Effects of two strains of thermophilic nitrogen-fixing bacteria on nitrogen loss mitigation in cow dung compost.

Excavating nitrogen-fixing bacteria with high-temperature tolerance is essential for the efficient composting of animal dung. In this study, two strains of thermophilic nitrogen-fixing bacteria, NF1 (Bacillus subtilis) and NF2 (Azotobacter chroococcum), were added to cow dung compost both individually (NF1, NF2) and mixed together (NF3; mixing NF1 and NF2 at a ratio of 1:1). The results showed that NF1, NF2, and NF3 inoculants increased the total Kjeldahl nitrogen level by 38.43%-55.35%, prolonged the thermophilic period by 1-13 d, increased the seed germination index by 17.81%, and the emissions of NH3 and N2O were reduced by 25.11% and 42.75%, respectively. Microbial analysis showed that Firmicutes were the predominant bacteria at the thermophilic stage, whereas Chloroflexi, Proteobacteria, and Bacteroidetes were the predominant bacteria at the mature stage. These results confirmed that the addition of the isolated strains to cow dung composting improved the bacterial community structure and benefited nitrogen retention.

Mechanisms and effects of novel ammonifying microorganisms on nitrogen ammonification in cow manure waste composting.

It is essential to reduce nitrogen losses and to improve nitrogen conversion during organic waste composting because of environmental protection and sustainable development. To reveal newly domesticated ammonifying microorganisms (AM) cultures on the ammonification and nitrogen conversion during the composting, the screened microbial agents were inoculated at 5 % concentration (in weight basis) into cow manure compost under five different treatments: sterilized distilled water (Control), Amm-1 (mesophilic fungus-F1), Amm-2 (mesophilic bacterium-Z1), Amm-3 (thermotolerant bacterium-Z2), and Amm-4 (consortium: F1, Z1, and Z2), and composted for 42 days. Compared to control, AM inoculation prolonged the thermophilic phases to 9-19 days, increased the content of NH4+-N to 1.60-1.96 g/kg in the thermophilic phase, reduced N2O and NH3 emissions by 22.85-61.13 % and 8.45-23.29 %, increased total Kjeldahl nitrogen, and improved cell count and viability by 12.09-71.33 % and 66.71-72.91 %. AM was significantly associated with different nitrogen and microbial compositions. The structural equation model (SEM) reveals NH4+-N is the preferable nitrogen for the majority of bacterial and fungal growth and that AM is closely associated with the conversion between NH3 and NH4+-N. Among the treatments, inoculation with Amm-4 was more effective, as it significantly enhanced the driving effect of the critical microbial composition on nitrogen conversion and accelerated nitrogen ammonification and sequestration. This study provided new concepts for the dynamics of microbial in the ammonification process of new AM bacterial agents in cow manure compost, and an understanding of the ecological mechanism underlying the ammonification process and its contribution to nitrogen (N) cycling from the perspective of microbial communities.

Diluent pH affects sperm motility via GSK3 α/ß-hexokinase pathway for the efficient enrichment of X-sperm to increase the female kids rate of dairy goats.

Dairy goats are the goats bred with the ability to produce large quantities of milk, and the increase of the female kid rate of breeding dairy goats is beneficial for milk production and economic benefits of dairy goat farms. Our previous study revealed that regulating the pH of dairy goat semen diluent to 6.2 or 7.4 respectively, the proportion of X chromosome bearing sperm (X-sperm) in the up and down layers of the tube after incubation was significantly higher than that of Y chromosome bearing sperm (Y-sperm) i.e. enriched X-sperm. In this study, fresh dairy goat semen collected in different seasons was diluted in different pH solutions to calculate the number and rate of X-sperm and to measure the functional parameters of enriched sperm. The artificial insemination experiments were performed with enriched X-sperm. The mechanisms of regulating the pH of diluent affecting sperm enrichment were further studied. The results showed that the proportion of enriched X-sperm in pH 6.2 and 7.4 diluents of sperm collected in different seasons showed no significantly different, but were significantly higher than that of the control group (pH 6.8). The in vitro functional parameters of X-sperm enriched in pH 6.2 and 7.4 diluent solution were not significantly different from those of the control group (P > 0.05). After artificial insemination with X-sperm enriched in pH7.4 diluent, the proportion of female offspring was significantly higher than that of the control group. It was found that the regulating pH of the diluent affected sperm mitochondrial activity and glucose uptake capacity via phosphorylating NF-κB and GSK3α/ß proteins. The motility activity of X-sperm was enhanced under acidic conditions and weakened under alkaline conditions, which was conducive to the effective enrichment of X-sperm. This study demonstrated that the number and proportion of X-sperm enriched using pH 7.4 diluent were elevated, and the proportion of female kids was increased. This technology can be used for the reproduction and production of dairy goats in farms at large scales.

Alkaline Dilution Alters Sperm Motility in Dairy Goat by Affecting sAC/cAMP/PKA Pathway Activity.

In dairy goat farming, increasing the female kid rate is beneficial to milk production and is, therefore, economically beneficial to farms. Our previous study demonstrated that alkaline incubation enriched the concentration of X-chromosome-bearing sperm; however, the mechanism by which pH affects the motility of X-chromosome-bearing sperm remains unclear. In this study, we explored this mechanism by incubating dairy goat sperm in alkaline dilutions, examining the pattern of changes in sperm internal pH and Ca2+ concentrations and investigating the role of the sAC/cAMP/PKA pathway in influencing sperm motility. The results showed that adding a calcium channel inhibitor during incubation resulted in a concentration-dependent decrease in the proportion of spermatozoa with forward motility, and the sperm sAC protein activity was positively correlated with the calcium ion concentration (r = 0.9972). The total motility activity, proportion of forward motility, and proportion of X-chromosome-bearing sperm decreased (p < 0.05) when cAMP/PKA protease activity was inhibited. Meanwhile, the enrichment of X-chromosome-bearing sperm by pH did not affect the sperm capacitation state. These results indicate that alkaline dilution incubation reduces Ca2+ entry into X-sperm and the motility was slowed down through the sAC/cAMP/PKA signaling pathway, providing a theoretical foundation for further optimization of the sex control method.