[Effect of chromosomal polymorphisms on reproductive outcomes of in vitro fertilization-embryo transfer].

OBJECTIVE: To explore the effect of chromosomal polymorphic variations on the outcome of in vitro fertilization/intracytoplasmic sperm injection-embryo transfer (IVF/ICSI-ET) treatment for infertile couples. Methods： In this retrospective study, 418 infertile couples, who received their first IVF/ICSI-ET treatment cycles in Reproductive Medicine Center of Xi'an People's Hospital (Xi'an Fourth Hospital) from Jan. 2021to Jan. 2023, were included and divided into two groups: group A (the infertile couples with normal chromosome) and group B(with chromosomal polymorphic variations); The group B divided into 2 groups, group C(male carrier, n= 44), group D (female carrier, n= 37). The No. of oocyte, 2PN fertilization rate, fertilization rate,multi-PN fertilization rate, quality embryo rate, cleavage rate, normal cleavage rate, blastocyst formation rate,clinical pregnancy rate after IVF/ICSI-ET treatment were compared and analyzed. Logistic regression analysis was performed to study the relationship between clinical pregnancy rate and chromosome variation. Results： A total of 418 infertile couples received IVF/ICSI-ET treatment were enrolled, with 81 in the chromosomal polymorphic group (group A), and 337 in the normal chromosomal group (group B). The quality embryo rate were signifcantly higher in the group A（59.21% vs 52.42%,P<0.05）,the other outcomes have no significant differences between the two groups ( P>0.05). The No. of oocyte, 2PN fertilization rate, fertilization rate,multi-PN fertilization rate, quality embryo rate, cleavage rate, normal cleavage rate, blastocyst formation rate showed no signifcantdiferences among group A, group C and group D (P>0.05). The clinical pregnancy rate in group A is higer than group C and group D (56.7% vs 52.3% vs 40.5%), But there were no signifcantly diferent among the three groups (P>0.05). Logistic regression analysis indicated that the Chromosomal polymorphisms were not associated with clinical pregnancy rate (P>0.05).Conclusion： Chromosomal polymorphisms appear to have no significant effect on the outcome of IVF/ICSI-ET treatment for infertile couples.

Stress-responses of microbes in oil reservoir under high tetracycline exposure and their environmental risks.

As the groundwater ecosystem is connected with surface, antibiotics and antibiotic resistance genes (ARGs) in aquatic environments will gradually infiltrate into the deep environment, posing a potential threat to groundwater ecosystem. However, knowledge on the environmental risk of antibiotics and ARGs in groundwater ecosystem and their ecological process still remains unexplored. In this study, lab-scale oil reservoirs under high tetracycline stress were performed to evaluate the dynamics of microbial communities, ARGs and potential functions by using 16S rRNA gene sequencing and metagenomics analysis. Although the presence of antibiotics remarkably reduced the microbial abundance and diversity in a short term, but remain stable or even increased after a long-term incubation. Antibiotic stress caused a greater diversity and abundance of ARGs, and higher numbers of ARGs-related species with the capacity to transfer ARGs to other microbes through horizontal gene transfer. Thus, a much more frequent associations of microbial community at both node- and network-level and a selective pressure on enrichment of antibiotic resistant bacteria related to "anaerobic n-alkane degradation" and "methylotrophic methanogenesis" were observed. It is important to emphasize that high antibiotic stress could also prevent some microbes related to "Sulfate reduction", "Fe(II) oxidation", "Nitrate reduction", and "Xylene and Toluene degradation". This study provides an insight into the long-term stress-responses of microbial communities and functions in oil reservoir under tetracycline exposure, which may help to elucidate the effect of antibiotic stress on biogeochemical cycling with microbial involvement in groundwater ecosystem.

Deep mining decreases the microbial taxonomic and functional diversity of subsurface oil reservoirs.

Microbes in subsurface oil reservoirs play important roles in elemental cycles and biogeochemical processes. However, the community assembly pattern of indigenous microbiome and their succession under long-term human activity remain poorly understood. Here we studied the microbial community assembly in underground sandstone cores from 190 to 2050 m in northeast China and their response to long-term oil recovery (10-50 years). Indigenous microbiome in subsurface petroleum reservoirs were dominated by Gammaproteobacteria, Firmicutes, Alphaproteobacteria, Bacteroidetes, and Actinobacteria, which exhibited a higher contribution of homogenizing dispersal assembly and different taxonomy distinct ecological modules when compared with perturbed samples. Specifically, the long-term oil recovery reduced the bacterial taxonomic- and functional-diversity, and increased the community co-occurrence associations in subsurface oil reservoirs. Moreover, distinguished from the perturbed samples, both variation partition analysis and structural equation model revealed that the contents of quartz, NO3- and Cl- significantly structured the α- and ß-diversity in indigenous subsurface bacterial communities. These findings first provide the holistic picture of microbiome in the deep oil reservoirs, which demonstrate the significant impact of human activity on microbiome in deep continental subsurface.

Stochastic assembly process dominates bacterial succession during a long-term microbial enhanced oil recovery.

Microbial enhanced oil recovery (MEOR) has been successfully used in oil exploitation to increase oil production. However, the mechanisms of microbial interactions and community assembly related to oil production performance along MEOR process are poorly understood. Here, we investigated the microbiome of an oil reservoir for a period of 5 years under three phases of different treatments with the injection of a mixture of microbes, nutrients, and air at different intensity. During the MEOR process, amplification of functional genes revealed an increase of genes related to hydrocarbon degradation linked to methanogenesis, supported by stable isotope analysis for confirmation of the methanogenesis activity. Meanwhile, a lower contribution of the ubiquitous/common taxa, closer and more positive associations, and lower modularity were observed in bacterial co-occurrence networks, with the rare taxa being the keystone taxa. The null model analysis and structural equation modeling revealed that the contribution of stochastic processes affected by functional groups and co-occurrence patterns to bacterial community increased significantly with the increase of oil production. This provides new insight that stochastic assembly in bacterial community increased along with MEOR process, and it is worthwhile paying attention to the uncertain consequences caused by random evolution since the treatment effect of MEOR is closely related to the in-situ community in oil reservoir.

Occurrence of antibiotic resistance genes in an oilfield's water re-injection systems.

The recent widespread increase in antibiotic resistance has become a real threat to both human and environmental ecosystem health. In oil reservoirs, an extreme environment potentially influenced by human activity such as water flooding, the distribution and abundance of antibiotic resistance genes (ARGs) remains poorly understood. Herein, we investigated the distribution of ARGs at different positions in a water-flooding oilfield in China, and found that ARGs were observed in all parts of the investigated system. The surface regions of the water re-injection system were more vulnerable to ARG pollution, and the final ARG concentration was up to 2.2 × 108 gene copies/L, and sulfonamide were the most abundant. However, ARG concentration decreased sharply in the samples from underground part of the re-injection system. The bacterial community composition was also varied with sampling position. The sample from production well, which was enriched in crude oil, contained more bacteria but the community richness was simpler. This study also indicated the wastewater-recycling process above ground, which proposed to reduce the discharge into environment directly, may pose a risk for ARGs spread.

Spatial isolation and environmental factors drive distinct bacterial and archaeal communities in different types of petroleum reservoirs in China.

To investigate the spatial distribution of microbial communities and their drivers in petroleum reservoir environments, we performed pyrosequencing of microbial partial 16S rRNA, derived from 20 geographically separated water-flooding reservoirs, and two reservoirs that had not been flooded, in China. The results indicated that distinct underground microbial communities inhabited the different reservoirs. Compared with the bacteria, archaeal alpha-diversity was not strongly correlated with the environmental variables. The variation of the bacterial and archaeal community compositions was affected synthetically, by the mining patterns, spatial isolation, reservoir temperature, salinity and pH of the formation brine. The environmental factors explained 64.22% and 78.26% of the total variance for the bacterial and archaeal communities, respectively. Despite the diverse community compositions, shared populations (48 bacterial and 18 archaeal genera) were found and were dominant in most of the oilfields. Potential indigenous microorganisms, including Carboxydibrachium, Thermosinus, and Neptunomonas, were only detected in a reservoir that had not been flooded with water. This study indicates that: 1) the environmental variation drives distinct microbial communities in different reservoirs; 2) compared with the archaea, the bacterial communities were highly heterogeneous within and among the reservoirs; and 3) despite the community variation, some microorganisms are dominant in multiple petroleum reservoirs.

Comparison of ascorbate metabolism in fruits of two citrus species with obvious difference in ascorbate content in pulp.

Citrus fruit is widely consumed and provides ascorbate for human health. The ascorbate content in pulp is generally higher in orange (Citrus sinensis Osb.) than in Satsuma mandarin (Citrus unshiu Marc.). However, what contributes to such difference is still unknown. In the present study, ascorbate accumulation, expression profiles of genes involved in L-galactose pathway and activity changes of enzymes related with L-ascorbic acid (AA) oxidation and recycling were investigated during fruit development and ripening in fruit pulp of Satsuma mandarin and orange. As fruit ripens, total ascorbate (T-ASC) or AA content increased in mandarin whereas fluctuated on a relatively high level in orange. Concentrations of T-ASC or AA in pulp of orange were over 1.5-fold higher than that in pulp of Satsuma mandarin during fruit ripening. Further analysis showed that each transcript of four genes (encoding GDP-D-mannose-3',5'-epimerase, GDP-L-galactose-pyrophosphatase, L-galactose dehydrogenase and L-galactono-1,4-lactone dehydrogenase respectively) in orange was almost on a higher level and the activities of oxidation enzymes (ascorbate oxidase and ascorbate peroxidase) were lower during fruit ripening as compared with Satsuma mandarin. As ascorbate pool size is decided by the combination of biosynthesis, oxidation and recycling, therefore, higher expression of four genes along with lower activity of oxidation enzymes should contribute at least partially to the higher ASC accumulation in orange pulp.