A global phosphoproteomics analysis of adult Fasciola gigantica by LC-MS/MS.

Protein phosphorylation plays key roles in a variety of essential cellular processes. Fasciola gigantica is a tropical liver fluke causing hepatobiliary disease fascioliasis, leading to human health threats and heavy economic losses. Although the genome and protein kinases of F. gigantica provided new insights to understand the molecular biology and etiology of this parasite, there is scant knowledge of protein phosphorylation events in F. gigantica. In this study, we characterized the global phosphoproteomics of adult F. gigantica by phosphopeptide enrichment-based LC-MS/MS, a high-throughput analysis to maximize the detection of a large repertoire of phosphoproteins and phosphosites. A total of 1030 phosphopeptides with 1244 phosphosites representing 635 F. gigantica phosphoproteins were identified. The phosphoproteins were involved in a wide variety of biological processes including cellular, metabolic, and single-organism processes. Meanwhile, these proteins were found predominantly in cellular components like membranes and organelles with molecular functions of binding (51.3%) and catalytic activity (40.6%). The KEGG annotation inferred that the most enriched pathways of the phosphoproteins included tight junction, spliceosome, and RNA transport (each one contains 15 identified proteins). Combining the reports in other protozoa and helminths, the phosphoproteins identified in this work play roles in metabolic regulation and signal transduction. To our knowledge, this work performed the first global phosphoproteomics analysis of adult F. gigantica, which provides valuable information for development of intervention strategies for fascioliasis.

Mineralization Reduces the Toxicity and Improves Stability and Protective Immune Response Induced by Toxoplasma gondii.

Vaccination is an ideal strategy for the control and prevention of toxoplasmosis. However, the thermostability and effectiveness of vaccines limit their application. Here, calcium mineralization was used to fabricate Toxoplasma gondii tachyzoites as immunogenic core-shell particles with improved immune response and thermostability. In the current study, T. gondii RH particles coated with mineralized shells were fabricated by calcium mineralization. The mineralized shells could maintain the T. gondii tachyzoites structural integrity for at least 12 months and weaken the virulence. Immunization of mice with mineralized tachyzoites induced high levels of T. gondii-specific antibodies and cytokines. The immunized mice were protected with a 100% survival rate in acute and chronic infection, and brain cyst burdens were significantly reduced. This study reported for the first time the strategy of calcium mineralization on T. gondii and proved that mineralized tachyzoites could play an immune protective role, thus expanding the application of biomineralization in T. gondii vaccine delivery.

Epidemiology of Fasciola spp. in the intermediate host in China: A potential risk for fasciolosis transmission.

Fasciolosis is a zoonotic disease as 600 million animals have been infected, and 180 million people are at risk of the infection in the world. Snail as the intermediate host of Fasciola is an essential and important factor in the transmission of fasciolosis, while its potential risk for transmission has not been studied. In this study, 3561 snails collected from large-scale regions of China were examined by nest-PCR method. A total of 345 snails were positive for Fasciola spp., with an overall prevalence of 9.7%. Prevalence in central and southern China, characterized by a subtropical monsoon climate, was relatively low (8.0%), while a high infection rate (36.9%) was found on the plateau area (altitude > 500 m). In combination with previous findings, the study showed a highly positive correlation between snails and animal infection in central and eastern China, thus indicating that infected snails could be an indispensable risk factor for fasciolosis transmission. Epidemiological surveillance of snails will help assess the risk of fasciolosis in humans or ruminants, which promotes future prevention of this zoonotic disease.

Removal of abamectin and conventional pollutants in vertical flow constructed wetlands with Fe-modified biochar.

To improve the ability of constructed wetlands to remove abamectin (ABM) and nutrients, the influence of four different substrates on constructed wetlands was studied. Four vertical up-flow constructed wetlands (UVCWs) were established to treat simulated agricultural wastewater: CW1 (quartz sand + pebbles), CW2 (pebbles + coke), CW3 (Fe-modified biochar + pebbles + coke), and CW4 (unmodified biochar + pebbles + coke). Under different combinations of hydraulic loading and organic loading, CW3 was extremely effective at removing nitrogen compared with CW1, CW2 and CW4. We found that CW3 was the most effective at treating ABM and conventional pollutants. The highest efficiency of removal of abamectin (99%), COD (98%), NH4 +-N (65%), and TP (80%) was obtained in CW3. These results were directly verified by microbiological tests and microbial community analysis. The microbial diversity of CW3 and CW4 was significantly higher than those of CW1 and CW2. Fe-modified biochar provides a feasible and effective amendment for constructed wetlands to improve the nitrogen removal for C/N (2.5 : 1-5 : 1) wastewater by the ability of microbes to remove nitrogen. Fe-modified bamboo charcoal can be used in engineering as a new type of green environmental protection constructed wetland filler in the future.

[Correlations Between Substrate Structure and Microbial Community in Subsurface Flow Constructed Wetlands].

To identify the microbial factors that cause the differences in the purification performance of constructed wetlands with different substrate structures, the relationship between the substrate structure and the microbial community composition in horizontal subsurface flow constructed wetlands (HSSFCWs) was studied by high throughput sequencing. The results revealed that the purification performance of a six-layer constructed wetland (CW6), of which the permeability coefficient gradually increased from the surface layer to the bottom layer, was the highest among the three constructed wetland systems. The average concentrations of COD, TN, NO3--N, and NH4+-N in the effluent were 39, 11, 0.35, and 4 mg·L-1, respectively. The monolayer structure constructed wetland (CW1) had the worst purifying efficiency, with average effluent concentrations of 95, 21, 0.60 and 12 mg·L-1 for COD, TN, NO3--N, and NH4+-N, respectively. The results of the high-throughput sequencing showed that the number of microbial OTUs in multilayer structure wetlands was slightly lower than that in the monolayer structure wetland, but the relative abundance of the dominant phylum Proteobacteria and the nitrifying and denitrifying bacteria in the genus was significantly higher than the monolayer structure wetland. The results of PCA and heatmap indicated that there were significant differences in the spatial distribution of microbes in the genus of Proteobacteria in CW3 and CW6, which facilitated the degradation of pollutants. No significant differences were found in the community structure of CW1.

[Correlation of substrate structure and hydraulic characteristics in subsurface flow constructed wetlands].

The correlation of substrate structure and hydraulic characteristics was studied by numerical simulation combined with experimental method. The numerical simulation results showed that the permeability coefficient of matrix had a great influence on hydraulic efficiency in subsurface flow constructed wetlands. The filler with a high permeability coefficient had a worse flow field distribution in the constructed wetland with single layer structure. The layered substrate structure with the filler permeability coefficient increased from surface to bottom could avoid the short-circuited flow and dead-zones, and thus, increased the hydraulic efficiency. Two parallel pilot-scale constructed wetlands were built according to the numerical simulation results, and tracer experiments were conducted to validate the simulation results. The tracer experiment result showed that hydraulic characteristics in the layered constructed wetland were obviously better than that in the single layer system, and the substrate effective utilization rates were 0.87 and 0.49, respectively. It was appeared that numerical simulation would be favorable for substrate structure optimization in subsurface flow constructed wetlands.