Probiotics improve polystyrene microplastics-induced male reproductive toxicity in mice by alleviating inflammatory response.

As a new type of environmental pollutant, microplastics have been garnered increasing attention, especially in regard to their effects on the reproductive system. However, researchers have yet to report whether prevention and treatment measures exist for reproductive injury caused by microplastics. The aim of this study was therefore to explore the mechanism of spermatogenic injury induced by polystyrene microplastics (PS-MPs) and the intervention effect of probiotics based on the gut microbiota-testis axis. Mice were orally exposed for 35 days to 5 µm of PS-MPs with a gavage dose was 0.1 mg/day, and the intervention group was given probiotics (Lactobacillus, Bifidobacterium longum, and Enterococcus) orally. Fecal samples were then subjected to 16 S rRNA sequencing analysis, and sperm motion was analyzed by a Hamilton-Thorne Sperm analyzer. The results showed that PS-MPs exposed mice had significant spermatogenic dysfunction and testicular inflammation. In addition, the intestinal microbial structure of exposed mice changed significantly; the abundance of Lactobacillus decreased, and the abundance of Prevotella increased. Furthermore, with fecal microbiota transplantation, the recipient mice showed a significant decrease in sperm quality. However, probiotics supplementation helped inhibit the activation of IL-17A signaling driven by gut microbes, thereby alleviating the inflammatory response and improving sperm quality decline caused by PS-MPs. These results may provide a scientific basis for further understanding of the mechanism of male reproductive damage caused by environmental pollutants such as microplastics and for novel reproductive damage intervention measures.

Polystyrene microplastics induce mitochondrial damage in mouse GC-2 cells.

Microplastics are widely distributed, such as oceans, rivers and the atmosphere, with many opportunities for human exposure and potential health risks. Polystyrene microplastic (PS-MPS) exposure has been found to cause sperm damage to mice; however, the mechanism by which this happens remains unclear. Here, GC-2 cells, a mouse spermatocyte line, were exposed to 5 µm PS-MPS to investigate mitochondrial damage. The results showed that 5 µm PS-MPS decreased ATP content, reduced the mitochondrial membrane potential, damaged the integrity of the mitochondrial genome, and caused an imbalance of homoeostasis between mitochondrial division and fusion. The mitochondrial PINK1/Parkin autophagy pathway was activated. Time-series analysis revealed that PS-MPS damaged the mitochondrial structure through cellular oxidative stress, and mitochondrial function was maintained to some extent after PS-MPS damage. This study revealed the mitochondrial toxicity of polystyrene microplastics, thus providing a basis for understanding the causes of sperm damage by polystyrene microplastics.

Synergy of N-(3-oxohexanoyl)-L-homoserine lactone and tryptophan-like outer extracellular substances in granular sludge dominated by aerobic ammonia-oxidizing bacteria.

Nitrogen removal via nitrite is an energy-saving method for high-strength ammonia wastewater treatment. A better understanding of the formation of granular sludge dominated by aerobic ammonia-oxidizing bacteria (AerAOB) could facilitate the improved use of rapid sludge granulation for nitritation. In this study, AerAOB-dominated activated sludge (NAS) and granular sludge (NGS) produced different N-scyl-homoserine lactones (AHLs). N-(3-oxohexanoyl)-L-homoserinelactone (OHHL), only released from NGS, was shown to accelerate sludge aggregation by increasing the biomass growth rate, microbial activity, extracellular protein, and AerAOB biomass. For both NAS and NGS, sludge cells were glued together by inner extracellular polymeric substances (EPSs) with similar components to form microcolony. Different from the characterized negative effect of NAS's outer-EPS on cell adhesion, the outer-EPS of NGS played a positive role in the attached growth of AerAOB-dominated sludge and contained more tryptophan-like substances. More interesting, OHHL enhanced the yields of tryptophan-like substances after mixing with the outer-EPS of NGS, enhancing cell adhesion. In a word, OHHL and more tryptophan-like substances were produced in the process of granulation under the selective sludge discharge condition, which was proved to be able to accelerate NAS granulation. Therefore, the sludge granulation process for nitritation can be improved by increasing the levels of OHHL and tryptophan in the initial startup stage. The appropriate engineering strategy should be further studied to facilitate the actual application of granular sludge for nitrogen removal on a large scale.

Inhibition of NF-κB activation improves insulin resistance of L6 cells.

To explore the role of NF-κB activation in the development of insulin resistance and investigate whether or not that the inhibition of NF-κB activation by PDTC will improve the insulin resistance of L6 cells exposed to H2O2. L6 cells were treated with H2O2, PDTC or both H2O2 and PDTC for 4 hours. The uptake of glucose with stimulation of insulin, the expression of P38-MAPK, p- P38-MAPK, NF-κBp65, p- NF-κBp65, IRS-1, IRS-2, p-IRS-2, PI3K, IκBα, p- IκBα, caspase-8 and GLUT4, the production of ROS, TNF-α, IL-6 and IL-1ß as well as the apoptosis rate of L6 cells were determined and compared in L6 treated with H2O2 alone or both H2O2 and PDTC. Compared with the L6 cells treated with H2O2 alone, the L6 cells treated with both H2O2 and PDTC showed (1) significantly lower production of ROS, TNF-α, IL-6 and IL-1ß; (2) significantly decreased expression of P38-MAPK, p- P38-MAPK and NF-κBp65, p- NF-κBp65, p- IκBα and caspase-8; (3) significantly lower rate of apoptosis; (4) significantly higher expression of IRS-2, p-IRS-2 (Tyr 612), PI3K and GLUT4; (5) significantly higher uptake of glucose with stimulation of insulin; (6) significantly increased expression of Bcl2 and decreased ratio of Bax to Bcl2. Based on the findings of the present study, inhibition of NF-κB activation by PDTC would improve the insulin resistance of L6 cells exposed to H2O2.

Determination of Biological and Molecular Attributes Related to Polystyrene Microplastic-Induced Reproductive Toxicity and Its Reversibility in Male Mice.

Microplastics exist not only in the natural environment, but also in human tissue such as blood and even placenta. Polystyrene microplastic exposure can cause abnormal sperm quality in mice; however, the mechanism is unclear, and whether sperm abnormalities can be restored has not been reported. ICR mice were exposed to 5 µm polystyrene microplastics through the drinking water. After one spermatogenic cycle, mitochondrial damage was observed to explain the possible cause of sperm damage. After 1-2 spermatogenic cycles of recovery, whether the damaged sperm could be recovered was observed. The results show that polystyrene microplastics caused a decrease in the mitochondrial membrane potential, an imbalance of kinetic homeostasis, a change in genetic characteristics, mitophagy, and a decrease in the ATP content in mouse testicular tissue. Oxidative stress may be the cause of mitochondrial damage. After 1-2 spermatogenic cycles, mitochondrial damage was restored and sperm quality was improved. This study explored the mitochondrial causes of reproductive toxicity of polystyrene microplastics and the reversibility of reproductive toxicity, providing data for further research on the toxicity of microplastics and the prevention and treatment of its harm.

Reproductive toxicity of polystyrene microplastics: In vivo experimental study on testicular toxicity in mice.

Microplastics (MPS) are widespread in our environment and have a potential impact on the reproductive development of humans and mammals. In this study, we evaluated the effect of 5 µm polystyrene microplastics(PS-MPS) on spermatogenesis in mice. The damage by PS-MPS to epididymal sperm was studied using blood cell counts. The results showed that the number of viable epididymis sperm after PS-MPS exposure was significantly reduced. Using Duff-Quik staining, we found that the PS-MPS exposure increased the rate of sperm deformity. The testis is an important organ responsible for normal spermatogenesis. HE and TUNEL staining showed atrophy, shedding, and apoptosis of sperm cells at all levels of the testis after exposure to PS-MPS. Western blot and qPCR analysis were used to detect Nrf2/HO-1 and NF-κB. The results showed that after PS-MPS exposure, the expression of the pro-inflammatory molecule NF-κB and that of the inflammatory factors interleukin (IL)-1ß and IL-6 increased significantly, whereas that of the anti-inflammatory molecule Nrf2/HO-1 decreased. These results indicate that the abnormal sperm quality in ICR mice caused by PS-MPS exposure is closely related to the Nrf2/HO-1/NF-κB pathway.

Cell adhesion, ammonia removal and granulation of autotrophic nitrifying sludge facilitated by N-acyl-homoserine lactones.

In this study, six N-acyl-homoserine lactone (AHL) molecules (C6-HSL, C8-HSL, C10-HSL, 3-oxo-C6-HSL, 3-oxo-C8-HSL and 3-oxo-C10-HSL) were each dosed into a bioreactor and seeded using autotrophic nitrifying sludge (ANS). The effects of the AHLs on cell adhesion, nitrification and sludge granulation were investigated. The results indicated that the efficiencies of cell adhesion and ammonia removal both had a close correlation with the side chain length and ß position substituent group of the AHLs. The best-performing AHL in terms of accelerating bacterial attached-growth was 3-oxo-C6-HSL, whereas C6-HSL outperformed the others in terms of the ammonia degradation rate. The addition of 3-oxo-C6-HSL or C6-HSL increased the biomass growth rate, microbial activity, extracellular proteins and nitrifying bacteria, which can accelerate the formation of nitrifying granules. Consequently, selecting AHL molecules that could improve bacteria in attached-growth mode and nitrification efficiency simultaneously will most likely facilitate the rapid granulation of nitrifying sludge.