Emergence of mcr-4.3 genes in a novel Shewanella specie isolated from the Arctic environment.

Mobile colistin resistance (mcr) genes are pivotal contributors to last-line of antimicrobial resistance in human infections. Shewanella, historically recognized as a natural environmental bacterium with metal reduction capabilities, recently has been observed in clinical settings. However, limited knowledge has been explored on genetic differences between strains from non-clinical and clinical strains. In this study, we conducted the whole genome sequencing on six Arctic strains, illustrated the phylogenetic relationships on published 393 Shewanella strains that categorized the genus into four lineages (L1 to L4). Over 86.4% of clinical strain group (CG) strains belonged to L1 and L4, carrying mcr-4 genes and a complete metal-reduction pathways gene cluster. Remarkably, a novel Arctic Shewanella strain in L3, exhibits similar genetic characteristics with CG strains that carried both mcr-4 genes and a complete metal reduction pathway gene cluster. It raised concerns about the transmission ability from environment to clinic setting causing in the potential infections, and emphasized the need for monitoring the emerging strains with human infections.

Comparison of orbital fibroblasts from Graves' ophthalmopathy and healthy control.

Graves' ophthalmopathy (GO) is an extrathyroidal manifestation of Graves' disease, Orbital fibroblasts (OFs) are recognized as key players in GO pathogenesis, involved in orbital inflammation, tissue remodeling, and fibrosis. This study offers a primary exploration of cell behavior and characteristics on OFs from GO (GO-OFs), and compared to OFs from healthy control (HC-OFs). Results reveal that GO-OFs exhibit delayed migration from tissue fragments, while no significant difference in cell proliferation is observed between GO-OFs and HC-OFs. Aberrant expression pattern of surface proteins Thy-1, TSHR, and IGF-1R suggests shared autoantigens and pathways between GO and GD, contributing to inflammation and fibrosis. Investigations into cytokine responses unveil elevated secretion of hyaluronic acid (HA) and prostaglandin E2 (PGE2) in GO-OFs, emphasizing their role in tissue remodeling. These findings deepen our understanding of OFs in GO pathogenesis, offering potential therapeutic avenues.

Prolonged Button Battery Exposure Leading to Severe Ocular Injury without Heavy Metal Poisoning.

Introduction: Prolonged exposure to a complete button battery can cause severe tissue necrosis in the eye and permanent impairment of visual function. The main mechanism of injury is the current generated by the hydrolysis of tissue fluid at the negative electrode and the production of hydroxide ions. Case Presentation: A 3-year-old girl went to the local hospital because of swelling and pain in her right eye of 12-h duration. The local doctor performed an orbital CT (computed tomography) scan and found a foreign body between the right eyelid and the eyeball. The foreign body was removed immediately under general anesthesia. In addition, it was found that the foreign body was a button battery, but it prolonged 39 h from the onset of the child's symptoms. The child underwent a second operation in our hospital and received amniotic membrane transplantation combined with conjunctival flap coverage. Topical corticosteroid and antibiotic eye ointment were continued for 3 months after surgery. Local pigmentation was seen, there was no symblepharon, but the cornea was still opaque and the visual acuity was only FC (finger count). In this particular case, heavy metal testing conducted on the child's blood fortunately revealed that the levels were within the normal range. Conclusion: Early detection and urgent removal of button battery are crucial in order to minimize exposure time. We should also be concerned about heavy metals in the blood. Children should be kept away from button batteries as much as possible to avoid such injury.

Identification of Pathogens and Evaluation of Resistance and Genetic Diversity of Maize Inbred Lines to Stalk Rot in Heilongjiang Province, China.

Maize stalk rot, caused by multiple pathogens, is a serious soilborne disease worldwide. Composition of pathogens causing maize stalk rot and resistance of maize inbred lines in Heilongjiang Province, China, are not well understood. In this study, 138 fungal isolates were collected from different maize-producing areas in Heilongjiang Province, which were identified as Fusarium graminearum (23.2%), F. subglutinans (18.9%), F. cerealis (18.9%), Bipolaris zeicola (13.0%), F. brachygibbosum (13.0%), F. temperatum (7.2%), and F. proliferatum (5.8%). Among them, F. graminearum (>20%) was the predominant species among the isolates causing maize stalk rot. B. zeicola had not previously been reported causing maize stalk rot in China. Resistance of 67 maize inbred lines to maize stalk rot was assessed, and 24 lines (35.8% of them) were highly resistant or resistant, indicating that approximately 65% of these lines were susceptible to maize stalk rot. Maize inbred lines were analyzed using simple sequence repeat markers and divided into five genetic groups with 12 pairs of primers. Additionally, analysis of molecular variance indicated that 44.2% of the genetic variation in disease resistance was distributed among populations. This study provides insight into the genetic diversity of inbred maize and may contribute useful information for breeding stalk rot disease-resistant hybrids, and facilitates development of effective strategies for managing this destructive disease complex.

Influence of environmental factors on soil organic carbon in different soil layers for Chinese Mollisols under intensive maize cropping.

The Mollisol region of Northeast China has a large soil organic carbon (SOC) storage which is important for maintaining soil fertility. SOC is susceptible to various environmental factors; however, the responses of SOC content to environmental factors in different soil layers of cropland remain unclear, particularly in deep soil layers. In this study, we collected 138 soil samples from the surface, subsurface, and subsoil layers among 46 sample sites with monocropping maize and intensive conventional tillage in this region. We assessed the relative importance and effect paths of 12 environmental factors (including geography, climate, and soil properties) on SOC content in different layers using redundancy analysis (RDA), structural equation model (SEM), and variation partitioning analysis (VPA). The VPA results showed that SOC content was mainly affected by climatic factors that explained 68% and 57% for the surface and subsurface layers, respectively. However, SOC content in the subsoil layer was greatly affected by soil properties that explained 27%. Furthermore, the SEMs results suggested that geographical factors indirectly affected SOC content by influencing the climatic factors. Mean annual temperature was the most important factor affecting SOC content directly or indirectly, and its negative effects significantly diminished with soil depth, as it explained 63%, 52%, and 17% of the variation in SOC content for the surface, subsurface and subsoil layers, respectively. In addition, the effects of soil water-holding capacity on SOC content also decreased with soil depth, whereas pH and clay content showed a contrasting pattern. This implies that pH and clay content play important roles in the sequestration of SOC in deep soil layers. Moreover, the organic C content within >53 µm aggregates was more sensitive to environmental factors. This study can be useful for forecasting SOC dynamics and establishing reasonable C management strategies under climate change conditions.

Response of the chemical structure of soil organic carbon to modes of maize straw return.

Elucidating the chemical structure of soil organic matter (SOM) is important for accurately evaluating the stability and function of SOM. Aboveground vegetation directly affects the quantity and quality of exogenous organic matter input into the soil through plant residues and root exudates, which in turn affects soil microbial species, community structure, and activity, and ultimately impacts the chemical structure of SOM. In this study, a 13C nuclear magnetic resonance technique was used to analyze the chemical structure characteristics of soil organic carbon (SOC) under various rates of straw returning combined with rotary tillage and under full straw mulching. The results showed that full straw returning with rotary tillage and full straw mulching more effectively increased the SOC content than reduced rate of straw returning (1/2 and 1/3 of full straw) with rotary tillage. The contents of alkyl C and alkoxy C in the functional groups of SOC under various straw returning treatments were increased compared with those under the treatment of maize stubble remaining in soil (CK). Furthermore, the contents of aromatic C and carboxyl C were decreased, which were consistent with the chemical shift changes of SOC. Compared with CK treatment, straw returning decreased the content of aromatic C in the functional groups of SOC, but increased the content of alkoxy C, which could be associated with the change in integral areas of absorption peaks of alkyl C and alkoxy C moving toward left and right, respectively. The content of total SOC was significantly positively (P < 0.05) correlated with that of alkoxy C and significantly negatively (P < 0.01) correlated with that of aromatic C. The molecular structure of SOC tends to be simplified due to the decreasing in refractory C and the increasing in easily decomposed C after straw returning to the field.

Investigations of the effect of the amount of biochar on soil porosity and aggregation and crop yields on fertilized black soil in northern China.

The combination of chemical fertilizer and biochar is regarded as a useful soil supplement for improving the properties of soil and crop yields, and this study describes how the biochar of maize straw can be used to improve the quality of the degraded black soil. This has been achieved by examining the effects of combining different amounts of biochar with chemical fertilizer on the porosities and aggregate formation of soil and exploring how these changes positively impact on crop yields. A field trial design combining different amounts of maize straw biochar [0 (NPK), 15.75 (BC1), 31.5 (BC2), and 47.25 t ha-1 (BC3)] with a chemical fertilizer (NPK) has been used to investigate changes in the formation of soil aggregate, clay content, soil organic carbon (SOC), and crop yields in Chinese black soil over a three year period from 2013 to 2015. The results of this study show that the addition of fertilizer and biochar in 2013 to black soil results in an increased soybean and maize yields from 2013 to 2015 for all the treatments, with BC1/BC2 affording improved crop yields in 2015, while BC3 gave a lower soybean yield in 2015. Total porosities and pore volumes were increased for BC1 and BC2 treatments but relatively decreased for BC3, which could be attributed to increased soil capillary caused by the presence of higher numbers of fine soil particles. The addition of biochar had a positive influence on the numbers and mean weight diameters (MWD) of soil macroaggregates (>0.25 mm) that were present, with the ratio of SOC to TN in soil macroaggregates found to be greater than in the microaggregates. The most significant amount of carbon present in macroaggregates (>2 mm and 0.25-2 mm) was observed when BC2 was applied as a soil additive. Increasing the levels of maze straw biochar to 47.25 t ha-1 led to an increase in the total organic carbon of soil, however, the overall amount of macroaggregates and MWD were decreased, which is possibly due to localized changes in microbial habitat. The supplementation of biochar increased in the amount of aromatic C present (most significant effect observed for BC2), with the ratio of aliphatic C to aromatic C found to be enhanced due to a relative reduction in the aliphatic C content with >2 mm particle fraction. These changes in organic carbon content and soil stability were analyzed using univariate quadratic equations to explain the relationship between the type of functional groups (polysaccharide C, aliphatic C, aromatic C, aliphatic C/aromatic C) present in the soil aggregates and their MWDs, which were found to vary significantly. Overall, the results of this study indicate that the use of controlled amounts of maize-straw biochar in black soil is beneficial for improving crop yields and levels of soil aggregation, however, the use of excessive amounts of biochar results in unfavorable aggregate formation which negatively impacts the yields of crop growth. The data produced suggest that aromatic C content can be used as a single independent variable to characterize the stability of soil aggregate when biochar/fertilizer mixtures are used as soil additives to boost growth yields. Analysis of soil and crop performance in black soil revealed that the application of maize-straw biochar at a rate of 15.75 and 31.5 t ha-1 had positive effects on crop yields, soil aggregation and accumulation of aromatic C in the aggregate fractions when a soybean-maize rotation system was followed over three years.

Impacts of Long-term Fertilization on the Molecular Structure of Humic Acid and Organic Carbon Content in Soil Aggregates in Black Soil.

Soil aggregates are the basic units of soil structure, and their composition and carbon (C) stability directly affect soil fertility. As cementing agents, humic substances play an important role in the formation and stability of soil aggregates. Long-term fertilization not only changes the structure of humic acid (HA), but also affects the content and stability of organic C in soil aggregates. In this study, based on a long-term fertilization experiment, the relationship between the molecular structure of HA and the stability of organic C in the aggregates was examined. Compared with the non-fertilization control (CK), both the application of organic manure alone (M) and organic manure combined with inorganic fertilizer application (MNPK) increased organic C content in the bulk soil and in HA. In addition, the application of organic manure (M, MNPK) favored the formation of macroaggregates (>0.25 mm) and showed a higher organic C contents of soil aggregates with different sizes than the CK. Moreover, the content of aliphatic C, the ratios of aliphatic C/aromatic C and alkyl C/O-alkyl C was increased with the application of organic fertilizer. A significant negative correlation was observed between aromatic C and organic C content of the aggregates with sizes of >2 mm, 2-0.25 mm, and 0.25-0.053 mm (P < 0.05). The findings indicated that organic fertilization treatments (M and MNPK) increased the aliphatic C content of HA, which favored the increase in the organic C content and stability of the aggregates.

Responses of Labile Organic Nitrogen Fractions and Enzyme Activities in eroded Mollisols After 8-year Manure Amendment.

Soil erosion will cause a degradation in soil nitrogen supplying capacity (SNSC) and manure amendment is an effective way to restored eroded soils. Both labile fractions of soil organic N (SON) and N transformation enzymes are indicators for SNSC, but the effect of manure amendments on labile SON fractions and the relationship between labile SON fractions and enzyme activities remains unclear. In this study, five degrees of erosion were simulated in Mollisols (removal of 0, 5, 10, 20 and 30 cm of topsoil) to analyse the changes in labile SON fractions and nitrogen transformation enzyme activities after 8-year manure amendment. We found that soil total N (TN), labile SON fractions and enzyme activities all increased after manure amendments. The largest labile SON fraction was particle organic nitrogen (POM-N) and the second was light fraction organic nitrogen (LFOM-N), which accounted >60% for TN in total. Correlation analysis showed that both urease and protease activities were significantly correlated with POM-N, LFOM-N, microbial biomass N and dissolvable organic N, indicating that both urease and protease activities can be used to predict labile SON pools and enzyme activities worked similarly in indicating SNSC with labile SON fractions. Altogether, 8-year manure amendment could recover SNSC of lightly eroded Mollisols to natural levels, i.e. erosion depths at 5 cm and 10 cm; however, it is not able to recover SNSC in Mollisols suffering severe erosion.

Effects of long-term fertilization on soil humic acid composition and structure in Black Soil.

The composition and structure of humic acid (HA) can be affected by fertilization, but the short-term effects are difficult to detect using traditional analysis methods. Using a 35-year long-term experiment in Black Soil, the molecular structure of HA was analyzed with Fourier transform infrared spectroscopy (FTIR), 13C nuclear magnetic resonance spectroscopy (NMR), and fluorescence spectroscopy. Variation in HA was analyzed after long-term fertilization, including fertilization with manure (M), inorganic N, P and K fertilizer (NPK), manure combined with inorganic N, P, and K fertilizer (MNPK), and a no-fertilizer control (CK). The application of each fertilizer treatment increased crop yields compared with the CK treatment, and the MNPK treatment increased crop yield the most. The ratio of main IR absorption peak of HA at 2,920 cm-1 compared with the peak at 2,850 cm-1 (2920/2850) was higher in the NPK and MNPK treatments compared with the CK treatment. The application of manure (MNPK and M treatments) increased the ratio of hydrogen to carbon (H/C) in HA, and raised the ratio of the main IR absorption peak of HA at 2920 cm-1 to that at 1720 cm-1 (2920/1720). Manure treatments also raised the ratio of aliphatic carbon (C) to aromatic C, alkyl C to alkoxy C and hydrophobic C to hydrophilic C and the fluorescence index (f 450/500), but decreased the degree of aromatization of HA, when compared with the CK treatment. The ratio between each type of C in HA was similar among all the fertilizer treatments, but NPK had a lower ratio of H/C and a lower content of aliphatic C compared with the CK treatment. These results indicated that the molecular structure of HA in Black Soil tends to be aliphatic, simpler, and younger after the application of manure. While the application of inorganic fertilizers increased in the degree of condensation of HA and made HA structure complicated. The application of manure alone or combined with inorganic fertilizers may be an effective way to increase crop yield and improve the structure of soil organic matter.

The effect of heavy metal contamination on the bacterial community structure at Jiaozhou Bay, China

Abstract In this study, determination of heavy metal parameters and microbiological characterization of marine sediments obtained from two heavily polluted sites and one low-grade contaminated reference station at Jiaozhou Bay in China were carried out. The microbial communities found in the sampled marine sediments were studied using PCR-DGGE (denaturing gradient gel electrophoresis) fingerprinting profiles in combination with multivariate analysis. Clustering analysis of DGGE and matrix of heavy metals displayed similar occurrence patterns. On this basis, 17 samples were classified into two clusters depending on the presence or absence of the high level contamination. Moreover, the cluster of highly contaminated samples was further classified into two sub-groups based on the stations of their origin. These results showed that the composition of the bacterial community is strongly influenced by heavy metal variables present in the sediments found in the Jiaozhou Bay. This study also suggested that metagenomic techniques such as PCR-DGGE fingerprinting in combination with multivariate analysis is an efficient method to examine the effect of metal contamination on the bacterial community structure.

The effect of heavy metal contamination on the bacterial community structure at Jiaozhou Bay, China.

In this study, determination of heavy metal parameters and microbiological characterization of marine sediments obtained from two heavily polluted sites and one low-grade contaminated reference station at Jiaozhou Bay in China were carried out. The microbial communities found in the sampled marine sediments were studied using PCR-DGGE (denaturing gradient gel electrophoresis) fingerprinting profiles in combination with multivariate analysis. Clustering analysis of DGGE and matrix of heavy metals displayed similar occurrence patterns. On this basis, 17 samples were classified into two clusters depending on the presence or absence of the high level contamination. Moreover, the cluster of highly contaminated samples was further classified into two sub-groups based on the stations of their origin. These results showed that the composition of the bacterial community is strongly influenced by heavy metal variables present in the sediments found in the Jiaozhou Bay. This study also suggested that metagenomic techniques such as PCR-DGGE fingerprinting in combination with multivariate analysis is an efficient method to examine the effect of metal contamination on the bacterial community structure.

Expression of recombinant human α-lactalbumin in milk of transgenic cloned pigs is sufficient to enhance intestinal growth and weight gain of suckling piglets.

Human α-lactalbumin (HLA) has very high nutritional value and important physiological functions during the neonatal period. The peptides derived from HLA provide diverse health benefits including antimicrobial, antiviral, immune-modulating, and antihypertensive effects. Thus, it is worth investigating the effects on offspring development of increasing HLA in milk. In this study, we found that recombinant human α-lactalbumin (rHLA) exhibits efficient inhibition of dipeptidyl peptidase-IV (DPP-IV) activity in an in vitro simulated gastrointestinal digestion system. Using a BAC clone containing the complete HLA gene as a candidate vector, we generated two lines of transgenic cloned sows via somatic cell nuclear transfer that over-expressed rHLA. The average concentrations of rHLA in milk from the two lines of transgenic cloned sows were 2.24 ± 0.71 mg/ml and 2.67 ± 1.29 mg/ml. The feeding experiments revealed that rHLA represses dipeptidyl peptidase-IV (DPP-IV) activity in vivo. Furthermore, the piglets reared by rHLA transgenic cloned sows exhibit better performance in gain of body weight and intestine growth than the control piglets reared by non-transgenic sows. Therefore, these findings indicate that rHLA could serve as a natural precursor for a DPP-IV inhibitor, and the transgenic technology that produced the over-expression of rHLA could be a useful method for pig breeders to improve lactation performance.

Seasonal and spatial variation of Deuteromycetes population in polluted cost of Kiaochow Bay.

Objective: To reveal the relationship between Deuteromycetes community and the environmental in Kiaochow Bay of the Yellow Sea. Methods: Using recorded pollution survey, we used molecular methods to study seasonal and spatial variation of Deuteromycetes community diversity in different polluted waters of Kiaochow Bay of the Yellow Sea, China. Results: Denaturing gradient gel electrophoresis fingerprints varied obviously among different sites of similar level of pollution. Moreover, sequence analysis of recovered dominant bands exhibited the existence of plenty of uncultivable fungi, among which Penicillium was the dominant genus. Furthermore, in heavily polluted estuary, there were abundant animal pathogens such as amoeba and Pythium as well as Deuteromycetes. These discoveries demonstrate that the Deuteromycetes community structure is closely related to marine environment, and are indicative of different level of marine contamination. Conclusion: The relationship between Deuteromycetes community and different level of pollution and seasons varied were closely related.

[Effect of long-term fertilization on microbial community functional diversity in black soil].

In order to study the effects of long-term different fertilization on microbial community functional diversity in arable black. soil, we examined microbial metabolic activities in two soil la- yers (0-20 cm, 20-40 cm) under four treatments (CK, NPK, M, MNPK) from a 35-year continuous fertilization field at the Ministry of Agriculture Key Field Observation Station of Harbin Black Soil Ecology Environment using Biolog-ECO method. The results showed that: in the 0-20 cm soil layer, combined application of organic and inorganic fertilizer(MNPK) increased the rate of soil microbial carbon source utilization and community metabolism richness, diversity and dominance; In the 20-40 cm layer, these indices of the MNPK treatment was lower than that of the NPK treat- ment; while NPK treatment decreased soil microbial community metabolism evenness in both layers. Six groups of carbon sources used by soil microbes of all the treatments were different between the two soil layers, and the difference was significant among all treatments in each soil layer (P < 0.05) , while the variations among treatments were different in the two soil layers. Canonical correspondence analysis (CCA) showed that soil microbial community metabolic function of all the treatments was different between the two soil layers, and there was difference among all treatments in each soil layer, while the influences of soil nutrients on soil microbial community metabolic function of all treatments were similar in each soil layer. It was concluded that long-term different fertilization affected soil microbial community functional diversity in both tillage soil layer and down soil layers, and chemical fertilization alone had a larger influence on the microbial community functional diversity in the 20-40 cm layer.

Molecular detection of Penicillium griseofulvum as the coastal pollution indicator.

Molecular methods were carried out to detect Penicillium griseofulvum, a dominant species related to heavy metal pollution, which was screened from marine contaminated sediments. Based on differences in internal transcribed spacer (ITS) sequences of Penicillium genus and specific isoamyl alcohol oxidase (IAO) sequences, species-specific primers AS1/RS4 and IAO1/IAO2 of Penicillium griseofulvum were designed and synthesized which were then employed in optimized PCR systems. The detection sensitivities were compared through ordinary PCR and nested-PCR using two pairs of primers, respectively. Both primer pairs could exclusively amplify destined DNA fragment from contaminated environmental samples in our researches. As for primers AS1/RS4, the detection sensitivity for spores (pure spore DNA) could be 10 fg/µl and 10 spores, respectively, and the detection sensitivity for the sediments was 10² spores/0.25 g sediments. While the detection sensitivity of IAO1/IAO2 primers was lower than that of AS1/RS4. Despite the difference in detection sensitivity, it is feasible that the species-specific primers could be used as probes for the detection of environmental pollution dominant species, Penicillium griseofulvum, since the frequency of occurrence and amount of this strain could preferably indicate the pollution degree.

[Molecular detection of Penicillium griseofulvum as the coastal pollution indicator].

OBJECTIVE: PCR method was used to detect Penicillium griseofulvum, a dominant species in marine contaminated sediments and thereby to deduce the contamination degree. METHODS: According to differences in internal transcribed space (ITS) sequences of Penicillium genus and specific IAO sequence, we designed species-specific primers AS1/RS4 and IAO1/IAO2 of Penicillium griseofulvum and established the corresponding PCR systems. By using PCR and nested-PCR, the detection sensitivity was compared. RESULTS: The primers could exclusively amplify destined DNA fragment from environment. Using AS1/RS4 as primers, the detection sensitivity could be 10 fg/microL and 10 spores. The detection sensitivity for the sediments was 10(2) spores/0.25 g sediments. While the detection was unsensitive when using IAO1/IAO2 as primers. CONCLUSION: It is feasible that the species-specific primers be used as probes for the detection of environmental pollution dominant species, Penicillium griseofulvum, because the frequency of occurrence and amount of this strain could preferably indicate the pollution degree.

[Study on increasing anti-fungi activity of marine microbes by mixed culture].

The effect of mixed culture on the anti-fungi activity of marine microbes metabolites was evaluated. Based on optimization of monoculture, orthogonal design was used to approach the optimal region of the medium composition of mixed fermentation. The mixed culture showing anti-fungi activity of marine microbes was enriched obviously, compared with monoculture and fermentation mixture. The MIC (minimal inhibitory concentration) reduced from 156 microL/mL, 125 microL/mL by monoculture and 250 microL/mL, 218 microL/mL by mixture of monoculture fermentation to 32 microL/mL, 28 microL/mL, indicate 200% to 300% increase separately in fungi inhibition with mixed culture.