Bacterial isolation and genome analysis of a novel Klebsiella quasipneumoniae phage in southwest China's karst area.

BACKGROUND: Southwest China is one of the largest karst regions in the world. Karst environment is relatively fragile and vulnerable to human activities. Due to the discharge of sewage and domestic garbage, the karst system may be polluted by pathogenic bacteria. The detection of bacterial distribution and identification of phage capable of infecting them is an important approach for environmental assessment and resource acquisition. METHODS: Bacteria and phages were isolated from karst water in southwest China using the plate scribing and double plate method, respectively. Isolated phage was defined by transmission electron microscopy, one-step growth curve and optimal multiplicity of infection (MOI). Genomic sequencing, phylogenetic analysis, comparative genomic and proteomic analysis were performed. RESULTS: A Klebsiella quasipneumoniae phage was isolated from 32 isolates and named KL01. KL01 is morphologically identified as Caudoviricetes with an optimal MOI of 0.1, an incubation period of 10 min, and a lysis period of 60 min. The genome length of KL01 is about 45 kb, the GC content is 42.5%, and it contains 59 open reading frames. The highest average nucleotide similarity between KL01 and a known Klebsiella phage 6939 was 83.04%. CONCLUSIONS: KL01 is a novel phage, belonging to the Autophagoviridae, which has strong lytic ability. This study indicates that there were not only some potential potentially pathogenic bacteria in the karst environment, but also phage resources for exploration and application.

Microbial difference and its influencing factors in ice-covered lakes on the three poles.

Most lakes in the world are permanently or seasonally covered with ice. However, little is known about the distribution of microbes and their influencing factors in ice-covered lakes worldwide. Here we analyzed the microbial community composition in the waters of 14 ice-covered lakes in the Hoh Xil region of northern Qing-Tibetan Plateau (QTP), and conducted a meta-analysis by integrating published microbial community data of ice-covered lakes in the tripolar regions (the Arctic, Antarctica and QTP). The results showed that there were significant differences in microbial diversity, community composition and distribution patterns in the ice-covered tripolar lakes. Microbial diversity and richness were lower in the ice-covered QTP lakes (including the studied lakes in the Hoh Xil region) than those in the Arctic and Antarctica. In the ice-covered lakes of Hoh Xil, prokaryotes are mainly involved in S-metabolic processes, making them more adaptable to extreme environmental conditions. In contrast, prokaryotes in the ice-covered lakes of the Arctic and Antarctica were predominantly involved in carbon/nitrogen metabolic processes. Deterministic (salinity and nutrients) and stochastic processes (dispersal limitation, homogenizing dispersal and drift) jointly determine the geographical distribution patterns of microorganisms in ice-covered lakes, with stochastic processes dominating. These results expand the understanding of microbial diversity, distribution patterns, and metabolic processes in polar ice-covered lakes.

High-Throughput Screening of Functional Neo-Antigens and Their Specific T-Cell Receptors via the Jurkat Reporter System Combined with Droplet Microfluidics.

T-cell receptor (TCR)-engineered T cells can precisely recognize a broad repertoire of targets derived from both intracellular and surface proteins of tumor cells. TCR-T adoptive cell therapy has shown safety and promising efficacy in solid tumor immunotherapy. However, antigen-specific functional TCR screening is time-consuming and expensive, which limits its application clinically. Here, we developed a novel integrated antigen-TCR screening platform based on droplet microfluidic technology, enabling high-throughput peptide-major histocompatibility complex (pMHC)-to-TCR paired screening with a high sensitivity and low background signal. We introduced DNA barcoding technology to label peptide antigen candidate-loaded antigen-presenting cells and Jurkat reporter cells to check the specificity of pMHC-TCR candidates. Coupled with the next-generation sequencing pipeline, interpretation of the DNA barcodes and the gene expression level of the Jurkat T-cell activation pathway provided a clear peptide-MHC-TCR recognition relationship. Our proof-of-principle study demonstrates that the platform could achieve pMHC-TCR paired high-throughput screening, which is expected to be used in the cross-reactivity and off-target high-throughput paired testing of candidate pMHC-TCRs in clinical applications.

Recent developments in applications of physical fields for microbial decontamination and enhancing nutritional properties of germinated edible seeds and sprouts: a review.

Germinated edible seeds and sprouts have attracted consumers because of their nutritional values and health benefits. To ensure the microbial safety of the seed and sprout, emerging processing methods involving physical fields (PFs), having the characteristics of high efficiency and environmental safety, are increasingly proposed as effective decontamination processing technologies. This review summarizes recent progress on the application of PFs to germinating edible seeds, including their impact on microbial decontamination and nutritional quality and the associated influencing mechanisms in germination. The effectiveness, application scope, and limitation of the various physical techniques, including ultrasound, microwave, radio frequency, infrared heating, irradiation, pulsed light, plasma, and high-pressure processing, are symmetrically reviewed. Good application potential for improving seed germination and sprout growth is also described for promoting the accumulation of bioactive compounds in sprouts, and subsequently enhancing the antioxidant capacity under favorable PFs processing conditions. Moreover, the challenges and future directions of PFs in the application to germinated edible seeds are finally proposed. This review also attempts to provide an in-depth understanding of the effects of PFs on microbial safety and changes in nutritional properties of germinating edible seeds and a theoretical reference for the future development of PFs in processing safe sprouted seeds.

Mixed organic and inorganic amendments enhance soil microbial interactions and environmental stress resistance of Tibetan barley on plateau farmland.

Sufficient crop yield while maintaining soil health and sustainable agricultural development is a global objective, serving a special challenge to certain climate-sensitive plateau areas. Despite conducting trails on a variety of soil amendments in plateau areas, systematic research is lacking regarding the influences of organic and inorganic amendments on soil quality, particularly soil microbiome. To our knowledge, this was the first study that compared the effects of inorganic, organic, and mixed amendments on typical plateau crop hulless barley (Hordeum vulgare L. var. Nudum, also known as "Qingke" in Chinese) over the course of tillering, jointing, and ripening. Microbial communities and their responses to amendments, soil properties and Tibetan hulless barley growth, yield were investigated. Results indicated that mixed organic and inorganic amendments promoted the abundance of rhizosphere microorganisms, enhancing the rhizosphere root-microbes interactions and resistance to pathogenic bacteria and environmental stresses. The rhizosphere abundant and significantly different genera Arthrobacter, Rhodanobacter, Sphingomona, Nocardioides and so on demonstrated their unique adaptation to the plateau environment based on the results of metagenomic binning. The abundance of 23 genes about plant growth and environmental adaptations in the mixed amendment soil were significantly higher than other treatments. Findings from this study suggest that the mixed organic/inorganic amendments can help establish a healthy microbiome and increase soil quality while achieving sufficient hulless barley yields in Tibet and presumably other similar geographic areas of high altitude.

CuO Nanoparticles Alter the Rhizospheric Bacterial Community and Local Nitrogen Cycling for Wheat Grown in a Calcareous Soil.

The application of nanoparticles (NPs) to soils, as either fertilizers or fungicides (e.g., CuO NPs), has been proposed to improve the sustainability of agriculture. The observed effects could result directly from the NP-plant interactions or indirectly through effects on the soil microbiome. The objective of this study was to assess the effects of CuO NPs on the changes in the bacterial community structure and nitrogen-cycling-associated functions in a high pH soil and to correlate these changes with nitrate accumulation, soil parameter changes, and plant growth over 28 days. Triticum aestivum seedlings were exposed to 50 mg/kg CuO NPs, 50 mg/kg CuSO4, or 0.5 mg/kg CuSO4 in a standard soil (Lufa 2.1 soil, pH adjusted to 7.6). While Cu treatments reduced nitrate accumulation in the bulk soil, the effects were opposite in the rhizosphere (the soil influenced by root exudates). While nitrate accumulation in bulk soil negatively correlated with total Cu concentration, part of the nitrate concentration in the rhizosphere was explained by root uptake during plant growth, the rest being modulated by Cu treatments. The abundance of genes involved in the nitrogen cycle in the rhizosphere soil correlated with the ionic copper concentration. The increased nitrate concentration in the rhizosphere correlated with an increase of the gene abundance related to the nitrogen fixation and a decrease of denitrification gene abundance. Microbial diversity in bulk or rhizosphere soil under the different treatments alone could not explain these variations, while differences in the assemblages of bacteria associated with these functional gene abundances gave good insights. This study highlights the complexity of microbial N-related function in the rhizosphere and the need to characterize the rhizosphere soil, plant growth and root activity, NP (bio)transformations, along with microbial networks, to understand the impact of agrochemicals (here CuO NPs) on soil fertility.

Mechanism of 1,4-dioxane microbial degradation revealed by 16S rRNA and metatranscriptomic analyses.

1,4-Dioxane (dioxane), a probable human carcinogen, often exists in industrial wastewater and domestic sewage. In this study, we applied 16S rRNA and metatranscriptomic methods to analyze the dioxane biodegradation mechanism by activated sludge. Tetrahydrofuran (THF) was added as an essential co-metabolite to promote the degradation of dioxane. We found the dioxane removal ratio increased with increasing THF concentrations. When the THF concentration increased from 60.0 to 200.0 mg/L, the dioxane degradation rate was stable. Three additions of ∼60.0 mg/L THF resulted in better dioxane degradation than one addition of 200 mg/L THF. Ammonia-oxidizing and denitrifying bacteria with methane monooxygenases (MOs) and ammonia MOs played the most important roles during the degradation of dioxane. Kyoto Encyclopedia of Genes and Genomes metabolic pathway and functional genes analyses showed that the activated sludge system was complex and stable when dioxane was added. In future studies, primers should be designed to identify specific bacteria and functional MO genes, which would help reveal the function of various bacteria and their MOs during dioxane degradation.

Diversity and arsenic-tolerance potential of bacterial communities from soil and sediments along a gold tailing contamination gradient.

Gold tailings often release arsenic (As) contaminants into the surrounding environment. Microorganisms play an important role in the As cycle, whereas the effects of As on bacterial communities remain unclear. To reveal the effects of As on the diversity of bacterial communities and their As-tolerance potential, farmland soil and river sediment samples were collected at various distances from tailings in the Dandong area of northeastern China. The bacterial communities were analyzed using high-throughput sequencing of 16S rRNA genes. The membrane transport proteins ArsB and (or) ACR3 pump As(III) out of the cell to resist As toxicity. We studied the abundance and phylogeny of ArsB and ACR3 using PCR-based clone libraries and quantitative PCR. The bacterial community was divided into 10 phyla and 59 genera. The transformation from As(V) to As(III) was predominant, which was coupled with denitrification. Both ArsB and ACR3 likely evolved from different orders of Proteobacteria. The arsB gene seems to be more stable in bestowing bacteria with the capability to respond to the As concentration. Moreover, As with iron, manganese, and total organic carbon also influenced the clustering relationships of samples and bacterial distribution.

Improved ethyl caproate production of Chinese liquor yeast by overexpressing fatty acid synthesis genes with OPI1 deletion.

During yeast fermentation, ethyl esters play a key role in the development of the flavor profiles of Chinese liquor. Ethyl caproate, an ethyl ester eliciting apple-like flavor, is the characteristic flavor of strong aromatic liquor, which is the best selling liquor in China. In the traditional fermentation process, ethyl caproate is mainly produced at the later fermentation stage by aroma-producing yeast, bacteria, and mold in a mud pit instead of Saccharomyces cerevisiae at the expense of grains and fermentation time. To improve the production of ethyl caproate by Chinese liquor yeast (S. cerevisiae) with less food consumption and shorter fermentation time, we constructed three recombinant strains, namely, α5-ACC1ΔOPI1, α5-FAS1ΔOPI1, and α5-FAS2ΔOPI1 by overexpressing acetyl-CoA carboxylase (ACC1), fatty acid synthase 1 (FAS1), and fatty acid synthase 2 (FAS2) with OPI1 (an inositol/choline-mediated negative regulatory gene) deletion, respectively. In the liquid fermentation of corn hydrolysate, the contents of ethyl caproate produced by α5-ACC1ΔOPI1, α5-FAS1ΔOPI1, and α5-FAS2ΔOPI1 increased by 0.40-, 1.75-, and 0.31-fold, correspondingly, compared with the initial strain α5. The contents of other fatty acid ethyl esters (FAEEs) (C8:0, C10:0, C12:0) also increased. In comparison, the content of FAEEs produced by α5-FAS1ΔOPI1 significantly improved. Meanwhile, the contents of acetyl-CoA and ethyl acetate were enhanced by α5-FAS1ΔOPI1. Overall, this study offers a promising platform for the development of pure yeast culture fermentation of Chinese strong aromatic liquor without the use of a mud pit.

Inverted tetrahedron-pyramidal micropatterned polymer films for boosting light output power in flip-chip light-emitting diodes.

We report the improved light output power in gallium nitride-based green flip-chip light-emitting diodes (FCLEDs) employed with inverted tetrahedron-pyramidal micropatterned polydimethylsiloxane (ITPM PDMS) films as an encapsulation and protection layer. The micropatterns are transferred into the surface of PDMS films from the sapphire substrate master molds with two-dimensional periodic hexagonal TPM arrays by a soft imprint lithography method. The ITPM PDMS film laminated on the sapphire dramatically enhances the diffuse transmittance (T(D)) in a wavelength (λ) range of 400-650 nm, exhibiting the larger T(D) value of ~53% at λ = 525 nm, (cf., T(D) ~1% for planar sapphire). By introducing the ITPM PDMS film on the outer surface of sapphire in FCLEDs, the light output power is enhanced, indicating the increment percentage of ~11.1% at 500 mA of injection current compared to the reference FCLED without the ITPM PDMS film, together with better electroluminescence intensity and far-field radiation pattern.

Broadband high-reflective distributed Bragg reflectors based on amorphous silicon films for semiconductor laser facet coatings.

We fabricated amorphous silicon (a-Si)-based distributed Bragg reflectors (DBRs) consisting of alternating dense/porous films (i.e., pair) for a center wavelength (λ(c)) of 0.96 µm by oblique angle deposition (OAD) technique using an electron-beam evaporation system. The dense (high refractive index, i.e., high-n) and porous (low-n) a-Si films were deposited at two incident vapor flux angles of 0° and 80° in the OAD, respectively. Their optical reflectance characteristics were investigated in the wavelength range of 0.6-1.5 µm, including theoretical comparison using a rigorous coupled-wave analysis method. Above three pairs, the reflectivity (R) of a-Si DBRs was almost saturated at wavelengths around 0.96 µm, exhibiting R values of >97%. For the a-Si DBR with only three pairs, a broad normalized stop bandwidth (Δλ/λ(c)) of ∼22.5% was obtained at wavelengths of ∼0.87-1.085 µm, keeping high R values of >95%. To simply demonstrate the feasibility of device applications, the a-Si DBR with three pairs was coated as a high-reflection layer at the rear facet of GaAs/InGaAs quantum-well laser diodes (LDs) operating at λ=0.96 µm. For the LDs coated with three-pair a-Si DBR, external differential quantum efficiency (η(d)) was nearly doubled compared to the uncoated LDs, indicating the η(d) value of ∼50.6% (i.e., η(d)∼25.5% for the uncoated LDs).

Tunable distributed Bragg reflectors with wide-angle and broadband high-reflectivity using nanoporous/dense titanium dioxide film stacks for visible wavelength applications.

Highly-tolerant distributed Bragg reflectors (DBRs) based on the same materials consisting of nanoporous/dense titanium dioxide (TiO2) film pair structures with wide-angle and broadband highly-reflective properties at visible wavelengths are reported. For a high refractive index contrast, the two dense and nanoporous TiO2 film stacks are alternatingly deposited on silicon (Si) substrates by a oblique angle deposition (OAD) method at two vapor flux angles (θα) of 0 and 80° for high and low refractive indices, respectively. For the TiO2 DBRs at a center wavelength (λ(c)) of 540 nm, the maximum level in reflectance (R) band is increased with increasing the number of pairs, exhibiting high R values of > 90% for 5 pairs, and the normalized stop bandwidth (∆λ/λ(c)) of ~17.8% is obtained. At λ(c) = 540 nm, the patterned TiO2 DBR with 5 pairs shows an uniform relative reflectivity over a whole surface of 3 inch-sized Si wafer and a large-scalable fabrication capability with any features. The angle-dependent reflectance characteristics of TiO2 DBR at λ(c) = 540 nm are also studied at incident angles (θ(inc)) of 20-70° for p-, s-, and non-polarized lights in the wavelength region of 350-750 nm, yielding high R values of > 70.4% at θ(inc) values of 20-70° for non-polarized light. By adjusting the λ(c)/4 thicknesses of nanoporous and dense films, for λ(c) = 450, 540, and 680 nm, tunable broadband TiO2 DBRs with high R values of > 90% at wavelengths of 400-800 nm are realized.

Composition and variation of sediment bacterial and nirS-harboring bacterial communities at representative sites of the Bohai Gulf coastal zone, China.

With rapid urbanization, anthropogenic activities are increasingly influencing the natural environment of the Bohai Bay. In this study, the composition and variation of bacterial and nirS-harboring bacterial communities in the coastal zone sediments of the Bohai Gulf were analyzed using PCR-based clone libraries. A total of 95 genera were detected in the bacterial communities, with Proteobacteria (72.1 %), Acidobacteria (10.5 %), Firmicutes (1.7 %), Bacteroidetes (1.4 %), Chloroflexi (0.7 %) and Planctomycetes (0.7 %) being the dominated phyla. The NirS sequences were divided into nine Clusters (A-I). Canonical correlation analysis showed that the bacterial or denitrifying communities were correlated with different environmental factors, such as total organic carbon, total nitrogen, ammonium, sulfate, etc. Furthermore, bacterial communities' composition and diversity are influenced by oil exploration, sewage discharge and other anthropogenic activities in the coastal area of the Bohai Sea. Thus, this study provided useful information on further research on regional or global environmental control and restore.

Critical role of multiple antibiotics on the denitrification rate in groundwater: Field investigative proof.

The impact of antibiotics on denitrification has emerged as a significant topic; however, there is a dearth of mechanistic understanding regarding the effects of multiple antibiotics at the ng/L level on denitrification in groundwater. This study conducted five field samplings between March 2019 and July 2021 at two representative monitoring wells. The investigation utilized metagenomic sequencing to unveil the antibiotic mechanisms influencing denitrification. Results revealed the detection of 16 out of 64 antibiotics, with a maximum detection frequency and total concentration of 100 % and 187 ng/L, respectively. Additionally, both nitrate and total antibiotic concentrations exhibited a gradual decrease along the groundwater flow direction. Metagenomic evidence indicated that denitrification served as the dominant biogeochemical process controlling nitrate attenuation in groundwater. However, the denitrification capacity experienced significant inhibition in the presence of multiple antibiotics at the ng/L level. This inhibition was attributed to decreases in the relative abundance of dominant denitrifying bacteria (Candidatus_Scalindua, Herminiimonas and unclassified_p_Planctomycetes) and denitrifying functional genes (narGH, nirKS and norB), signifying the pressure exerted by antibiotics on denitrifying bacteria. The variation in antibiotic concentration (∆Cantibiotics) indicated a change in antibiotic pressure on denitrifying bacteria. A larger ∆Cantibiotics corresponded to a greater rebound in the relative abundance of denitrifying functional genes, resulting in a faster denitrification rate (Kdenitrification). Field observations further demonstrated a positive correlation between Kdenitrification and ∆Cantibiotics. Comparatively, a higher Kdenitrification observed at higher ∆Cantibiotics was primarily due to the enrichment of more nondominant denitrifying bacteria carrying key denitrifying functional genes. In conclusion, this study underscores that multiple antibiotics at the ng/L level in groundwater inhibited denitrification, and the degree of inhibition was closely related to ∆Cantibiotics.

Revealing discrepancies and drivers in the impact of lomefloxacin on groundwater denitrification throughout microbial community growth and succession.

The coexistence of antibiotics and nitrates has raised great concern about antibiotic's impact on denitrification. However, conflicting results in these studies are very puzzling, possibly due to differences in microbial succession stages. This study investigated the effects of the high-priority urgent antibiotic, lomefloxacin (LOM), on groundwater denitrification throughout microbial growth and succession. The results demonstrated that LOM's impact on denitrification varied significantly across three successional stages, with the most pronounced effects exhibited in the initial stage (53.8% promotion at 100 ng/L-LOM, 84.6% inhibition at 100 µg/L-LOM), followed by the decline stage (13.3-18.2% inhibition), while no effect in the stable stage. Hence, a distinct pattern encompassing susceptibility, insusceptibility, and sub-susceptibility in LOM's impact on denitrification was discovered. Microbial metabolism and environment variation drove the pattern, with bacterial numbers and antibiotic resistance as primary influencers (22.5% and 15.3%, p < 0.01), followed by carbon metabolism and microbial community (5.0% and 3.68%, p < 0.01). The structural equation model confirmed results reliability. Bacterial numbers and resistance influenced susceptibility by regulating compensation and bacteriostasis, while carbon metabolism and microbial community impacted energy, electron transfer, and gene composition. These findings provide valuable insights into the complex interplay between antibiotics and denitrification patterns in groundwater.

The antibodies against the A137R protein drive antibody-dependent enhancement of African swine fever virus infection in porcine alveolar macrophages.

African swine fever virus (ASFV) is the causative agent of African swine fever (ASF), a highly contagious disease that can kill up to 100% of domestic pigs and wild boars. It has been shown that the pigs inoculated with some ASF vaccine candidates display more severe clinical signs and die earlier than do pigs not immunized. We hypothesize that antibody-dependent enhancement (ADE) of ASFV infection may be caused by the presence of some unidentified antibodies. In this study, we found that the ASFV-encoded structural protein A137R (pA137R) can be recognized by the anti-ASFV positive sera, indicating that the anti-pA137R antibodies are induced in the ASFV-infected pigs. Interestingly, our results demonstrated that the anti-pA137R antibodies produced in rabbits or pigs enhanced viral replication of different ASFV strains in primary porcine alveolar macrophages (PAMs), the target cells of ASFV. Mechanistic investigations revealed that anti-pA137R antibodies were able to promote the attachment of ASFV to PAMs and two types of Fc gamma receptors (FcγRs), FcγRII and FcγRIII, mediated the ADE of ASFV infection. Taken together, anti-pA137R antibodies are able to drive ASFV ADE in PAMs. These findings shed new light on the roles of anti-ASFV antibodies and have implications for the pathophysiology of the disease and the development of ASF vaccines.

Soil bacterial communities shaped by geochemical factors and land use in a less-explored area, Tibetan Plateau.

BACKGROUND: As the largest low-latitude permafrost region, the Tibetan Plateau (TP) is an important part of the earth's terrestrial ecosystem and one of the most vulnerable areas to climate change and human activities. However, to the best of our knowledge, the bacterial communities in TP soils and their roles in biogeochemical cycles remain limited. RESULTS: In this study, we report the bacterial community structure and function as well as their correlation with environmental factors in TP major ecosystems (farmland, alpine meadow and oligosaline lake) by using metagenomic approaches. Compared with other soil samples in various environments, TP soils share a core set of microorganisms with a distinct abundance and composition. Among TP soil samples, the taxonomic and functional composition of bacterial communities among the upper (3-5 cm) and lower (18-20 cm) soils of farmland sites were highly similar, whereas the dissimilarities within alpine meadow samples were significantly greater than among farmland samples. A similar pattern was observed in elements cycles and pathways associated with adaption to environment and land use types. Canonical correlation analysis revealed that the bacterial communities in most of farmland and alpine meadow soil samples were also significantly correlated with geogenic variables. Specifically, the root-nodule bacteria are negatively correlated with the soil moisture and pH, while Thiobacillus associated with sulfur cycles show potential responses to low temperature and intense UV radiation. CONCLUSIONS: These findings indicate that the bacterial community structure and functions in TP soils were influenced by both human activities and soil environmental properties, and that the bacterial communities appeared to be more homogenized in the farmland soils compared with pristine alpine meadows.

Determination of trace chromium (VI) using a hollow-core metal-cladding optical waveguide sensor.

A biosensor capable of highly sensitive detection of trace chromium (VI) with a simple hollow-core metal-cladding waveguide (HCMW) structure is theoretically modeled and experimentally demonstrated. Owing to the high sensitivity of the excited ultrahigh-order modes in the waveguide, a tiny variation of the extinction coefficients in the waveguide guiding layer where the chromate ions reacts with the diphenylcarbazide (DPC) can lead to a significant change of light intensity in the reflection spectrum. The experimental results indicate that using the proposed method, the chromium (VI) sensitivity detection limit can be as low as 1.2 nM, which represents a 16-fold improvement compared to the surface plasmon field-enhanced resonance light scattering (SP-RLS) method, and a 4-fold improvement compared to the flame atomic absorption spectrometry and fluorimetry spectroscopy, respectively.

Optimization of cold-active lipase production from psychrophilic bacterium Moritella sp. 2-5-10-1 by statistical experimental methods.

Statistical experimental designs were applied to optimize cold-active lipase production by the psychrophilic bacterium Moritella sp. 2-5-10-1. First, a Plackett-Burmen design (PBD) was used to evaluate the significant effects of various fermentation parameters. The results indicated that soybean meal, temperature, and Tween-80 had significant influences on lipase production. The levels of these variables were optimized subsequently using central composite design (CCD). A quadratic regression model of cold-active lipase production was built, and verification experiments confirmed its validity. On subsequent scale-up in a 10-L bioreactor using optimized conditions, cold-active lipase production (30.56 U/mL) was obtained. The results clearly indicated that the model was adequate even on a large scale. To our knowledge, this is the first report of statistical optimization of cold-active lipase production by a psychrophilic bacterium.

Microbiota associated with the migration and transformation of chlorinated aliphatic hydrocarbons in groundwater.

Pollution of groundwater with chlorinated aliphatic hydrocarbons (CAHs) is a serious environmental problem which is threatening human health. Microorganisms are the major participants in degrading these contaminants. Here, groundwater contaminated for a decade with CAHs was investigated. Numerical simulation and field measurements were used to track and forecast the migration and transformation of the pollutants. The diversity, abundance, and possible activity of groundwater microbial communities at CAH-polluted sites were characterized by molecular approaches. The number of microorganisms was between 5.65E+05 and 1.49E+08 16S rRNA gene clone numbers per liter according to quantitative real-time PCR analysis. In 16S rRNA gene clone libraries constructed from samples along the groundwater flow, eight phyla were detected, and Proteobacteria were dominant (72.8 %). The microbial communities varied with the composition and concentration of pollutants. Meanwhile, toluene monooxygenases and methane monooxygenases capable of degradation of PCE and TCE were detected, demonstrating the major mechanism for PCE and TCE degradation and possibility for in situ remediation by addition of oxygen in this study.