Activity- and Enrichment-Based Metaproteomics Insights into Active Urease from the Rumen Microbiota of Cattle

Regulation of microbial urease activity plays a crucial role in improving the utilization efficiency of urea and reducing nitrogen emissions to the environment for ruminant animals. Dealing with the diversity of microbial urease and identifying highly active urease as the target is the key for future regulation. However, the identification of active urease in the rumen is currently limited due to large numbers of uncultured microorganisms. In the present study, we describe an activity- and enrichment-based metaproteomic analysis as an approach for the discovery of highly active urease from the rumen microbiota of cattle. We conducted an optimization method of protein extraction and purification to obtain higher urease activity protein. Cryomilling was the best choice among the six applied protein extraction methods (ultrasonication, bead beating, cryomilling, high-pressure press, freeze-thawing, and protein extraction kit) for obtaining protein with high urease activity. The extracted protein by cryomilling was further enriched through gel filtration chromatography to obtain the fraction with the highest urease activity. Then, by using SDS-PAGE, the gel band including urease was excised and analyzed using LC-MS/MS, searching against a metagenome-derived protein database. Finally, we identified six microbial active ureases from 2225 rumen proteins, and the identified ureases were homologous to those of Fibrobacter and Treponema. Moreover, by comparing the 3D protein structures of the identified ureases and known ureases, we found that the residues in the β-turn of flap regions were nonconserved, which might be crucial in influencing the flexibility of flap regions and urease activity. In conclusion, the active urease from rumen microbes was identified by the approach of activity- and enrichment-based metaproteomics, which provides the target for designing a novel efficient urease inhibitor to regulate rumen microbial urease activity.

Quantification and comparison of ecosystem services of grasslands versus another fodder crop (maize) based on mineral nitrogen content in the 60–90 cm soil layer

Grasslands provide many ecosystem services. Apart from being a source of fodder for animals, grasslands regulate water and soil quality by reducing nitrogen emissions to the environment. The aim of the study was to determine the biophysical and monetary value of ecosystem services of grassland based on the “mineral nitrogen content in the soil layer 60–90 cm” indicator depending on the method of use and the type of soil, against the cultivation of maize for green fodder. The study area encompassed three provinces, different in terms of soil use, livestock population and intensity of grassland use. The investigation indicated that the value of ecosystem services provided by grasslands varied spatially and depended on the type of use and type of soil. In mineral soils, the lowest levels of this index were recorded from sites used for pasturing, while the highest levels were found under maize crops. In organic soils (without maize crops), the smallest losses of Nmin were observed in meadows while the highest losses were in pastures. Nmin losses in organic soils were higher than in mineral soils. The losses observed were highest in Opolskie Province, followed by Podlaskie Province, with the lowest losses in Lubelskie Province.

Diet optimization for dairy cows to reduce ammonia emissions

Feeding livestock a balanced diet with a differentiated crude protein (CP) content, depending on the lactation phase can reduce nitrogen emissions from livestock excrement and urine. A higher content of non-starch polysaccharides in livestock diets improves feed absorption in the livestock body and, consequently, nitrogen is emitted more from protein present in livestock manure than from urea acid present in livestock urine. The aim of the study is to calculate the ammonia emission reduction potential in Latvia by optimizing the feeding of dairy cows and ensuring life longevity, as well as provide justification for ammonia emission reduction in dairy farms. Calculations made by using the NorFor Model for optimization of dairy cow (Bos primigenius f. taurus) diets revealed that compared with lowyielding cows, a higher CP content diet fed to high-yielding cows at the beginning of lactation increased the amount of nitrogen (N) in their excrement and urine by 90–180 g d-1. Reducing the CP content in the cow diet by an average of 10 g kg-1 dry matter (DM) during mid-lactation resulted in the same trend. Reducing the CP content in the cow diet during late lactation and the dry period by another 20–30 g kg-1 of DM, N emissions from excrement and urine significantly decreased. Increasing the lifespan of dairy cows also means reducing ammonia emissions from the farm. By increasing the number of lactations per cow on dairy farm, it is possible to reduce the number of heifers per cow. The total reduction of ammonia emissions in Latvia was calculated based on a long-term projection of a decrease of 0.1 heifer per dairy cow. Ammonia emissions could be reduced by 0.051 kt by decreasing the number of heifers by 12.54 thou. at the planned increase in the lifespan of dairy cows by 2030.

Factors Influencing Proteolysis and Protein Utilization in the Intestine of Pigs: A Review

Pigs are among the most important farm animals for meat production worldwide. In order to meet the amino acid requirements of the animals, pigs rely on the regular intake of proteins and amino acids with their feed. Unfortunately, pigs excrete about two thirds of the used protein, and production of pork is currently associated with a high emission of nitrogen compounds resulting in negative impacts on the environment. Thus, improving protein efficiency in pigs is a central aim to decrease the usage of protein carriers in feed and to lower nitrogen emissions. This is necessary as the supply of plant protein sources is limited by the yield and the cultivable acreage for protein plants. Strategies to increase protein efficiency that go beyond the known feeding options have to be investigated considering the characteristics of the individual animals. This requires a deep understanding of the intestinal processes including enzymatic activities, capacities of amino acid transporters and the microbiome. This review provides an overview of these physiological factors and the respective analyses methods.

Drivers of global nitrogen emissions

Nitrogen is crucial for sustaining life. However, excessive reactive nitrogen (Nr) in the form of ammonia, nitrates, nitrogen oxides or nitrous oxides affects the quality of water, air and soil, resulting in human health risks. This study aims to assess the drivers of Nr emissions by analysing six determinants: nitrogen efficiency (Nr emissions per unit of production), production recipe (inter-sectoral dependencies), final demand composition (consumption baskets of households), final demand destination (consumption vs. investment balance), affluence (final consumption per capita) and population. To this end, we construct a detailed multi-regional input-output database featuring data on international trade between 186 countries to undertake a global structural decomposition analysis of a change in global Nr emissions from 1997-2017. Our analysis shows that nitrogen efficiency has improved over the assessed time-period, however affluence, final demand destination and population growth have resulted in an overall increase in Nr emissions. We provide a global perspective of the drivers of nitrogen emissions at a detailed country level, and breakdown the change in emissions into contribution from domestic footprint and rest-of-world footprint. We highlight that food production coupled with growing international trade is increasing Nr emissions worldwide.

Urban Metabolism of Food-Sourced Nitrogen among Different Income Households: A Case Study Based on Large Sample Survey in Xiamen City, China

Food consumption is fundamental for urban households if they are to sustain production and daily life. Nitrogen resulting from food consumption has significantly contributed to pollutant emissions in urban ecosystems. Taking Xiamen city, a rapid urbanizing area of southeast China as a case study, we evaluated the food-sourced nitrogen consumption of households based on a large simple onsite questionnaire survey, as well as differences between households in the consumption of plant-based and animal-based foods. A material flow analysis (MFA) was conducted to simulate the urban metabolism of food-sourced nitrogen and environmental emissions among different income groups. The impacts of household attributes, plant-based food consumption, and animal-based food consumption on environmental nitrogen emissions were examined with a structural equation model (SEM). Our results show that the surveyed households’ diets were more plant-based and less animal-based. Aquatic products and livestock were the source of 43.7% of food-sourced nitrogen, and 84.5% of the food-sourced nitrogen was discharge into the environment through direct discharge and waste treatment. Soil, water, and air emissions accounted for 62.8%, 30.1%, and 7.1% of the food-sourced nitrogen, respectively. Household income, household size, and household area are all associated with accelerating increases of nitrogen emissions released into the environment, though middle-income group households have the highest food-sourced environmental nitrogen emissions. On this basis, we discuss how to better manage the urban metabolism of food-sourced nitrogen, so as to improve urban household consumption, lower nitrogen emissions, and improve food security.

Interannual variation of reactive nitrogen emissions and their impacts on PM2.5 air pollution in China during 2005-2015

Emissions of reactive nitrogen as ammonia (NH3) and nitrogen oxides (NOx), together with sulfur dioxide (SO2), contribute to formation of secondary PM2.5 in the atmosphere. Satellite observations of atmospheric NH3, NO2, and SO2 levels since the 2000s provide valuable information to constrain the spatial and temporal variability of their emissions. Here we present a bottom-up Chinese NH3 emission inventory combined with top-down estimates of Chinese NOx and SO2 emissions using OMI satellite observations, aiming to quantify the interannual variations of reactive nitrogen emissions in China and their contributions to PM2.5 air pollution over 2005–2015. We find small interannual changes in the total Chinese anthropogenic NH3 emissions during 2005–2016 (12.0–13.3 Tg with over 85% from agricultural sources), but large interannual change in top-down Chinese NOx and SO2 emissions. Chinese NOx emissions peaked around 2011 and declined by 22% during 2011–2015, and Chinese SO2 emissions declined by 55% in 2015 relative to that in 2007. Using the GEOS-Chem chemical transport model simulations, we find that rising atmospheric NH3 levels in eastern China since 2011 as observed by IASI and AIRS satellites are mainly driven by rapid reductions in SO2 emissions. The 2011–2015 Chinese NOx emission reductions have decreased regional annual mean PM2.5 by 2.3–3.8 μg m-3. Interannual PM2.5 changes due to NH3 emission changes are relatively small, but further control of agricultural NH3 emissions can be effective for PM2.5 pollution mitigation in eastern China.