Greenhouse gases emissions from aquaculture ponds: Different emission patterns and key microbial processes affected by increased nitrogen loading.

Global aquaculture production is expected to rise to meet the growing demand for food worldwide, potentially leading to increased anthropogenic greenhouse gases (GHG) emissions. As the demand for fish protein increases, so will stocking density, feeding amounts, and nitrogen loading in aquaculture ponds. However, the impact of GHG emissions and the underlying microbial processes remain poorly understood. This study investigated the GHG emission characteristics, key microbial processes, and environmental drivers underlying GHG emissions in low and high nitrogen loading aquaculture ponds (LNP and HNP). The N2O flux in HNP (43.1 ± 11.3 µmol m-2 d-1) was significantly higher than in LNP (-11.3 ± 25.1 µmol m-2 d-1), while the dissolved N2O concentration in HNP (52.8 ± 7.1 nmol L-1) was 150 % higher than in LNP (p < 0.01). However, the methane (CH4) and carbon dioxide (CO2) fluxes and concentrations showed no significant differences (p > 0.05). N2O replaced CH4 as the main source of Global Warming Potential in HNP. Pond sediments acted as a sink for N2O but a source for CH4 and CO2. The △N2O/(△N2O + â–³N2) in HNP (0.015 ± 0.007 %) was 7.7-fold higher than in LNP (0.002 ± 0.001 %) (p < 0.05). The chemical oxygen demand to NO2-N ratio was the most important environmental factor explaining the variability of N2O fluxes. Ammonia-oxidizing bacteria driven nitrification in water was the predominant N2O source, while comammox-driven nitrification and nosZII-driven N2O reduction in water were key processes for reducing N2O emission in LNP but decreased in HNP. The strong CH4 oxidization by Methylocystis and CO2 assimilation by algae resulted in low CH4 emissions and CO2 sink in the aquaculture pond. The Mantel test indicated that HNP increased N2O fluxes mainly through altering functional genes composition in water and sediment. Our findings suggest that there is a significant underestimation of N2O emissions without considering the significantly increased △N2O/(△N2O + â–³N2) caused by increased nitrogen loading.

Impact and microbial mechanism of continuous nanoplastics exposure on the urban wastewater treatment process.

Contamination by nanoplastics in urban water has aroused increasing concern. The impact of nanoplastic exposure on the wastewater treatment process in the long term is still unclear. This study investigated the effect of continuous nanoplastic exposure (R1:0, R2:10, R3:100, and R4:1000 µg/L) on the nitrification and denitrification processes for over 200 days in a sequencing batch reactor (SBR). The results revealed that nanoplastic exposure does not demonstrate significant inhibition of total nitrogen removal. The ammonia oxidation rate (19.24 ± 0.01 mgN/gMLVSS/h, p < 0.05) and denitrification rate (11.78 ± 0.11 mgN/ gMLVSS/h, p < 0.05) in R4 was significantly lower than the control (R1: 0 µg/L). The maximal reaction velocities of N2O reduction (Vmax) were improved after long-term exposure to nanoplastics in high concentrations. The R3 demonstrated the highest Vmax value-six times higher than R4 and approximately 20 times higher than R1 and R2. The microbial structure largely varied with the exposure to nanoplastics, where the exposure to a high concentration largely suppressed the nitrifier and selectively enriched the denitrifier. The percentage of the top 20 genera of denitrifiers increased from 31.76% to 63.42%, and the nitrifiers decreased from an initial 12.40% to 2.83% for R4. The predominant genera were found to be Thauera, Azoarcus, and Defluviicoccus in R4 and R3 which indicated their tolerance to nanoplastics. The function prediction results indicated that the membrane transport function was significantly enhanced and the lipid metabolism function was significantly reduced in R4 as compared with the control (R1, p<0.05). This may be attributed to the adsorption of nanoplastics on bacteria. Observation under a scan electronic microscope demonstrated that the nanoplastics were firmly attached to the microbe surface and aggregated in activated sludge at high nanoplastics dosed reactor. These results deepen the understanding of the effect of nanoplastics on the urban wastewater treatment process and provide valuable information for the management of nanoplastic contamination in urban wastewater.

Identification of a key gene StAR-like-3 responsible for carotenoids accumulation in the noble scallop Chlamys nobilis.

Carotenoids play important roles in living organisms. However, animals cannot synthesize carotenoids by themselves, and they must absorb and accumulate carotenoids from their diets in which some key genes are involved. In present study, a gene named StAR-like-3 was characterized in the noble scallop Chlamys nobilis, and its function was identified using golden scallops with higher carotenoids content and brown scallops with less carotenoids content by immunohistochemistry, carotenoid binding assay and RNAi. Results showed that the StAR-like-3 encodes a 54.7 kDa transmembrane protein (named as StAR3) of 481 amino acids containing a MENTAL domain and a START (Steroidogenic acute regulatory protein-related lipid transfer) domain, and its expression level in hemocytes and intestine of golden scallops were significantly higher than those of brown ones. Subsequently, the StAR3 protein was detected in the intestinal epithelial cells of golden scallops, and recombinant StAR3 could bind lutein conjugated to protein G and antibody to form a yellow complex, suggesting it is a carotenoid binding protein involving in carotenoids accumulation in golden scallops. Furthermore, total carotenoids content of hemolymph in golden scallops was significantly decreased when the expression of StAR-like-3 suppressed, suggesting this gene plays an important role in transport of carotenoids. Conclusion, the present results indicated that the StAR-like-3 is a key gene responsible for the carotenoids accumulation in the scallop.

Distinctive Microbial Processes and Controlling Factors Related to Indirect N2O Emission from Agricultural and Urban Rivers in Taihu Watershed.

Inland rivers are hotspots of anthropogenic indirect nitrous oxide (N2O) emissions, but the underlying microbial processes remain poorly understood. This study measured N2O fluxes from agricultural and urban rivers in Taihu watershed and investigated the microbial processes driving N2O production and consumption. The N2O fluxes were significantly higher in agricultural rivers (140.1 ± 89.1 µmol m-2 d-1) than in urban rivers (25.1 ± 27.0 µmol m-2 d-1) (p < 0.001). All wind-based models significantly underestimated N2O flux in urban rivers (p < 0.05) when using the Intergovernmental Panel on Climate Change method because they underestimated the N2O emission factor (EF5r). Wind speed and nitrate were the key factors affecting N2O fluxes in agricultural and urban rivers, respectively. NirK-type denitrifiers produced N2O in urban river water, while nirS-type denitrifiers consumed N2O in the sediments of all rivers. Co-occurrence network analysis indicated organics from Microcystis served as electron donors for denitrifiers (dominated by Flavobacterium) in water, while direct interspecies electron transfer between Thiobacillus and methanogens and between Dechloromonas and sulfate-reducing bacteria enhanced N2O reduction in sediments. This study advances our knowledge on the distinctive microbial processes that determine N2O emissions in inland rivers and illustrates the need to revise EF5r for N2O estimation in urban rivers.

Research on Hot Corrosion Behavior of DZ40M and K452 Superalloys in NaCl Molten Salt.

The corrosion of cobalt-based DZ40M and nickel-based K452 superalloy at 900 °C was investigated by NaCl salt coating. Accordingly, the differences in hot corrosion behavior were analyzed considering the development methods and elementary composition by comparing the two alloys' failure. Then, the corrosion mechanism induced by NaCl was proposed by comparing oxidation and hot corrosion behavior. The relatively continuous Al2O3 and TiO2 formed on K452 superalloy with higher content of Al and Ti have lower solubility and less damage in Na2O. Thus, the hot corrosion rate of K452 is lower than that of DZ40M with higher content of C, Cr, and W.

Fifteen years of the proficiency testing program for HIV-1 viral load testing laboratories in China, 2005-2019.

BACKGROUND: HIV-1 viral load (VL) testing is essential for monitoring the effects of antiretroviral therapy in HIV-infected patients. In order to identify factors that impact testing performance of HIV-1 VL testing laboratories, this study performed a retrospective analysis on data from the domestic HIV-1 VL proficiency testing (PT) program in China during 2005 to 2019. METHODS: Analysis was conducted on testing results of 155 PT panel specimens that were distributed to HIV-1 VL testing laboratories nationwide during 2005 to 2019. Follow-up on-site inspection records on unqualified laboratories were also analyzed. RESULTS: By 2019, 267 HIV-1 VL testing laboratories in China enrolled in the national PT assessment. Overall, HIV-1 VL testing performance has been consistently good after 2012, with less than 5% of participants reporting an unqualified score. Unsatisfactory equipment conditions and poor laboratory administration were the two main reasons causing unqualified scores in the PT assessment. Interestingly, we found that HIV-1 VL testing laboratories had better performance in the CDC system than in hospitals. In analysis on the variance of specimen testing results by different assays, we found that variation in results existed across different assay platforms, and HIV-1 VL testing assays based on real-time PCR technology showed comparatively smaller inter-laboratory variability. CONCLUSIONS: To maintain high performance in HIV-1 VL testing laboratories, it is important to strengthen laboratory management and preserve equipment conditions. Due to the variation of testing results among different assay platforms, we suggest using the same assay platform for longitudinal monitoring of patient viral load.

Physiological and immunological responses to mass mortality in noble scallop Chlamys nobilis cultured in Nan'ao waters of Shantou, China.

The noble scallop Chlamys nobilis has been a commercially important marine cultured bivalve in the Southern Sea of China for decades. Mass mortality events, however, often occur during scallops' cultivation. Mortality of up to 67%-90% was recorded at the beginning of March in 2017 in some culture areas of Nan'ao (Shantou, China), spreading to all scallops within a week. In the present study, in order to investigate the response of the noble scallop at the physiological and molecular level during mass mortality, scallops with different mortalities of 90%, 67%, and 6% were sampled from three sites at Hunter bay, Baisha bay, and Longhai, respectively. Total carotenoids content (TCC), total antioxidant capacity (TAC), malondialdehyde (MDA) content and the expression levels of three immune-related genes (toll-like receptor, C-type lectin receptor and big defensing) in different scallop tissues were determined. The scallops were divided into three groups of sub-health, lesion, and health. TAC, TCC, as well as transcript levels of CnTLR-1, Cnlec-1 and CnBD in sub-health and lesion scallops were all significantly lower (P < 0.05) than those in health scallops, while MDA in sub-health and lesion scallops were significantly higher than those in health scallops (P < 0.05). Similarly, TCC and TAC in lesion scallops were both higher than sub-health scallops. Moreover, significantly positive correlations were found between TCC and TAC (P < 0.05) and between CnTLR-1 and Cnlec-1 (P < 0.05), while significantly negative correlations were found between TCC and CnTLR-1 (P < 0.05), TCC and Cnlec-1 (P < 0.05), TAC and CnBD (P < 0.05), as well as between MDA and Cnlec-1 (P < 0.001). All the results indicate that noble scallops significantly change their physiological and molecular levels when suffering from stress, and that their antioxidant and immune response systems play important defense functions.

Sodium fluoride induces apoptosis in odontoblasts via a JNK-dependent mechanism.

Sodium fluoride (NaF) is widely used for the treatment of dental caries and dentin hypersensitivity. However, its pro-apoptotic effect on odontoblasts may lead to harmful side-effects. The purpose of this study was to evaluate the pro-apoptotic effects of NaF in odontoblasts and elucidate the possible underlying molecular mechanisms. NaF generated cytotoxic effects in odontoblast-lineage cell (OLC) in a dose- and time-dependent manner. Exposure of cells to 4mM NaF for 24h induced caspase-3 activation, ultrastructural alterations, and resulted in the translocation of Bax to the mitochondria and the release of cytochrome c from the mitochondrial inter-membrane space into the cytosol, indicating that fluoride-mediated apoptosis is mitochondria-dependent. Fluoride treatment also increased phosphorylation of JNK and ERK, but not p38, and apoptosis induced by fluoride was notably or partly suppressed by treatment with JNK or ERK inhibitors, respectively. Taken together, these findings suggest that NaF induces apoptosis in OLC odontoblasts through a JNK-dependent mitochondrial pathway.

Antimicrobial and antibiofilm activity of pleurocidin against cariogenic microorganisms.

Dental caries is a common oral bacterial infectious disease of global concern. Prevention and treatment of caries requires control of the dental plaque formed by pathogens such as Streptococcus mutans and Streptococcus sobrinus. Pleurocidin, produced by Pleuronectes americanus, is an antimicrobial peptide that exerts broad-spectrum activity against pathogenic bacteria and fungi. Moreover, pleurocidin shows less hemolysis and is less toxic than other natural peptides. In the present study, we investigated whether pleurocidin is an effective antibiotic peptide against common cariogenic microorganisms and performed a preliminary study of the antimicrobial mechanism. We assayed minimal inhibitory concentration (MIC), minimal bactericide concentration (MBC) and bactericidal kinetics and performed a spot-on-lawn assay. The BioFlux system was used to generate bacterial biofilms under controllable flow. Fluorescence microscopy and confocal laser scanning microscopy (CLSM) were used to analyze and observe biofilms. Scanning electron microscopy was used to observe the bacterial membrane. MIC and MBC results showed that pleurocidin had different antimicrobial activities against the tested oral strains. Although components of saliva could affect antimicrobial activity, pleurocidin dissolved in saliva still showed antimicrobial effects against oral microorganisms. Furthermore, pleurocidin showed a favorable killing effect against BioFlux flow biofilms in vitro. Our findings suggest that pleurocidin has the potential to kill dental biofilms and prevent dental caries.