted of 0, 7, 14, 21, and 28% inclusion of heat-treated soybean in the total diet dry matter. The animals were randomly allocated in a 5 × 5 Latin square design. Evaluation of the animals took place over five experimental periods, each lasting 20 days. During each experimental period, the first 15 days were allocated for animal adaptation to the experimental diets, followed by five days of data collection. No significant differences were observed among the diets in terms of dry matter intake (average of 6.57 kg day-1; P = 0.615) and organic matter intake (average of 6.23 kg day-1; P = 0.832). However, heat-treated soybean had a significant impact on the digestibility of dry matter (P = 0.02), organic matter (P = 0.01), crude protein (P < 0.01), and neutral detergent fiber (P < 0.01). There was no observed change on microbial protein synthesis (average of 409.6 g day-1) in animals with the inclusion of heat-treated soybean in the diets. With each 1% inclusion of heat-treated soybean in the cattle diet, there was an increase of 0.00754 units in ruminal pH values and a reduction of 0.75839 mg dL-1 in ruminal ammoniacal nitrogen values. This study suggests that heat-treated soybean can be included in up to 15% of the dry matter in diets for finishing feedlot cattle.

Estimating Microbial Protein Synthesis in the Rumen-Can 'Omics' Methods Provide New Insights into a Long-Standing Question?

Rumen microbial protein synthesis (MPS) provides at least half of the amino acids for the synthesis of milk and meat protein in ruminants. As such, it is fundamental to global food protein security. Estimating microbial protein is central to diet formulation, maximising nitrogen (N)-use efficiency and reducing N losses to the environment. Whilst factors influencing MPS are well established in vitro, techniques for in vivo estimates, including older techniques with cannulated animals and the more recent technique based on urinary purine derivative (UPD) excretion, are subject to large experimental errors. Consequently, models of MPS used in protein rationing are imprecise, resulting in wasted feed protein and unnecessary N losses to the environment. Newer 'omics' techniques are used to characterise microbial communities, their genes and resultant proteins and metabolites. An analysis of microbial communities and genes has recently been used successfully to model complex rumen-related traits, including feed conversion efficiency and methane emissions. Since microbial proteins are more directly related to microbial genes, we expect a strong relationship between rumen metataxonomics/metagenomics and MPS. The main aims of this review are to gauge the understanding of factors affecting MPS, including the use of the UPD technique, and explore whether omics-focused studies could improve the predictability of MPS, with a focus on beef cattle.