Efficient inactivation of amoeba spores and their intraspore bacteria by solar/chlorine: Kinetics and mechanisms.

Amoebae are widespread in water and serve as environment vectors for pathogens, which may threaten public health. This study evaluated the inactivation of amoeba spores and their intraspore bacteria by solar/chlorine. Dictyostelium discoideum and Burkholderia agricolaris B1qs70 were selected as model amoebae and intraspore bacteria, respectively. Compared to solar irradiation and chlorine, solar/chlorine enhanced the inactivation of amoeba spores and intraspore bacteria, with 5.1 and 5.2-log reduction at 20 min, respectively. The enhancement was similar in real drinking water by solar/chlorine under natural sunlight. However, the spore inactivation decreased to 2.97-log by 20 min solar/chlorine under oxygen-free condition, indicating that ozone played a crucial role in the spore inactivation, as also confirmed by the scavenging test using tert­butanol to scavenge the ground-state atomic oxygen (O(3P)) as a ozone precursor. Moreover, solar/chlorine induced the shape destruction and structural collapse of amoeba spores by scanning electron microscopy. As for intraspore bacteria, their inactivation was likely ascribed to endogenous reactive oxygen species. As pH increased from 5.0 to 9.0, the inactivation of amoeba spores decreased, whereas that of intraspore bacteria was similar at pH 5.0 and 6.5 during solar/chlorine treatment. This study first reports the efficient inactivation of amoeba spores and their intraspore pathogenic bacteria by solar/chlorine in drinking water.

Protozoa as Hotspots for Potential Pathogens in the Drinking Water of a Subtropical Megacity: Diversity, Treatment, and Health Risk.

Drinking water systems host a wide range of microorganisms essential for biosafety. However, one major group of waterborne pathogens, protozoa, is relatively neglected compared to bacteria and other microorganisms. Until now, little is known about the growth and fate of protozoa and their associated bacteria in drinking water systems. In this study, we aim to investigate how drinking water treatment affects the growth and fate of protozoa and their associated bacteria in a subtropical megacity. The results showed that viable protozoa were prevalent in the city's tap water, and amoebae were the major component of tap water protozoa. In addition, protozoan-associated bacteria contained many potential pathogens and were primarily enriched in amoeba hosts. Furthermore, this study showed that current drinking water disinfection methods have little effect on protozoa and their associated bacteria. Besides, ultrafiltration membranes unexpectedly served as an ideal growth surface for amoebae in drinking water systems, and they could significantly promote the growth of amoeba-associated bacteria. In conclusion, this study shows that viable protozoa and their associated bacteria are prevalent in tap water, which may present an emerging health risk in drinking water biosafety.

Environmental stress promotes the persistence of facultative bacterial symbionts in amoebae.

Amoebae are one major group of protists that are widely found in natural and engineered environments. They are a significant threat to human health not only because many of them are pathogenic but also due to their unique role as an environmental shelter for pathogens. However, one unsolved issue in the amoeba-bacteria relationship is why so many bacteria live within amoeba hosts while they can also live independently in the environments. By using a facultative amoeba- Paraburkholderia bacteria system, this study shows that facultative bacteria have higher survival rates within amoebae under various environmental stressors. In addition, bacteria survive longer within the amoeba spore than in free living. This study demonstrates that environmental stress can promote the persistence of facultative bacterial symbionts in amoebae. Furthermore, environmental stress may potentially select and produce more amoeba-resisting bacteria, which may increase the biosafety risk related to amoebae and their intracellular bacteria.

Enhanced microbial nitrification-denitrification processes in a subtropical metropolitan river network.

Urban rivers are hotspots of regional nitrogen (N) pollution and N transformations. Previous studies have reported that the microbial community of urban rivers was different from that of natural rivers. However, how microbial community affects N transformations in the urban rivers is still unclear. In this study, we employed N nutrients-related isotope technology (includes natural-abundance isotopes survey and isotope-labeling method) and bioinformatics methods (includes 16S rRNA high-throughput sequencing and quantitative PCR analysis) to investigate the major N transformations, microbial communities as well as functional gene abundances in a metropolitan river network. Our results suggested that the bacterial community structure in the highly urbanized rivers was characterized by higher richness, less complexity and increased abundances of nitrification and denitrifying bacterium compared to those in the suburban rivers. These differences were mainly caused by high sewage discharge and N loadings. In addition, the abundances of nitrifier gene (amoA) and denitrifier genes (nirK and nirS) were significantly higher in the highly urbanized rivers (2.36 × 103, 7.43 × 107 and 2.28 × 107 copies·mL-1) than that in the suburban rivers (0.43 × 103, 2.18 × 107 and 0.99 × 107 copies·mL-1). These changes in microbes have accelerated nitrification-denitrification processes in the highly urbanized rivers as compared to those in the suburban rivers, which was evidenced by environmental isotopes and the rates of nitrification (10.52 vs. 0.03 nmol·L-1·h-1) and denitrification (83.31 vs. 22.49 nmol·g-1·h-1). Overall, this study concluded that the excess exogenous N has significantly shaped the specific aquatic bacterial communities, which had a potential for enhancing nitrification-denitrification processes in the highly urbanized river network. This study provides a further understanding of microbial N cycling in urban river ecosystems and expands the combined application of isotopic technology and bioinformatics methods in studying biogeochemical cycling.

Complex Bilateral Interactions Determine the Fate of Polystyrene Micro- and Nanoplastics and Soil Protists: Implications from a Soil Amoeba.

Nano- and microplastics have become a serious global concern, threatening our living environments. Previous studies have shown that many organisms, including bacteria, animals, and plants, can be affected by microplastics. However, little is known about one ecologically important group of soil organisms, the protists. In this study, we investigated how polystyrene micro- and nanoplastics interacted with a soil amoeba Dictyostelium discoideum. The results showed that environmental concentrations of nano- and microplastics could negatively affect the soil amoeba's fitness and development. D. discoideum ingested both nano- and microplastics through phagocytosis but packed and excreted them during slug migration, which also promoted their biodegradation. Fourier transform infrared spectroscopy analyses revealed the formation of new oxygen-containing functional groups and the sign of possible oxidation of polystyrene. Also, nano- and microplastic exposure disrupted the nutrient and energy metabolisms of D. discoideum and affected the expression of key genes (e.g., cf45-1, dcsA, aprA, dymB, and gefB) related to morphogenesis and phagocytosis. In conclusion, our results show that nano- and microplastics have complex bilateral interactions with the soil amoeba, affecting each other's fate in the soil environment. This study provides new insights into how soil protists interact with nano- and microplastics in the soil ecosystem.

Coupling oxidation of acid volatile sulfide, ferrous iron, and ammonia nitrogen from black-odorous sediment via autotrophic denitrification-anammox by nitrate addition.

The coupling removal of acid volatile sulfide (AVS), ferrous iron, and ammonia nitrogen has been applied for black-odorous sediment remediation. In this study, calcium nitrate with different N/(S + Fe) ratios (0.45, 0.90, 1.20 and 1.80) was added into black-odorous sediment in four systems named R1, R2, R3, and R4. Results showed that the removal rate of AVS was 76.40% in the R1, which was lower compared with rates in R2-R4 around 96.70%. The ferrous oxidation rate was approximately 87.00% in R2-R4, which was considerably higher than that in the R1 (24.62%). And the ammonia was reduced by 81.02%, 88.00%, 100%, and 57.18% in R1, R2, R3 and R4, respectively. During the reaction, nitrite accumulation was observed, indicating partial denitrification. Moreover, microbes related to autotrophic denitrification (e.g., genus Thiobacillus, Dok59, GOUTA19, Gallionella, with the highest abundance of 15.40%, 13.21%, 8.79%, 9.44%, respectively) were detected in all systems. Furthermore, the anammox bacteria Candidatus_Brocadia with the highest abundance of 3.44% and 4.00% in R2 and R3, respectively was also found. These findings confirmed that AVS, ferrous iron, and ammonia nitrogen could be simultaneously removed via autotrophic denitrification coupled with anammox in black-odorous sediment by nitrate addition.