Biochar addition reduces non-CO2 greenhouse gas emissions during composting of human excreta and cattle manure.

Ecological sanitation combined with thermophilic composting is a viable option to transform human excreta into a stabilized, pathogen-free, and nutrient-rich fertilizer. In combination with suitable bulking materials such as sawdust and straw, and additives such as biochar, this could also be a suitable waste management strategy for reducing greenhouse gas (GHG) emissions. In this study, we conducted a 143-days thermophilic composting of human excreta or cattle manure together with teff straw, organic waste, and biochar to investigate the effect that biochar has on GHG (CO2 , N2 O, and CH4 ) and NH3 emissions. The composting was performed in wooden boxes (1.5 × 1.5 × 1.4 m3 ), GHG were measured by using a portable FTIR gas analyzer and NH3 was sampled as ammonium in an H2 SO4 trap. We found that the addition of biochar significantly reduced CH4 emissions by 91% in the cattle manure compost, and N2 O emissions by 56%-57% in both humanure and cattle manure composts. Overall, non-CO2 GHG emissions were reduced by 51%-71%. In contrast, we did not observe a significant biochar effect on CO2 and NH3 emissions. Previous data already showed that it is possible to sanitize human fecal material when using this composting method. Our results suggest that thermophilic composting with biochar addition is a safe and cost-effective waste management practice for producing a nutrient-rich fertilizer from human excreta, while reducing GHG emissions at the same time.

Fate of Horizontal-Gene-Transfer Markers and Beta-Lactamase Genes during Thermophilic Composting of Human Excreta.

Thermophilic composting is a suitable treatment for the recycling of organic wastes for agriculture. However, using human excreta as feedstock for composting raises concerns about antibiotic resistances. We analyzed samples from the start and end of a thermophilic composting trial of human excreta, together with green cuttings and straw, with and without biochar. Beta-lactamase genes blaCTX-M, blaIMP, and blaTEM conferring resistance to broad-spectrum beta-lactam antibiotics, as well as horizontal gene transfer marker genes, intI1 and korB, were quantified using qPCR. We found low concentrations of the beta-lactamase genes in all samples, with non-significant mean decreases in blaCTX-M and blaTEM copy numbers and a mean increase in blaIMP copy numbers. The decrease in both intI1 and korB genes from start to end of composting indicated that thermophilic composting can decrease the horizontal spread of resistance genes. Thus, thermophilic composting can be a suitable treatment for the recycling of human excreta.

Metagenomic Insights Into the Changes of Antibiotic Resistance and Pathogenicity Factor Pools Upon Thermophilic Composting of Human Excreta.

In times of climate change, practicing a form of sustainable, climate-resilient and productive agriculture is of primordial importance. Compost could be one form of sustainable fertilizer, which is increasing humus, water holding capacity, and nutrient contents of soils. It could thereby strengthen agriculture toward the adverse effects of climate change, especially when additionally combined with biochar. To get access to sufficient amounts of suitable materials for composting, resources, which are currently treated as waste, such as human excreta, could be a promising option. However, the safety of the produced compost regarding human pathogens, pharmaceuticals (like antibiotics) and related resistance genes must be considered. In this context, we have investigated the effect of 140- and 154-days of thermophilic composting on the hygienization of human excreta and saw dust from dry toilets together with straw and green cuttings with and without addition of biochar. Compost samples were taken at the beginning and end of the composting process and metagenomic analysis was conducted to assess the fate of antibiotic resistance genes (ARGs) and pathogenicity factors of the microbial community over composting. Potential ARGs conferring resistance to major classes of antibiotics, such as beta-lactam antibiotics, vancomycin, the MLSB group, aminoglycosides, tetracyclines and quinolones were detected in all samples. However, relative abundance of ARGs decreased from the beginning to the end of composting. This trend was also found for genes encoding type III, type IV, and type VI secretion systems, that are involved in pathogenicity, protein effector transport into eukaryotic cells and horizontal gene transfer between bacteria, respectively. The results suggest that the occurrence of potentially pathogenic microorganisms harboring ARGs declines during thermophilic composting. Nevertheless, ARG levels did not decline below the detection limit of quantitative PCR (qPCR). Thresholds for the usage of compost regarding acceptable resistance gene levels are yet to be evaluated and defined.

Thermophilic Composting of Human Feces: Development of Bacterial Community Composition and Antimicrobial Resistance Gene Pool.

In times of climate change, practicing sustainable, climate-resilient, and productive agriculture is of primordial importance. Compost from different resources, now treated as wastes, could be one form of sustainable fertilizer creating a resilience of agriculture to the adverse effects of climate change. However, the safety of the produced compost regarding human pathogens, pharmaceuticals, and related resistance genes must be considered. We have assessed the effect of thermophilic composting of dry toilet contents, green cuttings, and straw, with and without biochar, on fecal indicators, the bacterial community, and antibiotic resistance genes (ARGs). Mature compost samples were analyzed regarding fecal indicator organisms, revealing low levels of Escherichia coli that are in line with German regulations for fertilizers. However, one finding of Salmonella spp. exceeded the threshold value. Cultivation of bacteria from the mature compost resulted in 200 isolates with 36.5% of biosafety level 2 (BSL-2) species. The majority is known as opportunistic pathogens that likewise occur in different environments. A quarter of the isolated BSL-2 strains exhibited multiresistance to different classes of antibiotics. Molecular analysis of total DNA before and after composting revealed changes in bacterial community composition and ARGs. 16S rRNA gene amplicon sequencing showed a decline of the two most abundant phyla Proteobacteria (start: 36-48%, end: 27-30%) and Firmicutes (start: 13-33%, end: 12-16%), whereas the abundance of Chloroflexi, Gemmatimonadetes, and Planctomycetes rose. Groups containing many human pathogens decreased during composting, like Pseudomonadales, Bacilli with Bacillus spp., or Staphylococcaceae and Enterococcaceae. Gene-specific PCR showed a decline in the number of detectable ARGs from 15 before to 8 after composting. The results reveal the importance of sufficiently high temperatures lasting for a sufficiently long period during the thermophilic phase of composting for reducing Salmonella to levels matching the criteria for fertilizers. However, most severe human pathogens that were targeted by isolation conditions were not detected. Cultivation-independent analyses also indicated a decline in bacterial orders comprising many pathogenic bacteria, as well as a decrease in ARGs. In summary, thermophilic composting could be a promising approach for producing hygienically safe organic fertilizer from ecological sanitation.

Nutrient dynamics during composting of human excreta, cattle manure, and organic waste affected by biochar.

Ecological sanitation via thermophilic composting could be a promising solution to the lack of sanitation and limited access to fertilizers, particularly in developing countries. Here, we conducted a 185-d thermophilic composting experiment with human excreta, and separately with cattle manure, mixed with kitchen scraps, teff [Eragrostis tef (Zuccagni) Trotter] straw, sawdust, and biochar (BC) by using an appropriate-technology approach. We followed the dynamics of the most important macronutrients (N, P, K), temperature, moisture, pH, electrical conductivity, cation exchange capacity, as well as content of organic matter, organic C, Ca, Mg, and micronutrients throughout the process. Low N (<47%), P (<9%), K (<11%), Ca (<18%), and Mg (<21%) losses and the temperature profile indicated a well-functioning thermophilic composting process. Compost temperature was >60 °C for 7, 6, 5, and 8 consecutive days for treatments containing human excreta, human excreta amended with BC, cattle manure, and cattle manure amended with BC, respectively, suggesting a final compost product free of pathogens. The compost mixture with cattle manure and BC reached a significantly higher temperature than the same variant without BC, with a maximum value of 65.9 °C on Day 6. For all treatments, final germination index values >100% indicated compost maturity and the absence of phytotoxic substances. Biochar addition reduced losses of organic matter (18-23%), C (33-42%), and N (49-100%) and decreased the amount of extractable NO3 - (32-36%) in the final compost. The tested ecological sanitation concept via thermophilic composting is thus a promising strategy to improve access to cheap fertilizer by safe and sustainable sanitation and waste management.

Steroid Biomarkers Revisited - Improved Source Identification of Faecal Remains in Archaeological Soil Material.

Steroids are used as faecal markers in environmental and in archaeological studies, because they provide insights into ancient agricultural practices and the former presence of animals. Up to now, steroid analyses could only identify and distinguish between herbivore, pig, and human faecal matter and their residues in soils and sediments. We hypothesized that a finer differentiation between faeces of different livestock animals could be achieved when the analyses of several steroids is combined (Δ5-sterols, 5α-stanols, 5ß-stanols, epi-5ß-stanols, stanones, and bile acids). We therefore reviewed the existing literature on various faecal steroids from livestock and humans and analysed faeces from old livestock breed (cattle, horse, donkey, sheep, goat, goose, and pig) and humans. Additionally, we performed steroid analyses on soil material of four different archaeological periods (sites located in the Lower Rhine Basin, Western Germany, dating to the Linearbandkeramik, Urnfield Period / Bronze Age, Iron Age, Roman Age) with known or supposed faecal inputs. By means of already established and newly applied steroid ratios of the analysed faeces together with results from the literature, all considered livestock faeces, except sheep and cattle, could be distinguished on the basis of their steroid signatures. Most remarkably was the identification of horse faeces (via the ratio: epi-5ß-stigmastanol: 5ß-stigmastanol + epicoprostanol: coprostanol; together with the presence of chenodeoxycholic acid) and a successful differentiation between goat (with chenodeoxycholic acid) and sheep/cattle faeces (without chenodeoxycholic acid). The steroid analysis of archaeological soil material confirmed the supposed faecal inputs, even if these inputs had occurred several thousand years ago.

Organic fertilization and sufficient nutrient status in prehistoric agriculture?--Indications from multi-proxy analyses of archaeological topsoil relicts.

Neolithic and Bronze Age topsoil relicts revealed enhanced extractable phosphorus (P) and plant available inorganic P fractions, thus raising the question whether there was targeted soil amelioration in prehistoric times. This study aimed (i) at assessing the overall nutrient status and the soil organic matter content of these arable topsoil relicts, and (ii) at tracing ancient soil fertilizing practices by respective stable isotope and biomarker analyses. Prehistoric arable topsoils were preserved in archaeological pit fillings, whereas adjacent subsoils served as controls. One Early Weichselian humic zone represented the soil status before the introduction of agriculture. Recent topsoils served as an additional reference. The applied multi-proxy approach comprised total P and micronutrient contents, stable N isotope ratios, amino acid, steroid, and black carbon analyses as well as soil color measurements. Total contents of P and selected micronutrients (I, Cu, Mn, Mo, Se, Zn) of the arable soil relicts were above the limits for which nutrient deficiencies could be assumed. All pit fillings exhibited elevated δ15N values close to those of recent topsoils (δ15N>6 to 7‰), giving first hints for prehistoric organic N-input. Ancient legume cultivation as a potential source for N input could not be verified by means of amino acid analysis. In contrast, bile acids as markers for faecal input exhibited larger concentrations in the pit fillings compared with the reference and control soils indicating faeces (i.e. manure) input to Neolithic arable topsoils. Also black carbon contents were elevated, amounting up to 38% of soil organic carbon, therewith explaining the dark soil color in the pit fillings and pointing to inputs of burned biomass. The combination of different geochemical analyses revealed a sufficient nutrient status of prehistoric arable soils, as well as signs of amelioration (inputs of organic material like charcoal and faeces-containing manure).

Greenhouse gas production in mixtures of soil with composted and noncomposted biochars is governed by char-associated organic compounds.

Biochar application to soil has the potential to increase soil productivity while reducing anthropogenic greenhouse gas (GHG) emissions to the atmosphere. However, techniques for conditioning this material for maximizing its effects as a soil amendment require elucidation. We examined changes of organic matter associated with two biochars after 175 d of composting and the resulting effects on GHG emissions during a 150-d incubation period. Composting decreased the amount of organic compounds that could be thermally released from the biochars and affected their molecular nature. These thermally desorbable organic compounds from initial biochars likely stimulated the oxidation of CH and inhibited the production of NO in soil-biochar mixtures. However, these reductions of GHG emissions disappeared together with thermally desorbable organic compounds after the composting of chars. Instead, addition of composted gasification coke and charcoal stimulated the formation of CH and increased NO emissions by 45 to 56%. Nitrous oxide emissions equaled 20% of the total amount of N added with composted biochars, suggesting that organic compounds and N sorbed by the chars during composting fueled GHG production. The transient nature of the suppression of CH and NO production challenges the long-term GHG mitigation potential of biochar in soil.

Biochar affected by composting with farmyard manure.

Biochar applications to soils can improve soil fertility by increasing the soil's cation exchange capacity (CEC) and nutrient retention. Because biochar amendment may occur with the applications of organic fertilizers, we tested to which extent composting with farmyard manure increases CEC and nutrient content of charcoal and gasification coke. Both types of biochar absorbed leachate generated during the composting process. As a result, the moisture content of gasification coke increased from 0.02 to 0.94 g g, and that of charcoal increased from 0.03 to 0.52 g g. With the leachate, the chars absorbed organic matter and nutrients, increasing contents of water-extractable organic carbon (gasification coke: from 0.09 to 7.00 g kg; charcoal: from 0.03 to 3.52 g kg), total soluble nitrogen (gasification coke: from not detected to 705.5 mg kg; charcoal: from 3.2 to 377.2 mg kg), plant-available phosphorus (gasification coke: from 351 to 635 mg kg; charcoal: from 44 to 190 mg kg), and plant-available potassium (gasification coke: from 6.0 to 15.3 g kg; charcoal: from 0.6 to 8.5 g kg). The potential CEC increased from 22.4 to 88.6 mmol kg for the gasification coke and from 20.8 to 39.0 mmol kg for the charcoal. There were little if any changes in the contents and patterns of benzene polycarboxylic acids of the biochars, suggesting that degradation of black carbon during the composting process was negligible. The surface area of the biochars declined during the composting process due to the clogging of micropores by sorbed compost-derived materials. Interactions with composting substrate thus enhance the nutrient loads but alter the surface properties of biochars.