A newly identified IncY plasmid from multi-drug-resistant Escherichia coli isolated from dairy cattle feces in Poland.

Comprehensive whole-genome sequencing was performed on two multi-drug-resistant Escherichia coli strains isolated from cattle manure from a typical dairy farm in Poland in 2020. The identified strains are resistant to beta-lactams, aminoglycosides, tetracyclines, trimethoprim/sulfamethoxazole, and fluoroquinolones. The complete sequences of the harbored plasmids revealed antibiotic-resistance genes located within many mobile genetic elements (e.g., insertional sequences or transposons) and genes facilitating conjugal transfer or promoting horizontal gene transfer. These plasmids are hitherto undescribed. Similar plasmids have been identified, but not in Poland. The identified plasmids carried resistance genes, including the tetracycline resistance gene tet(A), aph family aminoglycoside resistance genes aph(3″)-lb and aph (6)-ld, beta-lactam resistance genes blaTEM-1 and blaCTX-M-15, sulfonamide resistance gene sul2, fluoroquinolone resistance gene qnrS1, and the trimethoprim resistance gene dfrA14. The characterized resistance plasmids were categorized into the IncY incompatibility group, indicating a high possibility for dissemination among the Enterobacteriaceae. While similar plasmids (99% identity) have been found in environmental and clinical samples, none have been identified in farm animals. These findings are significant within the One Health framework, as they underline the potential for antimicrobial-resistant E. coli from livestock and food sources to be transmitted to humans and vice versa. It highlights the need for careful monitoring and strategies to limit the spread of antibiotic resistance in the One Health approach. IMPORTANCE: This study reveals the identification of new strains of antibiotic-resistant Escherichia coli in cattle manure from a dairy farm in Poland, offering critical insights into the spread of drug resistance. Through whole-genome sequencing, researchers discovered novel plasmids within these bacteria, which carry genes resistant to multiple antibiotics. These findings are particularly alarming, as these plasmids can transfer between different bacterial species, potentially escalating the spread of antibiotic resistance. This research underscores the vital connection between the health of humans, animals, and the environment, emphasizing the concept of One Health. It points to the critical need for global vigilance and strategies to curb the proliferation of antibiotic resistance. By showcasing the presence of these strains and their advanced resistance mechanisms, the study calls for enhanced surveillance and preventive actions in both agricultural practices and healthcare settings to address the imminent challenge of antibiotic-resistant bacteria.

Effect of Cow Dung Additions on Tropical and Mediterranean Earth Mortars-Mechanical Performance and Water Resistance.

Cow dung (CD) is a material that has been used for millennia by humanity as a stabilizer in earth building techniques in vernacular architecture. However, this stabilization has been little addressed scientifically. In this study, the effect of CD additions was assessed on earth mortars produced with one type of earth from Brazil and two other types from Portugal (from Monsaraz and Caparica). The effect of two volumetric proportions of CD additions were assessed: 10% and 20% of earth + sand. The German standard DIN 18947 was used to perform the physical and mechanical tests, and classify the mortars. In comparison to the reference mortars without CD, the additions reduced linear shrinkage and cracking. An increase in flexural and compressive strengths was not observed only in mortars produced with earth from Monsaraz. In mortars produced with the earth from Caparica, the addition of 10% of CD increased flexural strength by 15% and compressive strength by 34%. For mortars produced with the earth from Brazil, the addition of 10% of CD increased these mechanical strengths by 40%. The increase in adhesive strength and water resistance promoted by the CD additions was observed in mortars produced with all three types of earth. Applied on ceramic brick, the proportion of 10% of CD increased the adherence by 100% for the three types of earth. Applied on adobe, the same proportion of CD also increased it more than 50%. For the water immersion test, the CD additions made possible for the mortar specimens not to disintegrate after a 30 min immersion, with the 20% proportion being more efficient. The effects of the CD on mechanical performance, including adhesion, were more significant on the tropical earth mortars but the effects on water resistance were more significant on the Mediterranean earthen mortars. CD has shown its positive effects and potential for both tropical and Mediterranean earthen plasters and renders tested, justifying being further studied as an eco-efficient bio-stabilizer.

Material flow analysis of heavy metals in large-scale cattle farms and ecological risk assessment of cattle manure application to fields.

This study bridges the knowledge gap pertaining to the pathways of heavy metal accumulation and migration within the industrial chain of large-scale cattle farms. Two such farms in Shaanxi serve as a basis for our exploration into Zn, Cu, Cr, Pb, As, and Cd dynamics. Employing material flow analysis complemented by predictive models, we evaluate the potential ecological risks of arable soil from heavy metal influx via manure application. Our findings indicate that Zn and Cu predominate the heavy metal export from these operations, composing up to 60.00%-95.67% of their total content. Predictive models based on 2021 data reveal a potential increase in Cd soil concentration by 0.08 mg/kg by 2035, insinuating a reduced safe usage period for cattle manure at less than 50 years. Conversely, projections from 2022 data point towards a gradual Cu rise in soil, reaching risk threshold levels after 126 years. These outcomes inform limitations in cattle manure utilisation strategies, underscoring Cu and Cd content as key barriers. The study underscores the criticality of continuous heavy metal surveillance within farm by products to ensure environmental protection and sustainable agricultural practices.

Insight into N2O emission and denitrifier communities under different aeration intensities in composting of cattle manure from perspective of multi-factor interaction analysis.

Nitrous oxide (N2O) emission from composting is a significant contributor to greenhouse effect and ozone depletion, which poses a threat to environment. To address the challenge of mitigating N2O emission during composting, this study investigated the response of N2O emission and denitrifier communities (detected by metagenome sequencing) to aeration intensities of 6 L/min (C6), 12 L/min (C12), and 18 L/min (C18) in cattle manure composting using multi-factor interaction analysis. Results showed that N2O emission occurred mainly at mesophilic phase. Cumulative N2O emission (QN2O, 9.79 mg·kg-1 DW) and total nitrogen loss (TN loss, 16.40 %) in C12 composting treatment were significantly lower than those in the other two treatments. The lower activity of denitrifying enzymes and the more complex and balanced network of denitrifiers and environmental factors might be responsible for the lower N2O emission. Denitrification was confirmed to be the major pathway for N2O production. Moisture content (MC) and Luteimonas were the key factors affecting N2O emission, and nosZ-carrying denitrifier played a significant role in reducing N2O emission. Although relative abundance of nirS was lower than that of nirK significantly (P < 0.05), nirS was the key gene influencing N2O emission. Community composition of denitrifier varied significantly with different aeration treatments (R2 = 0.931, P = 0.001), and Achromobacter was unique to C12 at mesophilic phase. Physicochemical factors had higher effect on QN2O, whereas denitrifying genes, enzymes and NOX- had lower effect on QN2O in C12. The complex relationship between N2O emission and the related factors could be explained by multi-factor interaction analysis more comprehensively. This study provided a novel understanding of mechanism of N2O emission regulated by aeration intensity in composting.

Enhanced anaerobic co-digestion of cattle manure with food waste and pig manure: Statistical optimization of pretreatment condition and substrate mixture ratio.

This study investigated the optimal pretreatment condition and mixture ratio of cattle manure (CM) for its efficient anaerobic co-digestion (AcoD) with food waste (FW) and pig manure (PM). The pretreatment performances of thermal (TM), microwave (MW), and ultrasound (US) technologies and the AcoD performance were statistically and experimentally evaluated at various mixture ratios of CM, FW, and PM. The results revealed that the most effective pretreatment condition with the TM, MW, and US pretreatments was 129.3 °C for 49.6 min, 824.2 W for 7.3 min, and 418.0 W for 36.3 min, respectively. The best AcoD performance of optimally pretreated CM (PCM) was achieved when 30.5 % PCM was mixed with 42.5 % FW and 27.0 % PM. A long-term evaluation showed that the start-up rate for the anaerobic mono-digestion of PCM was 2.3 times faster than that of CM and the amount of methane produced was 4.7 times higher; process stability was thus preferentially maintained under a higher organic loading rate (OLR) (2.0 kg-VS/m3∙d). The start-up rate for the AcoD of PCM with FW and PM was 1.2 times higher than that of the AcoD of CM with FW and PM. Although the performance gap between the AcoD reactors after steady state was not significantly different, the PCM AcoD reactor provided a more stable operation under a higher OLR (5.0 kg-VS/m3∙d). This study demonstrates that the pretreatment and co-digestion of CM could significantly enhance the production of biogas and improve process stability.

Effects of hydrothermal treatment on the reduction of antibiotic-resistant Escherichia coli and antibiotic resistance genes and the fertilizer potential of liquid product from cattle manure.

In this study, the reduction in the abundance of antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs) and the fertilizer potential of liquid products from hydrothermally treated cattle manure were investigated. Hydrothermal treatment (HTT) was conducted under different reaction temperatures (125, 150, 175 and 200 °C) and retention times (60, 90 and 120 min). The total organic carbon (TOC) and total nitrogen (TN) of the liquid product increased with increasing reaction temperature. The germination index (GI), a measure of the percentage of germination, exceeded 90 % at 125, 150, and 175 °C in diluted samples, while it decreased to 18 % at 200 °C. Although a longer retention time contributed to an increase in TOC of liquid products, it did not increase the GI values. The liquid product should be diluted or adjusted before use as fertilizer to prevent phytotoxicity. In our analysis of ARB and ARGs, E. coli and antibiotic-resistant E. coli were completely reduced after HTT, except for the operating conditions of 125 °C and 60 min. Although both a higher reaction temperature and longer retention time tended to be better for the reduction of ARGs and intI1, it was found that the longer retention time is much more effective than the higher reaction temperature. The reduction of target ARGs and intI1 was 2.9-log under175 °C and 120 min. Comprehensively considering the fertilizer potential of liquid product and the reduction of ARB and ARGs, 175 °C of reaction temperature and 120 min of retention time of operating conditions for HTT were recommended.

Effect of composting and storage on the microbiome and resistome of cattle manure from a commercial dairy farm in Poland.

Manure from food-producing animals, rich in antibiotic-resistant bacteria and antibiotic resistance genes (ARGs), poses significant environmental and healthcare risks. Despite global efforts, most manure is not adequately processed before use on fields, escalating the spread of antimicrobial resistance. This study examined how different cattle manure treatments, including composting and storage, affect its microbiome and resistome. The changes occurring in the microbiome and resistome of the treated manure samples were compared with those of raw samples by high-throughput qPCR for ARGs tracking and sequencing of the V3-V4 variable region of the 16S rRNA gene to indicate bacterial community composition. We identified 203 ARGs and mobile genetic elements (MGEs) in raw manure. Post-treatment reduced these to 76 in composted and 51 in stored samples. Notably, beta-lactam, cross-resistance to macrolides, lincosamides and streptogramin B (MLSB), and vancomycin resistance genes decreased, while genes linked to MGEs, integrons, and sulfonamide resistance increased after composting. Overall, total resistance gene abundance significantly dropped with both treatments. During composting, the relative abundance of genes was lower midway than at the end. Moreover, higher biodiversity was observed in samples after composting than storage. Our current research shows that both composting and storage effectively reduce ARGs in cattle manure. However, it is challenging to determine which method is superior, as different groups of resistance genes react differently to each treatment, even though a notable overall reduction in ARGs is observed.

Devitalization of Bacteria in Composted Cattle Manure with Natural Additives and Risk for Environment.

Nowadays, there is an effort to improve the effectiveness of the composting process, supported by the addition of various supplements to reduce soil nutrition losses and increase soil remediation. The aim of this study was to examine the devitalization effect of natural additives like zeolite-clinoptilolite and its combination with hydrated lime in composted cattle manure on indicator and pathogen bacteria. The composting process was running in three static piles of cattle manure mixed with wheat straw (control, zeolite-lime, and zeolite) for 126 days. Composted manure substrates were determined for physicochemical (temperature, pH, nitrogen and phosphorus content, C/N, organic matter, and moisture) and microbiological analyses (Salmonella spp., indicator bacteria). The effects of additives were reflected in changes in physicochemical factors, e.g., an increase in temperature (<53 °C) or pH (<9.3). According to Pearson correlation, these changes (pH, Nt, Pt) resulted in a significant decrease (p < 0.001) of indicator bacteria (two or three orders) in zeolite pile or zeolite-lime pile. Die-off of Salmonella spp. in the zeolite-lime pile was indicated within 41 days; in other piles, this occurred on day 63. Our results can aid in further optimizing the composting of cattle manure in order to lower environmental pollution and the risk of human infection.

Composting reduces the risks of resistome in beef cattle manure at the transcriptional level.

Transcriptomic evidence is needed to determine whether composting is more effective than conventional stockpiling in mitigating the risk of resistome in livestock manure. The objective of this study is to compare composting and stockpiling for their effectiveness in reducing the risk of antibiotic resistance in beef cattle manure. Samples collected from the center and the surface of full-size manure stockpiling and composting piles were subject to metagenomic and metatranscriptomic analyses. While the distinctions in resistome between stockpiled and composted manure were not evident at the DNA level, the advantages of composting over stockpiling were evident at the transcriptomic level in terms of the abundance of antibiotic resistance genes (ARGs), the number of ARG subtypes, and the prevalence of high-risk ARGs (i.e., mobile ARGs associated with zoonotic pathogens). DNA and transcript contigs show that the pathogen hosts of high-risk ARGs included Escherichia coli O157:H7 and O25b:H4, Klebsiella pneumoniae, and Salmonella enterica. Although the average daily temperatures for the entire composting pile exceeded 55°C throughout the field study, more ARG and ARG transcripts were removed at the center of the composting pile than at the surface. This work demonstrates the advantage of composting over stockpiling in reducing ARG risk in active populations in beef cattle manure.IMPORTANCEProper treatment of manure before land application is essential to mitigate the spread of antibiotic resistance in the environment. Stockpiling and composting are two commonly used methods for manure treatment. However, the effectiveness of composting in reducing antibiotic resistance in manure has been debated. This work compared the ability of these two methods to reduce the risk of antibiotic resistance in beef cattle manure. Our results demonstrate that composting reduced more high-risk resistance genes at the transcriptomic level in cattle manure than conventional stockpiling. This finding not only underscores the effectiveness of composting in reducing antibiotic resistance in manure but also highlights the importance of employing RNA analyses alongside DNA analyses.

Effects of Antibiotics on the DAMO Process and Microbes in Cattle Manure.

Denitrifying anaerobic methane oxidation (DAMO) can mitigate methane emissions; however, this process has not been studied in cattle manure, an important source of methane emissions in animal agriculture. The objective of this study was to investigate the occurrence of DAMO microbes in cattle manure and examine the impacts of veterinary antibiotics on the DAMO process in cattle manure. Results show that DAMO archaea and bacteria consistently occur at high concentrations in beef cattle manure. During the long-term operation of a sequencing batch reactor seeded with beef cattle manure, the DAMO activities intensified, and DAMO microbial biomass increased. Exposure to chlortetracycline at initial concentrations up to 5000 µg L-1 did not inhibit DAMO activities or affect the concentrations of the 16S rRNA gene and functional genes of DAMO microbes. In contrast, exposure to tylosin at initial concentrations of 50 and 500 µg L-1 increased the activities of the DAMO microbes. An initial concentration of 5000 µg L-1 TYL almost entirely halted DAMO activities and reduced the concentrations of DAMO microbes. These results show the occurrence of DAMO microbes in cattle manure and reveal that elevated concentrations of dissolved antibiotics could inhibit the DAMO process, potentially affecting net methane emissions from cattle manure.

Study of the variation law of the airflow resistance and related physical parameters of the pile throughout compost process with cattle manure.

Composting is widely adopted in livestock waste management, and the ventilation system control is essential for composting efficiency. For ventilation system, the airflow resistance is a major factor influencing the ventilation intensity and oxygen supply capacity. This study explored the variation law of airflow resistance, bulk density, specific gravity, particle size and total pressure throughout composting with cattle manure. The airflow resistance was calculated with Ergun equation, and contribution coefficients of different components were analyzed with principal component analysis (PCA). Results showed that the viscous airflow resistance was dominant throughout cattle manure composting. The average airflow resistance was 0.146 Pa/m, and resistance of pile at lower layer was higher than that at the upper layer by 18.1 %. For contribution coefficient affecting airflow resistance, the ranks were bulk density, average particle size and specific gravity. During composting process, the average airflow resistance decreased by 40.1 % and the total pressure reduced by 3.47 %. All parameters had the greatest variation at thermophilic phase, which accounted for more than 60 % of the total variation amplitude. Meanwhile, less than 10 % of the total pressure was used to overcome the airflow resistance. Therefore, reducing bulk density of pile should be considered preferentially to decrease the airflow resistance. During cattle manure composting process, the total pressure of ventilation system ought to be adjusted with the aerobic reaction to a lower level, especially at thermophilic phase with the most rapid descent rate. This study can provide support for reducing the energy consumption required for ventilation of composting.

Soil-specific responses in the antibiotic resistome of culturable Acinetobacter spp. and other non-fermentative Gram-negative bacteria following experimental manure application.

Acinetobacter spp. and other non-fermenting Gram-negative bacteria (NFGNB) represent an important group of opportunistic pathogens due to their propensity for multiple, intrinsic, or acquired antimicrobial resistance (AMR). Antimicrobial resistant bacteria and their genes can spread to the environment through livestock manure. This study investigated the effects of fresh manure from dairy cows under antibiotic prophylaxis on the antibiotic resistome and AMR hosts in microcosms using pasture soil. We specifically focused on culturable Acinetobacter spp. and other NFGNB using CHROMagar Acinetobacter. We conducted two 28-days incubation experiments to simulate natural deposition of fresh manure on pasture soil and evaluated the effects on antibiotic resistance genes (ARGs) and bacterial hosts through shotgun metagenomics. We found that manure application altered the abundance and composition of ARGs and their bacterial hosts, and that the effects depended on the soil source. Manure enriched the antibiotic resistome of bacteria only in the soil where native bacteria had a low abundance of ARGs. Our study highlights the role of native soil bacteria in modulating the consequences of manure deposition on soil and confirms the potential of culturable Acinetobacter spp. and other NFGNB to accumulate AMR in pasture soil receiving fresh manure.

Long-term impacts of conservation pasture management in manuresheds on system-level microbiome and antibiotic resistance genes.

Animal manure improves soil fertility and organic carbon, but long-term deposition may contribute to antibiotic resistance genes (ARGs) entering the soil-water environment. Additionally, long-term impacts of applying animal manure to soil on the soil-water microbiome, a crucial factor in soil health and fertility, are not well understood. The aim of this study is to assess: (1) impacts of long-term conservation practices on the distribution of ARGs and microbial dynamics in soil, and runoff; and (2) associations between bacterial taxa, heavy metals, soil health indicators, and ARGs in manures, soils, and surface runoff in a study following 15 years of continuous management. This management strategy consists of two conventional and three conservation systems, all receiving annual poultry litter. High throughput sequencing of the 16S ribosomal RNA was carried out on samples of cattle manure, poultry litter, soil, and runoff collected from each manureshed. In addition, four representative ARGs (intl1, sul1, ermB, and blactx-m-32) were quantified from manures, soil, and runoff using quantitative PCR. Results revealed that conventional practice increased soil ARGs, and microbial diversity compared to conservation systems. Further, ARGs were strongly correlated with each other in cattle manure and soil, but not in runoff. After 15-years of conservation practices, relationships existed between heavy metals and ARGs. In the soil, Cu, Fe and Mn were positively linked to intl1, sul1, and ermB, but trends varied in runoff. These findings were further supported by network analyses that indicated complex co-occurrence patterns between bacteria taxa, ARGs, and physicochemical parameters. Overall, this study provides system-level linkages of microbial communities, ARGs, and physicochemical conditions based on long-term conservation practices at the soil-water-animal nexus.

Evaluation of ammonia emission reducing effect by adding waste cooking oil in pilot-scale composting of dairy cattle manure.

OBJECTIVE: In our previous study, we observed that the addition of waste cooking oil (WCO) reduced ammonia (NH3) emissions during laboratory-scale composting of dairy cattle manure under low-aeration condition. Therefore, this study aimed to evaluate the effect of addition of WCO on NH3 emissions reduction during pilot-scale composting of dairy cattle manure, which is close to the conditions of practical composting treatment. METHODS: Composting tests were conducted using pilot-scale composting facilities (1.8 m3 of capacity). The composting mixtures were prepared from manure, sawdust, and WCO. Two treatments were set: without WCO (Control) and with WCO added to 3 wt% of manure (WCO3). Composting was conducted under continuous aeration at 40 L/min, corresponding to 22.2 L/(min‧m3) of the mixture at the start of composting. The changes in temperatures, NH3 concentrations in the exhaust gases, and contents of the composted mixtures were analyzed. Based on these analysis results, the effect of WCO addition on NH3 emissions and nitrogen loss during composting was evaluated. RESULTS: During composting, the temperature increase of the composting mixture became higher, and the decreases of weight and water content of the mixture became larger in WCO3 than in Control. In the decrease of weight, and the residual weight and water content of the mixture, significant differences (p<0.05) were detected between the two treatments at the end of composting. The NH3 concentrations in the exhaust gases tended to be lower in WCO3 than in Control. Nitrogen loss was 21.5% lower in WCO3 than in Control. CONCLUSION: Reduction of NH3 emissions by the addition of WCO under low aeration condition was observed in pilot-scale composting, as well as in laboratory-scale composting. This result suggests that this method is effective in reducing NH3 emissions in practicalscale composting.

Modelling the deactivation of Escherichia coli in Nigerian soils amended with differently treated manures.

AIM: This study aimed to simulate deactivation of Escherichia coli in soils amended with cattle manure after burning, anaerobic digestion, composting, or without treatment. METHOD AND RESULTS: The Weibull survival function was used to describe deactivation of E. coli. Parameters for each treatment were determined using E. coli measurements from manure-amended soils and evaluated against measurements at different application rates. A statistically significant correlation and high coincidence between the simulated and measured values were obtained. The simulations revealed that although anaerobic digestion or burning of cattle manure effectively reduced the E. coli loads to background levels, burning retained very little nitrogen, so the ash residue was ineffective as an organic fertilizer. Anaerobic digestion was most effective at reducing E. coli levels while retaining a high proportion of N in the bioslurry residue, but the persistence of E. coli was higher than in compost. CONCLUSION: The results from this study suggest that the safest method for production of organic fertilizer would involve anaerobic digestion to reduce E. coli, followed by composting to reduce its persistence.

Differential impacts of salinity on antibiotic resistance genes during cattle manure stockpiling are linked to mobility potentials revealed by metagenomic sequencing.

Livestock manure is an important source of antibiotic resistance genes (ARGs), and its salinity level can change during stockpiling. To understand how the salinity changes affect the fate of ARGs, cattle manure was adjusted of salinity and stockpiled in laboratory microcosms at low (0.3% salt), moderate (3.0%) and high salinity levels (10.0%) for 44 days. Amongst the five ARGs (tetO, blaTEM, sul1, tetM, and ermB) and the first-class integrase (intI1) monitored by qPCR, the relative abundance of tetO and blaTEM exhibited no clear trend in response to salinity levels, while that of sul1, tetM, ermB and intI1 showed clear downward trends over time at the lower salinity levels (0.3% and 3%) but not at the high salinity level (10%). Metagenomic contig construction of cattle manure samples revealed that sul1, tetM and ermB genes were more likely to associate with mobile genetic elements (MGEs) than tetO and blaTEM, suggesting that their slower decay at higher salinity levels was either caused by horizontal gene transfer or co-selection of ARGs and osmotic stress resistant determinants. Further analysis of metagenomic contigs showed that osmotic stress resistance can also be located on MGEs or in conjunction with ARGs.

Graphene nano zinc oxide reduces the expression and release of antibiotic resistance-related genes and virulence factors in animal manure.

Animal manure contains many antibiotic resistance genes (ARGs) and virulence factors (VFs), posing significant health threats to humans. However, the effects of graphene nano zinc oxide (GZnONP), a zinc bioaugmentation substitute, on bacterial chemotaxis, ARGs, and VFs in animal manure remain scanty. Herein, the effect of GZnONP on the in vivo anaerobic expression of ARGs and VFs in cattle manure was assessed using high-throughput sequencing. Results showed that GZnONP inhibited bacterial chemotaxis by reducing the zinc pressure under anaerobic fermentation, altering the microbial community structure. The expression of ARGs was significantly lower in GZnONP than in zinc oxide and nano zinc oxide (ZnONP) groups. The expression of VFs was lower in the GZnONP than in the zinc oxide and ZnONP groups by 9.85 % and 13.46 %, respectively. Co-occurrence network analysis revealed that ARGs and VFs were expressed by the Spirochaetes phylum, Paraprevotella genus, and Treponema genus et al. The ARGs-VFs coexistence was related to the expression/abundance of ARGs and VFs genes. GZnONP reduces the abundance of certain bacterial species by disrupting chemotaxis, minimizing the transfer of ARGs and VFs. These findings suggest that GZnONP, a bacterial chemotaxis suppressor, effectively reduces the expression and release of ARGs and VFs in animal manure.

Effects of hydrothermal pretreatment on methane potential of anaerobic digestion sludge cake of cattle manure containing sawdust as bedding materials.

OBJECTIVE: The purpose of this study was to analyze the effect of the hydrothermal pretreat-ment of anaerobic digestion sludge cake (ADSC) of cattle manure on the solubilization of organic matter and the methane yield to improve the anaerobic digestion efficiency of cattle manure collected from the sawdust pens of cattle. METHODS: Anaerobic digestion sludge cake of cattle manure was thermally pretreated at 160°C, 180°C, 200°C, and 220°C by a hydrothermal pressure reactor, and the biochemical methane potential of ADSC hydrolysate was analyzed. Methane yield recovered by the hydrothermal pretreatment of ADCS was estimated based on mass balance. RESULTS: The chemical oxygen demand solubilization degree (CODs) of the hydrothermal hydrolysate increased to 63.56%, 67.13%, 70.07%, and 66.14% at the hydrothermal reaction temperatures of 160°C, 180°C, 200°C, and 220°C, respectively. Considering the volatile solids content obtained after the hydrothermal pretreatment, the methane of 10.2 Nm3/ton-ADSC was recovered from ADSC of 1.0 ton, and methane yields of ADSC hydrolysate increased to 15.6, 18.0, 17.4, and 17.2 Nm3/ton-ADSC. CONCLUSION: Therefore, the optimal hydrothermal reaction temperature that yielded the maximum methane yield was 180°C based on mass balance, and the methane yield from cattle manure containing sawdust was improved by the hydrothermal pretreatment of ADSC.

Digestate from Agricultural Biogas Plants as a Reservoir of Antimicrobials and Antibiotic Resistance Genes-Implications for the Environment.

Antimicrobials and antibiotic resistance genes (ARGs) in substrates processed during anaerobic digestion in agricultural biogas plants (BPs) can reach the digestate (D), which is used as fertilizer. Antimicrobials and ARGs can be transferred to agricultural land, which increases their concentrations in the environment. The concentrations of 13 antibiotics in digestate samples from biogas plants (BPs) were investigated in this study. The abundance of ARGs encoding resistance to beta-lactams, tetracyclines, sulfonamides, fluoroquinolones, macrolide-lincosamide-streptogramin antibiotics, and the integrase genes were determined in the analyzed samples. The presence of cadmium, lead, nickel, chromium, zinc, and mercury was also examined. Antimicrobials were not eliminated during anaerobic digestion. Their concentrations differed in digestates obtained from different substrates and in liquid and solid fractions (ranging from 62.8 ng/g clarithromycin in the solid fraction of sewage sludge digestate to 1555.9 ng/L doxycycline in the liquid fraction of cattle manure digestate). Digestates obtained from plant-based substrates were characterized by high concentrations of ARGs (ranging from 5.73 × 102 copies/gDcfxA to 2.98 × 109 copies/gDsul1). The samples also contained mercury (0.5 mg/kg dry mass (dm)) and zinc (830 mg/kg dm). The results confirmed that digestate is a reservoir of ARGs (5.73 × 102 to 8.89 × 1010 copies/gD) and heavy metals (HMs). In addition, high concentrations of integrase genes (105 to 107 copies/gD) in the samples indicate that mobile genetic elements may be involved in the spread of antibiotic resistance. The study suggested that the risk of soil contamination with antibiotics, HMs, and ARGs is high in farms where digestate is used as fertilizer.

Potential of vermicomposting with mixtures of animal manure and vegetable leaves in the development of Eisenia foetida, microbial biomass, and enzymatic activity under semi-arid conditions.

Vermicomposting is the bio-oxidation and stabilization of organic matter involving relationships between the action of earthworms and microorganisms and the activation and dynamics of several enzyme activities. Semi-arid farmers to make (extra) money and organic production, produce their vermicompost using plant residues and animal manure, but there is no information about the final product generated. Thus, this study aimed to analyze the potential of vermicomposting with mixtures of animal manure and vegetable leaves in the development of Eisenia foetida, microbial biomass, and enzymatic activity in the semi-arid region, Brazil. The experimental design applied was randomized block in a 6 × 4 factorial scheme with four replicates, with six treatments (mixtures of cattle manure, goat manure, cashew leaves, and catanduva leaves) and evaluated at four-time intervals (30, 60, 90, and 120 days of vermicomposting). The treatments were placed in polyethylene pots in the same site, environmental conditions, and residues proportions as used by farmers. The characteristics analyzed were the number of earthworms (NE), total earthworm biomass (TEB) and earthworm multiplication index (MI), microbial biomass carbon (MBC), and activities of enzymes ß-glucosidase, dehydrogenase, alkaline and acid phosphatases. The cattle manure vermicomposted shows the highest average values observed for NE, MI, TEB, MBC, and enzymatic activity, regardless of the plant leaves mix. In general, the enzymes activities were found in the descending order of ß-glucosidase > alkaline phosphatase > dehydrogenase > acid phosphatase. The maturation dynamics of vermicompost were characterized by a decline in the microbial population and number and biomass of earthworms in the substrate and consequently a decrease in new enzyme synthesis and degradation of the remaining enzyme pool. Microbial biomass and enzymatic activity were indicators for changes in the quality of vermicompost.