Deep eutectic solvents pretreatment enhances methane production from anaerobic digestion of waste activated sludge: Effectiveness evaluation and mechanism elucidation.

Anaerobic digestion (AD) is a prevalent waste activated sludge (WAS) treatment, and optimizing methane production is a core focus of AD. Two DESs were developed in this study and significantly increased methane production, including choline chloride-urea (ChCl-Urea) 390% and chloride-ethylene glycol (ChCl-EG) 540%. Results showed that ChCl-Urea mainly disrupted extracellular polymeric substances (EPS) structures, aiding in initial sludge solubilization during pretreatment. ChCl-EG, instead, induced sludge self-driven organic solubilization and enhanced hydrolysis and acidification processes during AD process. Based on the extent to which the two DESs promoted AD for methane production, the AD process can be divided into stage Ⅰ and stage Ⅱ. In stage Ⅰ, ChCl-EG promoted methanogenesis more significantly, microbiological analysis showed both DESs enriched aceticlastic methanogens-Methanosarcina. Notably, ChCl-Urea particularly influenced polysaccharide-related metabolism, whereas ChCl-EG targeted protein-related metabolism. In stage Ⅱ, ChCl-Urea was more dominant than ChCl-EG, ChCl-Urea bolstered metabolism and ChCl-EG promoted genetic information processing in this stage. In essence, this study investigated the microbial mechanism of DES-enhanced sludge methanogenesis and provided a reference for future research.

Efficient phosphorus recovery from waste activated sludge: Pretreatment with natural deep eutectic solvent and recovery as vivianite.

Recycling phosphorus from waste activated sludge (WAS) is an effective method to address the nonrenewable nature of phosphorus and mitigate environmental pollution. To overcome the challenge of low phosphorus recovery from WAS due to insufficient disintegration, a method using a citric acid-based natural deep eutectic solvent (CA-NADES) assisted with low-temperature pretreatment was proposed to efficiently release and recover phosphorus. The results of 31P nuclear magnetic resonance (NMR) confirmed that low-temperature pretreatment promoted the conversion of organic phosphorus (OP) to inorganic phosphorus (IP) and enhanced the effect of CA-NADES. Changes in the three-dimensional excitation-emission matrix (3D-EEM) and flow cytometry (FCM) indicated that the method of CA-NADES with low-temperature thermal simultaneously release IP and OP by disintegrating sludge flocs, dissolving extracellular polymeric substances (EPS) structure, and cracking cells. When 5 % (v/v) of CA-NADES was added and thermally treated at 60 °C for 30 min, 43 % of total phosphorus (TP) was released from the sludge. The concentrations of proteins and polysaccharides reached 826 and 331 mg/L, respectively, which were 6.30 and 14.43 times higher than those of raw sludge. The dewatering and settling of the sludge were also improved. Metals were either enriched in the solid phase or released into the liquid phase in small quantities (most efficiencies of less than 10 %) for subsequent clean recovery. The released phosphorus was successfully recovered as vivianite with a rate of 90 %. This study develops an efficient, green, and sustainable method for phosphorus recovery from sludge using NADES and provides new insights into the high-value conversion of sludge.

Enhancing bioavailable carbon sources and minimizing ammonia emissions in distillery sludge and distiller's grains waste co-composting through deep eutectic solvent addition.

This study introduced two deep eutectic solvents, ChCl/oxalic acid (CO) and ChCl/ethylene glycol (CE), into a 34-day co-composting process of distillery sludge and distiller's grains waste to address challenges related to NH3 emissions. The addition of DES increased dissolved organic carbon by 68% to 92%, offering more utilizable carbon for microorganisms. SYTO9/PI staining and enzyme activity tests showed the CE group had higher bacterial activity and metabolic levels during the thermophilic phase than the control. Bacterial community analysis revealed that early dominance of Lactobacillus and Lysinibacillus in CE accelerated the onset of the thermophilic phase, reduced pile pH, and significantly decreased urease production by reducing Ureibacillus. Consequently, CE treatment substantially dropped NH3 emissions by 73% and nitrogen loss by 54%. Besides, CE fostered a more abundant functional microbial community during the cooling and maturation phases, enhancing deep degradation and humification of organic matter.

Untargeted Headspace-Gas Chromatography-Ion Mobility Spectrometry in Combination with Chemometrics for Detecting the Age of Chinese Liquor (Baijiu).

This paper proposes the combination of headspace-gas chromatography-ion mobility spectrometry (HS-GC-IMS) and chemometrics as a method to detect the age of Chinese liquor (Baijiu). Headspace conditions were optimized through single-factor optimization experiments. The optimal sample preparation involved diluting Baijiu with saturated brine to 15% alcohol by volume. The sample was equilibrated at 70 °C for 30 min, and then analyzed with 200 µL of headspace gas. A total of 39 Baijiu samples from different vintages (1998-2019) were collected directly from pottery jars and analyzed using HS-GC-IMS. Partial least squares regression (PLSR) analysis was used to establish two discriminant models based on the 212 signal peaks and the 93 identified compounds. Although both models were valid, the model based on the 93 identified compounds discriminated the ages of the samples more accurately according to the goodness of fit value (R2) and the root mean square error of prediction (RMSEP), which were 0.9986 and 0.244, respectively. Nineteen compounds with variable importance for prediction (VIP) scores > 1, including 11 esters, 4 alcohols, and 4 aldehydes, played vital roles in the model established by the 93 identified compounds. Overall, we determined that HS-GC-IMS combined with PLSR could serve as a rapid and accurate method for detecting the age of Baijiu.

Characterization of Streptococcus suis isolates from the diseased pigs in China between 2003 and 2007.

The aim of this study was to illustrate the characteristics of 407 strains of Streptococcus suis (S. suis) isolated from diseased pigs in China. The results revealed that S. suis, with 56.6% of the Streptococci isolates, had replaced Streptococcus equi subsp. zooepidemicus as the predominant agent. Among the strains investigated, serotype 2 (43.2%) was most prevalent, followed by serotypes 3 (14.7%) and 4, 8, 5, 7, 1/2 (3.2-6.4%). Serotype 2 was more frequently isolated from swine with systemic infection, while serotype 3 was significantly associated with pneumonia. A high percentage of S. suis serotype 2 strains (54%) belonged to the genotype sly+ mrp+ epf+, which is a highly virulent strain, as confirmed by mice infection test. Biofilm producers only had lowly virulence compared to strains with the same genotype, indicating that biofilm could be associated with virulence but be not the character of virulent strains. The present study contributes to understanding the characteristics of Streptococcus suis and controlling Streptococcal disease in China.

Employment of Near Full-Length Ribosome Gene TA-Cloning and Primer-Blast to Detect Multiple Species in a Natural Complex Microbial Community Using Species-Specific Primers Designed with Their Genome Sequences.

It remains an unsolved problem to quantify a natural microbial community by rapidly and conveniently measuring multiple species with functional significance. Most widely used high throughput next-generation sequencing methods can only generate information mainly for genus-level taxonomic identification and quantification, and detection of multiple species in a complex microbial community is still heavily dependent on approaches based on near full-length ribosome RNA gene or genome sequence information. In this study, we used near full-length rRNA gene library sequencing plus Primer-Blast to design species-specific primers based on whole microbial genome sequences. The primers were intended to be specific at the species level within relevant microbial communities, i.e., a defined genomics background. The primers were tested with samples collected from the Daqu (also called fermentation starters) and pit mud of a traditional Chinese liquor production plant. Sixteen pairs of primers were found to be suitable for identification of individual species. Among them, seven pairs were chosen to measure the abundance of microbial species through quantitative PCR. The combination of near full-length ribosome RNA gene library sequencing and Primer-Blast may represent a broadly useful protocol to quantify multiple species in complex microbial population samples with species-specific primers.

Antimicrobial susceptibility, tetracycline and erythromycin resistance genes, and multilocus sequence typing of Streptococcus suis isolates from diseased pigs in China.

Streptococcus suis (S. suis) is an emerging zoonotic pathogen causing significant economic losses in the swine industry. Here, we investigated the antimicrobial susceptibility, associated antibiotic-resistant determinants and sequence type (ST) of S. suis isolates from diseased pigs in China from 2008 to 2010. Serotype 2 was the most frequently observed strain (n=95) among the 106 S. suis strains collected, followed by serotypes 3 (n=3), 5 (n=3), 4 (n=2), 7 (n=1), 11 (n=1) and 28 (n=1). Multilocus sequence typing analysis revealed that ST1 (n=21) and ST7 (n=74) were the predominant STs, and serotype 2 was found to be significantly correlated with ST7 (P=0.017, Fisher's exact test) and CC1 (P=0.024, Fisher's exact test). The antimicrobial susceptibility results indicated that the antibiotic resistance rate was highest for tetracycline (99.1%), followed by azithromycin (68.9%), erythromycin (67.9%), clindamycin (67.9%), trimethoprim/sulfamethoxazole (16%), levofloxacin (2.8%), chloramphenicol (1.9%), cefaclor (0.9%) and ceftriaxone (0.9%). Antibiotic-resistant genes tet(M), tet(O), tet(O/W/32/O), tet(O/32/O), tet(S), tet(W), tet(L), tet(40), erm(B), mef(A/E) and msr(D) could be detected, and several tandem organizations of antibiotic resistance genes were also found in this study. In conclusion, S. suis strains isolated from diseased pigs in China were less diverse and multi-drug resistant.

Evaluation of the protective efficacy of a newly identified immunogenic protein, HP0272, of Streptococcus suis.

Streptococcus suis serotype 2 (SS2) infection is a major cause of sudden death in pigs and is of concern for humans as it has strong zoonotic capabilities. Developing novel effective vaccines would be beneficial to control SS2 infection. HP0272 is a novel immunogenic surface protein; its protective efficacy remains to be evaluated. The present mouse model found that the purified recombinant HP0272 could elicit a significant humoral antibody response, and to confer complete protection against a lethal dose of SS2 infection. In addition, real-time PCR confirmed that in vivo-induced antigen existed in most SS2 field pathogenic strains, and in half of all reference strains of different serotypes of S. suis. The results indicate that HP0272 has the potential as an effective component of a new vaccine.