Community coalescence and plant host filtering determine the spread of tetracycline resistance genes from pig manure into the microbiome continuum of the soil-plant system.

The spread of livestock manure-borne antibiotic resistance genes (ARGs) into agroecosystems through manure application poses a potential threat to human health. However, there is still a knowledge gap concerning ARG dissemination in coalescing manure, soil and plant microbiomes. Here, we examined the fate of tetracycline resistance genes (TRGs) originating from pig manure microbiomes and spread in the soil-A thaliana system and explored the effects of microbial functions on TRGs spread at different interfaces. Our results indicate that the TRGs abundances in all microbiome continuum of the soil-A. thaliana system were significantly increased with the application of a living manure microbiome, although the addition of manure with both an active and inactive microbiome caused a shift in the microbial community composition. This was attributed to the increasing relative abundances of tetA, tetL, tetM, tetO, tetW and tolC in the system. The application of living manure with DOX residues resulted in the highest relative abundance of total TRGs (3.30×10-3 copies/16S rRNA gene copies) in the rhizosphere soil samples. Community coalescence of the manure and soil microbiomes increased the abundance of Firmicutes in the soil and root microbiome, which directly explains the increase in TRG abundance observed in these interfaces. In contrast, the leaf microbiome differed markedly from that of the remaining samples, indicating strong plant host filtering effects on Firmicutes and TRGs from pig manure. The random forest machine learning model revealed microbial functions and their significant positive correlation with TRG abundance in the microbiome continuum of the system. Our findings revealed that community coalescence is the main driver of TRG spread from manure to the soil and root microbiomes. Plant host filtering effects play a crucial role in allowing certain microbial groups to occupy ecological niches in the leaves, thereby limiting the establishment of manure-borne TRGs in aboveground plant tissues.

Development of an effective one-step double-antigen sandwich ELISA based on p72 to detect antibodies against African swine fever virus.

African swine fever (ASF), caused by ASF virus (ASFV), is a highly contagious and lethal disease of domestic pigs leading to tremendous economic losses. As there are no vaccines and drugs available. An effective diagnosis to eliminate ASFV-infected pigs is a crucial strategy to prevent and control ASF. To this end, ASFV capsid protein p72 was expressed using Chinese hamster ovary (CHO) cells and subsequently conjugated with horseradish peroxidase (HRP) to develop a one-step double-antigen sandwich enzyme-linked immunosorbent assay (one-step DAgS-ELISA). The performance of this ELISA for detecting ASFV antibodies was evaluated. Overall, a diagnostic sensitivity of 97.96% and specificity of 98.96% was achieved when the cutoff value was set to 0.25. No cross-reaction with healthy pig serum and other swine viruses was observed. The coefficients of variation of the intra-assay and inter-assay were both <10%. Importantly, this ELISA could detect antibodies in standard serum with 12,800-fold dilution, and seroconversion started from the 7th day post-inoculation (dpi), showing excellent analytical sensitivity and great utility. Furthermore, compared to the commercial kit, this ELISA had a good agreement and significantly shorter operation time. Collectively, a novel one-step DAgS-ELISA for detecting antibodies against ASFV is developed, which will be reliable and convenient to monitor ASFV infection.

Screening of linear B-cell epitopes and its proinflammatory activities of Haemophilus parasuis outer membrane protein P2.

Haemophilus parasuis is a commensal organism of the upper respiratory tract of pigs, but virulent strains can cause Glässer's disease, resulting in significant economic losses to the swine industry. OmpP2 is an outer membrane protein of this organism that shows considerable heterogeneity between virulent and non-virulent strains, with classification into genotypes I and II. It also acts as a dominant antigen and is involved in the inflammatory response. In this study, 32 monoclonal antibodies (mAbs) against recombinant OmpP2 (rOmpP2) of different genotypes were tested for reactivity to a panel of OmpP2 peptides. Nine linear B cell epitopes were screened, including five common genotype epitopes (Pt1a, Pt7/Pt7a, Pt9a, Pt17, and Pt19/Pt19a) and two groups of genotype-specific epitopes (Pt5 and Pt5-II, Pt11/Pt11a, and Pt11a-II). In addition, we used positive sera from mice and pigs to screen for five linear B-cell epitopes (Pt4, Pt14, Pt15, Pt21, and Pt22). After porcine alveolar macrophages (PAMs) were stimulated with overlapping OmpP2 peptides, we found that the epitope peptides Pt1 and Pt9, and the loop peptide Pt20 which was adjacent epitopes could all significantly upregulated the mRNA expression levels of IL-1α, IL-1ß, IL-6, IL-8, and TNF-α. Additionally, we identified epitope peptides Pt7, Pt11/Pt11a, Pt17, Pt19, and Pt21 and loop peptides Pt13 and Pt18 which adjacent epitopes could also upregulate the mRNA expression levels of most proinflammatory cytokines. This suggested that these peptides may be the virulence-related sites of the OmpP2 protein, with proinflammatory activity. Further study revealed differences in the mRNA expression levels of proinflammatory cytokines, including IL-1ß and IL-6, between genotype-specific epitopes, which may be responsible for pathogenic differences between different genotype strains. Here, we profiled a linear B-cell epitope map of the OmpP2 protein and preliminarily analyzed the proinflammatory activities and effects of these epitopes on bacterial virulence, providing a reliable theoretical basis for establishing a method to distinguish strain pathogenicity and to screen candidate peptides for subunit vaccines.

Prevalence and genetic diversity of porcine circovirus type 2 in northern Guangdong Province during 2016-2021.

The emergence and widespread of porcine circovirus-associated diseases (PCVADs), mainly caused by porcine circovirus type 2 (PCV2), threatens the Chinese swine industry. In this study, to investigate the recent prevalence of PCV2 in northern Guangdong Province of China, 573 tissue samples from 132 pig farms were collected during 2016-2021 and analyzed via PCR. Overall, 51.38% (297/573, 95%CI 47.74-55.92) samples were tested PCV2 positive. The detection rate of PCV2 was significantly lower in samples collected before 2016-2018 than after the outbreak of African Swine Fever (2019-2021), being 59.85% (158/264, 95%CI 53.94-65.76) and 41.47% (141/340, 95%CI 36.43-46.71), respectively. On the other end, the genetic characteristics of 26 PCV2 strains were further analyzed. These PCV2 strains belonged to three genotypes, including PCV2a, PCV2b, and PCV2d. Specifically, the predominant genotype prevalent during two periods (2016-2018 and 2019-2021) wasPCV2b (81.82%, 9/11) and PCV2d (80.0%, 12/15), respectively. The results above illustrated the high prevalence and the genetic evolution feature of PCV2 in Guangdong Province in recent years.

The construction of a single-crystalline SbSI nanorod array-WO3 heterostructure photoanode for high PEC performance.

A novel kind of highly efficient photoanode was constructed with a SbSI/WO3 heterostructurefabricated through two hydrothermal reactions followed by an iodination reaction (WO3 → Sb2S3/WO3 → SbSI/WO3). After optimizing the solvent [carbon disulfide (CS2)] for SbI3, the SbSI(CS2)/WO3 photoanode shows high-density single-crystalline SbSI nanorods growing along the polar [001] direction on WO3 nanoplates, resulting in excellent photocurrent performance (∼2.1 mA cm-2@1.23 V vs. RHE) and an improved photostability. It is evidenced that the higher crystallinity of SbSI has a positive effect on the photostability of the constructed SbSI/WO3 photoanodes.

A novel approach to realize SANI process in freshwater sewage treatment--Use of wet flue gas desulfurization waste streams as sulfur source.

SANI (Sulfate reduction, Autotrophic denitrification and Nitrification Integrated) process has been approved to be a sludge-minimized sewage treatment process in warm and coastal cities with seawater supply. In order to apply this sulfur-based process in inland cold areas, wet flue gas desulfurization (FGD) can be simplified and integrated with SANI process, to provide sulfite as electron carrier for sulfur cycle in sewage treatment. In this study, a lab-scale system of the proposed novel process was developed and run for over 200 days while temperature varied between 30 and 5 °C, fed with synthetic FGD wastewaters and sewage. The sulfite-reducing upflow anaerobic sludge bed (SrUASB) reactor, as the major bioreactor of the system, removed 86.9% of organics while the whole system removed 94% of organics even when water temperature decreased to around 10 °C. The bactericidal effect of sulfite was not observed in the SrUASB reactor, while thiosulfate was found accumulated under psychrophilic conditions. The sludge yield of the SrUASB reactor was determined to be 0.095 kg VSS/kg COD, higher than of sulfate reduction process but still much lower than of conventional activated sludge processes. The dominant microbes in the SrUASB reactor were determined as Lactococcus spp. rather than sulfate-reducing bacteria, but sulfite reduction still contributed 85.5% to the organic carbon mineralization in this reactor. Ammonia and nitrate were effectively removed in the aerobic and anoxic filters, respectively. This study confirms the proposed process was promising to achieve sludge-minimized sewage treatment integrating with flue gas desulfurization in inland and cold areas.

Effect of volatile emissions, especially alpha-pinene, from persimmon trees infested by Japanese wax scales or treated with methyl jasmonate on recruitment of ladybeetle predators.

Persimmon, Diospyros kaki L., is an important fruit tree in northern China. Japanese wax scale, Ceroplastes japonicus Green (Hemiptera: Coccoidea: Coccidae), is the most destructive pest in persimmon orchards and is difficult to control using chemical pesticides. This paper studied the variety of volatile emissions from persimmon trees attacked by wax scale or induced by applications of methyl jasmonate (MeJA), an exogenous signaling material. We also studied the role of volatiles in recruiting the ladybeetle, Chilocorus kuwanae Silvestri (Coleoptera: Coccinellidae), a primary predator of wax scale. The results showed that the ladybeetles displayed strong taxis responses to the odor sources from trees treated with MeJA and wax scale-damaged trees. Within 24 h after the trees were treated with MeJA, the number of ladybeetles responding to the treated trees fluctuated based on the amount of volatiles released over a diel period. Chemical analysis of the volatiles showed that terpenoid compounds, especially alpha-pinene, increased from both the wax scale-damaged and MeJA-treated trees and that this was the reason for the ladybeetle attraction. Therefore, alpha-pinene was used as a single signal source to examine the taxis response of C. kuwanae. The results suggest that alpha-pinene plays a significant role in attracting the ladybeetles. It is therefore concluded that MeJA application might be used to stimulate the defense function of persimmon trees by inducing the release of terpenoids, especially alpha-pinene, and thereby recruiting more ladybeetles to control the scale insects.