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Background/Aim. Staphylococcus aureus (S. aureus) is a gram-positive pathogen that causes various human diseases. S. aureus causes pneumonia, which is characterized by localized tissue necrosis. The aim of the study was to explore the expression of mouse ?-defensin 3 (mBD-3) induced by S. aureus in mouse lungs and the effect of mBD-3 expression on the mitogen-activated protein kinase (MAPK) pathway. Methods. An experimental model of S. aureus pneumonia in mice was developed, and the expression of mBD-3 and activation of the MAPK pathway were investigated using the methods of immunofluorescence and western blot. Results. The experimental model was created successfully. The number of white blood cells was elevated 48 and 72 hrs after the introduction of bacteria through mouse airways, and bronchiolar mucosal hyperemia was observed, along with a large number of white blood cells and mucus in the bronchioles. The mBD-3 expression levels 48 and 72 hrs after the induction of infection were greater than the levels in the control group and 24 hrs after the induction. The amount of phosphorylated extracellular signal-regulated kinase (ERK1/2) was increased 48 and 72 hrs after infection induction, compared with the levels in the control group and 24 hrs after induction. The expression of mBD-3 was lower when ERK1/2 phosphorylation was inhibited by the U0126 inhibitor. Conclusion. S. aureus in experimental pneumonia mouse model accelerates mBD-3 expression in the mouse lung mainly through an ERK1/2-dependent signaling pathway.

Cyclization of proteins using SpyTag/SpyCatcher is a novel approach to increase their thermal stability. In this paper, we test this approach on two β-galactosidases from Bacillus circulans, BgaB and BgaC, and find that BgaB was stabilized while BgaC was not. Wild-type BgaB precipitated completely upon heating above 70 °C, but after SpyRing cyclization, it remained soluble after heating to 90 °C. Similarly, wild-type BgaB retained only 50 % activity after heating at 60 °C for 10 min, but this increased to 80 % after SpyRing cyclization. In contrast, cyclization decreased the stability of BgaC. After SpyRing cyclization, BgaC only retained 2 % activity after 20-min incubation at 55 °C, whereas the wild-type BgaC retained 25 % activity. One reason for the different effect of cyclization may the shorter distance between the N- and C-termini in BgaB (20.2 Å) as compared to BgaC (43.7 Å). The intrinsic fluorescence and circular dichroism spectra suggested that SpyRing cyclization of BgaB did not significantly change its conformation or secondary structure. SpyRing cyclized BgaB yielded similar amounts and compositions of galacto-oligosaccharides using a high initial lactose concentration (40 %, w/v), but a slightly higher amount at low initial lactose concentration (5 %, w/v) suggesting increased transgalactosylation activity.

Dyed wood is prone to photoaging when exposed to UV irradiation which decreases its decorative effect and service life. Holocellulose, as the main component of dyed wood, has a photodegradation behavior which is still unclear. To investigate the effect of UV irradiation on chemical structure and microscopic morphology changes of dyed wood holocellulose, Maple birch (Betulacostata Trautv) dyed wood and holocellulose were exposed to UV accelerated aging treatment; the photoresponsivity includes crystallization, chemical structure, thermal stability, and microstructure were studied. Results showed that UV radiation has no significant effect on the lattice structure of dyed wood fibers. The wood crystal zone diffraction 2θ and layer spacing was basically unchanged. With the UV radiation time extension, the relative crystallinity of dyed wood and holocellulose showed a trend of increasing first and then decreasing, but the overall change was not significant. The relative crystallinity change range of the dyed wood was not more than 3%, and the dyed holocellulose was not more than 5%. UV radiation caused the molecular chain chemical bond in the non-crystalline region of dyed holocellulose to break, the fiber underwent photooxidation degradation, and the surface photoetching feature was prominent. Wood fiber morphology was damaged and destroyed, finally leading to the degradation and corrosion of the dyed wood. Studying the photodegradation of holocellulose is helpful to understand the photochromic mechanism of dyed wood, and, further, to improve its weather resistance.

Human gut microbiota interfere with host development and aging. Bifidobacterium is a microbial genus found in the human digestive tract that has probiotic activities such as improving constipation and enhancing immunity. The species and numbers present change with age, but there has been limited research on probiotic gut microbiota at specific ages. This study analyzed the distribution of 610 bifidobacteria in subjects in several age groups (0−17, 18−65, and 66−108 y) using 486 fecal samples and determined the distribution of glycoside hydrolases based on genetic analysis of strains representing 85% of the Bifidobacterium species abundance in each age group. 6’-Sialyllactose is a major component of acidic breast milk oligosaccharides, which can promote human neurogenesis and bifidobacteria growth. Using genotypic and phenotypic association analysis, we investigated the utilization of 6’-sialyllactose by six B. bifidum strains isolated from subjects 0–17 and 18–65 y. A comparative genomic analysis of the six B. bifidum strains revealed differences in genomic features across age groups. Finally, the safety of these strains was evaluated by antibiotic gene and drug resistance phenotype analysis. Our results reveal that the distribution of glycoside hydrolase genes in B. bifidum varies with age, thus affecting the phenotypic results. This provides important insights for the design and application of probiotic products for different ages.

Colostrum represents an important source for the transfer of important commensal bacteria from mother to newborn and has a strong impact on the newborn’s health after birth. However, the composition of the colostrum microbiome is highly heterogeneous due to geographic factors and ethnicity (maternal, cultural, and subsistence factors). By analyzing the colostrum 16S rRNA gene full-length sequencing dataset in 97 healthy mothers (60 from Han, 37 from Li) from the Hainan island of China, we showed that the ethnic differences of the colostrum microbiome in a maternal cohort with different ethnic origins shared physical geography. Results indicated that the richness of microbial community in colostrum of Han women was higher than that of Li women, but there was no significant difference in Shannon index and invsimpson index between the two groups. Visualization analysis based on the distance showed an obvious ethnicity-associated structural segregation of colostrum microbiota. The relative abundance of Firmicutes was higher in the microbiota of the Han group than in Li’s, while Proteobacteria was on the contrary. At the genus level, the most dominant members of the Han and Li ethnic groups were Acinetobacter and Cupriavidus, two common environmental bacteria, respectively, although skin-derived Staphylococcus and Streptococcus were still subdominant taxa. Cupriavidus lacunae was the most dominant species in the Li group, accounting for 26.10% of the total bacterial community, but only 3.43% for the Han group with the most dominant Staphylococcus petrasii (25.54%), indicating that human colostrum microbiome was more susceptible to local living environmental factors. Hence, the ethnic origin of individuals may be an important factor to consider in human milk microbiome research and its potential clinical significance during the perinatal period in ethnic-diverse societies, even within a small geographic scale.

Acrylamide, a II A carcinogen, widely exists in fried and baked foods. L-asparaginase can inhibit acrylamide formation in foods, and enzymatic stability is the key to its application. In this study, the Escherichia coli L-asparaginase (ECA) stable variant, D60W/L211R/L310R, was obtained with molecular dynamics (MD) simulation, saturation mutation, and combinatorial mutation, the half-life of which increased to 110 min from 60 min at 50 °C. Furthermore, the working temperature (maintaining the activity above 80%) of mutation expanded from 31 °C–43 °C to 35 °C–55 °C, and the relative activity of mutation increased to 82% from 65% at a pH range of 6–10. On treating 60 U/mL and 100 U/g flour L-asparaginase stable mutant (D60W/L211R/L310R) under uncontrolled temperature and pH, the acrylamide content of potato chips and bread was reduced by 66.9% and 51.7%, which was 27% and 49.9% higher than that of the wild type, respectively. These results demonstrated that the mutation could be of great potential to reduce food acrylamide formation in practical applications.

Purpose Bifidobacterium is an important probiotic used in food and medicine production. The probiotic properties of bifidobacteria are strain specific, so it is necessary to evaluate the probiotic properties of bifidobacteria isolated from specific populations, especially when developing products suitable for specific populations. The objective of this study was to evaluate the probiotic potential and safety of bifidobacteria isolated from healthy Uyghur infants from northwestern China. Methods In this study, antimicrobial activity, antibiotic sensitivity, hemolytic, acid and bile tolerance, hydrophobicity, co-aggregation, auto-aggregation, and antioxidant activity were evaluated. Results Based on antagonistic activity spectrum against seven intestinal pathogenic bacteria, 14 excellent strains were initially selected. Among 14 strains, four bifidobacteria strains (BF17-4, BF52-1, BF87-3, and BF88-5) were superior to strain Lactobacillus rhamnosus GG in cell surface hydrophobicity and auto-aggregation percentages and close to strain GG in co-aggregation with Escherichia coli EPEC O127: K63 (CICC 10411). The antioxidant activities of each of the 14 bifidobacteria strains varied with the cell components. Most of the strains were sensitive to all the antimicrobials tested, except kanamycin and amikacin. Conclusion BF17-4 and BF52-1 are good candidates for further in vivo studies and further used in functional foods.

The maternal gut is thought to be the principal source of potential probiotic bacteria in the infant gut during the lactation stage. It is not clear whether facultative symbiont lactobacilli strictly follow vertical transmission from mother to infant and display the ethnic specificity in terms of species and strain composition in mother–infant cohorts. In the present study, a total of 16 former Lactobacillus species (365 strains) and 11 species (280 strains) were retrieved from 31 healthy mother–infant pairs of two ethnic groups, which have never intermarried, respectively. The result showed that the composition and number of Lactobacillus species between the two ethnic groups varied. Among 106 Lacticaseibacillus paracasei strains isolated, 64 representative strains were classified into 27 sequence types (ST) by means of multilocus sequence typing (MLST), of which 20 STs derived from 33 Uighur strains and 7 STs from 31 Li strains, and no homologous recombination event of genes was detected between strains of different ethnic groups. A go-EBURST analysis revealed that except for a few mother–infant pairs in which more than one STs were detected, L. paracasei isolates from the same mother–infant pair were found to be monophyletic in most cases, confirming vertical transfer of Lactobacillus at the strain level. More notably, L. paracasei isolates from the same ethnic group were more likely than strains from another to be incorporated into a specific phylogenetic clade or clonal complex (CC) with similar metabolic profile of glycan, supporting the hypothesis of ethnic specificity to a large degree. Our study provides evidence for the development of personalized probiotic tailored to very homogenous localized populations from the perspective of maternal and child health.

The maternal gut is thought to be the principal source of potential probiotic bacteria in the infant gut during the lactation stage. It is not clear whether facultative symbiont lactobacilli strictly follow vertical transmission from mother to infant and display the ethnic specificity in terms of species and strain composition in mother-infant cohorts. In the present study, a total of 16 former

Staphylococcus aureus is one of the main pathogens causing mastitis in dairy animals worldwide. It is an important opportunistic pathogen of raw milk, and the enterotoxin causes significant food poisoning. Monitoring the antibiotic resistance of S. aureus in raw milk is helpful for a risk assessment of S. aureus. In this study, 62 strains (43.1%) of S. aureus were isolated from 144 retail raw milk samples of different varieties from four regions in northern Xinjiang, China. Among them, the isolation rates at Shihezi, Hami, Altay, and Tacheng were 58.1% (54/93), 12.9% (4/31), 18.2% (2/11), and 22.2% (2/9), respectively. The isolation rate of positive strains in cow milk samples was the highest (61.7%, 37/60), followed by camel milk (35.9%, 23/64), and horse milk (10.0%, 2/20). The results of the classical virulence genes test showed that 12.9% (8/62) of the isolates carried at least one virulence gene. The main genotype was see (6.5%, 4/62), followed by sea+sec (3.2%, 2/62), sea (1.6%, 1/62), and sec (1.6%, 1/62). The analysis of 13 resistance genes and the susceptibility to 12 different antibiotics of 62 isolates showed that 80.6% (50/62) of the strains were resistant to at least one antibiotic, and 46.8% (29/62) were resistant to three or more antibiotics. The isolated strains had the highest resistance rate to penicillin (72.6%, 45/62), and 25.8% (16/62) of the isolates carried the blaZ resistance gene. In addition, 32 strains (51.6%, 32/62) of methicillin-resistant S. aureus were detected. All isolates had the ability to form biofilms. The pulsed-field gel electrophoresis results showed that the 47 isolates revealed 13 major pulsotypes (P1–P13) and 26 subtypes with 80% similarity, indicating the overall genetic diversity in the distribution area and sources of the samples. These findings indicate that S. aureus causes serious pollution of raw milk in northern Xinjiang, which has a negative effect on public health. Therefore, control measures and continuous monitoring should be undertaken to ensure the quality and safety of raw milk.

Psychrophiles inhabiting various cold environments are regarded as having evolved diverse physiological and molecular strategies, such as the accumulation of trehalose to alleviate cold stress. To investigate the possible contributions of trehalose metabolism-related enzymes to cold-adaption in psychrotrophic bacteria and enrich the resource bank of trehalose hydrolysis enzymes, a novel cold-adapted GH15 GA-like trehalase (MpTre15A) from psychrotolerant Microbacteriumphyllosphaerae LW106 isolated from glacier sediments was cloned and characterized. The recombinant MpTre15A from M. phyllosphaerae LW106 was expressed and purified in Escherichia coli BL21(DE3). The purified MpTre15A functioned as a hexamer and displayed maximal activity at pH 5.0 and 50 °C. Substrate specificity assay proved MpTre15A only showed hydrolytic activity toward α,α-trehalose. Site-directed mutation verified the key catalytic sites of Glu392 and Glu557 in MpTre15A. The kcat and kcat/Km values of MpTre15A at 4 °C (104.50 s−1 and 1.6 s−1 mM−1, respectively) were comparable to those observed for thermophilic GH15 trehalases at 50 °C, revealing its typical cold-adaptability. MpTre15A showed a trehalose conversion rate of 100% and 99.4% after 10 min and 15 min of incubation at 50 °C and 37 °C, respectively. In conclusion, this novel cold-adapted α,α-trehalase MpTre15A showed potential application for developing therapeutic enzymes, enzyme-based biosensors, and enzyme additives in the fermentation industry.

Breast milk acts as an intermediary for the transfer of functionally important commensal bacteria from mother to infant, especially for Bifidobacterium that can colonize the infant gut. However, the vast majority of rRNA amplicon-based studies reported the conspicuous intercohort and interindividual variation for the prevalence of Bifidobacterium in breast milk. In order to elucidate whether Bifidobacterium phylotypes persistently co-occured at the species or strain level in mother–breast milk–infant triads, we analyzed collectively the next-generation sequencing (NGS) datasets of bacterial 16S rRNA gene and the Bifidobacterium-specific groEL gene from maternal feces, breast milk, and infant feces in a small yet very homogeneous cohort of 25 healthy Uyghur mother–infant pairs (lactation for 7–720 days) in Kashgar, Xinjiang, China. Overall, 16S rRNA gene analysis showed that microbiome in the newborn gut was closer to that of breast milk in the first 4 months of lactation, and subsequently showed an obvious trend of adulthood at 6–12 months. Based on the BLAST accurate taxonomic result of the representative sequences of all ASVs (amplicon sequencing variants), only three sets of ASVs could be clearly assigned into Bifidobacterium species, whereas the remaining eight sets of ASVs corresponded to four indefinite Bifidobacterium species group. By contrast, the groEL gene dataset was partitioned into 376 ASVs, at least belonging to 13 well-known Bifidobacterium species or subspecies, of which 15 ASVs, annotated to seven well-known Bifidobacterium species or subspecies, showed triadic synchronism in most 23 mother–infant pairs tested. However, several other rare bifidobacterial phylotypes, which were frequently encountered in animals, were found to display no correspondence of the presence between the three ecosystems of mother–infant pairs. Our test results were obviously to support the hypothesis that breast milk acts as an intermediary for the transfer of probiotic commensal bacteria from mother to infant, especially for endosymbiotic Bifidobacterium that can colonize the infant gut. Some oxygen-insensitive exogenous Bifidobacterium phylotypes with a cosmopolitan lifestyle may be indirectly transferred to breast milk and the infant’s intestinal tract through environmental contamination. Thus, the groEL gene proved to be a very effective target for the depth resolution of Bifidobacterium community by high-throughput sequencing technologies.

Staphylococcus aureus is one of the main pathogens causing mastitis in dairy animals worldwide. It is an important opportunistic pathogen of raw milk, and the enterotoxin causes significant food poisoning. Monitoring the antibiotic resistance of S. aureus in raw milk is helpful for a risk assessment of S. aureus. In this study, 62 strains (43.1%) of S. aureus were isolated from 144 retail raw milk samples of different varieties from four regions in northern Xinjiang, China. Among them, the isolation rates at Shihezi, Hami, Altay, and Tacheng were 58.1% (54/93), 12.9% (4/31), 18.2% (2/11), and 22.2% (2/9), respectively. The isolation rate of positive strains in cow milk samples was the highest (61.7%, 37/60), followed by camel milk (35.9%, 23/64), and horse milk (10.0%, 2/20). The results of the classical virulence genes test showed that 12.9% (8/62) of the isolates carried at least one virulence gene. The main genotype was see (6.5%, 4/62), followed by sea+sec (3.2%, 2/62), sea (1.6%, 1/62), and sec (1.6%, 1/62). The analysis of 13 resistance genes and the susceptibility to 12 different antibiotics of 62 isolates showed that 80.6% (50/62) of the strains were resistant to at least one antibiotic, and 46.8% (29/62) were resistant to three or more antibiotics. The isolated strains had the highest resistance rate to penicillin (72.6%, 45/62), and 25.8% (16/62) of the isolates carried the blaZ resistance gene. In addition, 32 strains (51.6%, 32/62) of methicillin-resistant S. aureus were detected. All isolates had the ability to form biofilms. The pulsed-field gel electrophoresis results showed that the 47 isolates revealed 13 major pulsotypes (P1–P13) and 26 subtypes with 80% similarity, indicating the overall genetic diversity in the distribution area and sources of the samples. These findings indicate that S. aureus causes serious pollution of raw milk in northern Xinjiang, which has a negative effect on public health. Therefore, control measures and continuous monitoring should be undertaken to ensure the quality and safety of raw milk.

In this study, the chemical structure and bioactive properties of the EPS of Pediococcus acidilactici MT41-11 isolated from camel milk were investigated. Two polysaccharide fractions (EPS-1, EPS-2) with molecular weights about 69.0 kDa were obtained, which were purified using DEAE-Sepharose and Sephadex G-100 chromatography. Based on monosaccharide composition, FT-IR, and 1D, 2D NMR spectra, concluded that EPS-1 had a backbone composed of →2)-α- d -Manp-(1→, →3)-α- d -Manp-(1→ and with branches containing α- d -Manp-(1→, EPS-2 had a backbone composed of →6)-β- d -Glcp-(1→, and with branches containing →2)-α- l -Fucp-(1→, →3)-α- d -Glcp-(1→, →2)-α- d -Glcp-(1→, β- d -Glcp-(1→, and α- d -Glcp-(1→. Remarkably, in vitro assays showed that EPS possessed multiple bioactive properties, including stimulating Lactobacillus growth and a high DPPH free radical scavenging activity. Also, it has a good ability to anti-biofilms. Overall, the analysis of all data showed EPS from P. acidilactici MT41–11 can be used as anti-oxidant, anti-biofilm agent, and also as a potential candidate prebiotic for health food or medicine industry.

Elaeagnus angustifolia var. orientalis (L.)Kuntze fruit contains a large number of naturally occurring molecules present as glycoside, methylated, and methyl ester conjugates, which should be hydolysed or transformed to become bioactive forms. For this purpose, Bifidobacterium animalis subsp. lactis HN-3 was selected to ferment Elaeagnus angustifolia var. orientalis (L.)Kuntze fruit juice (EOJ). After fermentation, the total phenolic content (TPC) and antioxidant capacity of the EOJ increased significantly compared to the non-fermented EOJ. Using widely-targeted metabolomics analysis, polyphenolic compounds involved in the flavonoid biosynthetic pathway were determined to be up-regulated in the fermented EOJ. In addition, the metabolites generated by 8 deglycosidation, 5 demethylation, 5 hydrogenation, and 28 other reactions were detected in higher concentrations in the fermented EOJ compared to the non-fermented EOJ. Interestingly, these up-regulated metabolites have higher antioxidant and other biological activities than their metabolic precursors, which provide a theoretical basis for the development of Bifidobacterium-fermented plant products with stronger functional activities.

The maternal gut is the principal source of commensal bacteria in the infant gut during the lactation stage, where breast milk acts as an intermediary for the transfer of potential probiotic bacteria consortia, including Lactobacillus. This study aimed to characterize the bacterial communities in human milk, maternal, and infant feces in a small yet very homogeneous cohort of 25 healthy mother–infant pairs in northwestern China (n = 25, infant age from 7 days to 2 years), with special emphasis on the cooccurrence and vertical transfer of Lactobacillus phylotypes at the species or strain level in mother-breast milk-infant triads. Accurate sequencing analysis revealed that among 73 Lactobacillus zero-radius operational classification units (ZOTUs) identified, 58 belonging to 18 recognized species or species groups were distributed in all three types of samples. Lactobacillus ruminis, L. mucosae and L. gasseri-johnsonii as true residents were the most represented in all three ecosystems, whereas the content of Lactobacillus phylotypes commonly developed as probiotics was not dominant. While the numbers of Lactobacillus species in breast milk and infant feces were greater than that in maternal feces, principal coordinates analysis (PCoA) based on beta diversity, coupled with the frequency of isolates determined by culture methods, showed that the Lactobacillus community in the infant gut was more similar to that in the maternal gut than to that in breast milk, suggesting that the gut is niche selective for Lactobacillus populations. In addition, identical strains of L. ruminis, L. paracasei, L. mucosae and L. salivarius were isolated from multiple mother–infant pairs, supporting the hypothesis that vertical transfer of bacteria via breastfeeding contributes to the initial establishment of the microbiota in the developing infant intestine.

The present work reported the complete genome sequence analysis of Acidithiobacillus ferrivorans strain XJFY6S-08 isolated from acid mine drainage in Fuyun copper mine in Xinjiang, China, revealing the potential for extreme environmental adaptation. The strain XJFY6S-08 possesses 3,161,380 bp in length and 56.55% GC content. Genomic analysis revealed that this strain harbors metal-tolerant genes coding for the mer operon, arsRBC operon and a variety of metal assimilation and efflux proteins. Genes coding for K+/H+ transporting ATPase and the Na+/H+ antiporter gene nhaA for pH adaptation were identified. The presence of genes associated with various DNA repair enzymes and the synthesis of mycosporine-like amino acids precursor support the UVR resistance mechanisms. The genes related to membrane modifications (ppiBCD, slyD, surA, cfa and fabF) and resistance–nodulation–division (RND) family can play a crucial role in organic solvents tolerance. The strain XJFY6S-08 resists low-temperature conditions by a set of mechanisms such as changes of RNA metabolism, transmembrane transport, compatible solutes and transport, biofilm and EPS formation, chemotaxis and motility and ROS scavenging. These findings can provide important information for further studying the comparative genome and environmental adaptation mechanism of A. ferrivorans.

In the present study 114 lactic acid bacteria strains, isolated from raw mare milks from pastoral areas for ethnic minorities in northwest China, were screened for probiotic traits, and their characteristics were compared with those of Lactobacillus rhamnosus GG, a commercial strain. Among the 114 strains identified, the most common species was Pediococcus pentosaceus (n = 52), followed by Leuconostoc lactis (n = 35), Lactobacillus helveticus (n = 7), Lactobacillus plantarum (n = 6), Lactobacillus kefiri (n = 5), Lactobacillus curvatus (n = 4), Lactobacillus paracasei (n = 3), and Lactococcus garvieae (n = 3). Based on acid and bile salt tolerance, 15 strains were further selected. All selected strains were subjected to a series of in vitro tests to assess their technological properties, including cell surface hydrophobicity (13.6-56.2%), autoaggregation ability (9.26-38.30%), coaggregation ability, and heat and lysozyme survival rates (84.74-94.01% and 80.52-99.37%, respectively). In vitro antagonism showed that Lb. plantarum (M5-19, M8-60, M8-59) exhibited the most strong inhibitory activity against 7 tested pathogens. Moreover, antibiotic resistance and hemolytic activity were investigated for safety assessment. No strain exhibited hemolytic activity, and most of the strains were sensitive to a series of 14 antibiotics of clinical importance. Ultimately, the principal component analysis of all data obtained above showed that 2 Lb. plantarum strains (M8-59, M8-60) and Lb. paracasei M1-36 exhibited the best potential for their inclusion as adjunct functional cultures in local fermented dairy products.

Background Bifidobacteria are important probiotics; some of the beneficial effects of bifidobacteria are achieved by the hydrolysis of glycans in the human gut. However, because the diet of breastfed infants typically lacks plant-derived glycans, in the gut environment of mothers and their breastfed infants, the mother will intake a variety of plant-derived glycans, such as from onions and bananas, through her diet. Under this assumption, we are interested in whether the same species of bifidobacteria isolated from mother-infant pairs present a distinction in their hydrolysis of plant-derived carbohydrates. Results Among the 36 Bifidobacterium strains, bifidobacterial carbohydrate utilization showed two trends related to the intestinal environment where the bacteria lived. Compared with infant-type bifidobacterial strains, adult-type bifidobacterial strains preferred to use plant-derived glycans. Of these strains, 10 isolates, 2 Bifidobacterium pseudocatenulatum (B. pseudocatenulatum), 2 Bifidobacterium pseudolongum (B. pseudolongum), 2 Bifidobacterium bifidum (B. bifidum), 2 Bifidobacterium breve (B. breve), and 2 Bifidobacterium longum (B. longum), were shared between the mother-infant pairs. Moreover, the repetitive sequence-based polymerase chain reaction (rep-PCR) results illustrated that B. pseudolongum and B. bifidum showed genotypic similarities of 95.3 and 98.2%, respectively. Combined with the carbohydrate fermentation study, these results indicated that the adult-type strains have a stronger ability to use plant-derived glycans than infant-type strains. Our work suggests that bifidobacterial carbohydrate metabolism differences resulted in the selective adaptation to the distinct intestinal environment of an adult or breastfed infant. Conclusions The present study revealed that the different gut environments can lead to the differences in the polysaccharide utilization in the same strains of bifidobacterial strains, suggesting a further goal of investigating the exact expression of certain enzymes in response to specific carbon sources.

BackgroundAs a secure vehicle of probiotics, breast milk can provide babies with the Lactobacillus which can not only colonize and inhibit the pathogenic bacterial infection in infant's intestines. The purpose of this study was to assess the occurrence and probiotic properties of Lactobacillus spp. of maternal breast milk in the Uyghur population, Xinjiang China, by using culture method. Results Based on repetitive genomic fingerprinting PCR (rep-PCR), a total of 198 isolates of L actobacill us from 31 different lactation of breast milk samples of Uighur in kashi region of Xinjiang were classified into 11 genotypes, which were identified as Lactobacillus fermentum (82 isolates, prevalence: 61.3%, mean relative abundance: 45.4%, genotype: 3), Lactobacillus brevis (75, 41.9%, 38.1%, 4), Lactobacillus oris (37, 32.3%, 15.1%, 3) and rare Lactobacillus vaginalis (4, 9.7%, 1.4%, 1). From colostrum to mature milk, the number and species of Lactobacillus showed an uptrend, from mature milk to late milk, a downside was found in Lactobacillus. The relative abundance of L. fermentum decreased throughout lactation, while L . brevis showed an opposite trend. Three isolates were randomly selected for each genotype to estimate antimicrobial activity (33 isolates in total). Among the isolates, 20 isolates exhibited broad antibacterial spectrum with inhibition halos >10 mm against most indicator pathogens, which were selected for assessing probiotic properties. Nineteen isolates showed the resistance to vancomycin and co-trimoxazole, and KM66 ( L . brevis ) was resistant up to 5/8 antibiotics. KM147 ( L. brevis ) and Y3 ( L. fermentum ) strains were selected as potential probiotics by combining lysozyme, acid and bile salt tolerance. Conclusion Species composition and prevalence of Lactobacillus varied with the study subjects throughout lactation phase. The deliberately selected Lactobacillus strains from breast milk may have a great potential as probiotics to inhibit pathogen infection in infants.

Additional file 2.

The aim of the present study was to determine the repairing effects of intercellular adhesion molecule (ICAM)-1-expressing mesenchymal stem cells (MSCs) in mice with autoimmune thyroiditis. Following induction of an experimental autoimmune thyroiditis (EAT) model, mice were randomly divided into the following groups (n=10 each): i) Normal control; and experimental groups that were subject to EAT induction, including ii) EAT model; and iii) primary MSC; iv) C3H10T1/2/MSC; v) C3H10T1/2-MIGR1/MSC; and vi) C3H10T1/2-MIGR1-ICAM-1/MSC, which were all administered the relevant cells. MSCs were administered via the caudal vein. A blood sample was harvested from the angular vein of each animal 28 days post-treatment and ELISA was used to determine the serum total triiodothyronine, total thyroxine (T4), thyroid-stimulating hormone (TSH), anti-thyroid peroxidase (TPOAb), anti-thyroid microsomal (TMAb) and anti-thyroglobulin (TGAb) antibodies. Hematoxylin and eosin staining was performed to evaluate injury of the thyroid gland by determining the size of the follicle, inflammatory infiltration, colloidal substance retention and epithelial injury. Reverse transcription-quantitative polymerase chain reaction was performed to determine the mRNA expression of interleukin (IL)-4, IL-10, IL-17 and interferon (INF)-γ. Western blot analysis was performed to determine the expression of p38 mitogen-activated protein kinase (p38) and extracellular signal-regulated kinase (ERK). To observe cellular migration in vivo, mice were divided into the following groups, (n=10 each), which were subject to EAT induction: i) CM-DiI-labeled primary MSC; ii) CM-DiI-labeled C3H10T1/2/MSC; iii) CM-DiI-labeled C3H10T1/2-MIGR1/MSC; and iv) CM-DiI-labeled C3H10T1/2-ICAM-1/MSC, which were all administered the relevant cells via the caudal vein. C3H10T1/2-ICAM-1/MSCs were able to ameliorate the expression of T4, TSH, TPOAb, TMAb and TGAb in vivo, attenuate thyroid follicle injury and decrease the splenic index in mice. They were also able to ameliorate the mRNA expression of IL-4, IL-10, IL-17 and INF-γ, and the modulation of the P38 and ERK-signaling pathways in the mouse spleen. Furthermore, ICAM-1 overexpression was able to modulate the nesting of MSCs in the thyroid gland and lung. These findings suggest that C3H10T1/2-ICAM-1/MSC may affect the differentiation, proliferation and migration of immunocytes through modulating the p38 and ERK signaling pathways, and that ICAM-1 may modulate the immunoregulatory effects of MSCs by affecting the migration of MSCs in vivo.

Phomopsis sp. XP-8, an endophytic fungus from the bark of Tu-Chung (Eucommia ulmoides Oliv) showed capability to biosynthesize pinoresinol (Pin) and pinoresinol diglucoside (PDG) from glucose (glu) and phenylalanine (Phe). To verify the mass flow in the biosynthesis pathway, [13C6]-labeled glu and [13C6]-labeled Phe were separately fed to the strain as sole substrates and [13C6]-labeled products were detected by ultra-high-performance liquid chromatography-quadrupole time of flight mass spectrometry. As results, [13C6]-labeled Phe was incorporated into [13C6]-cinnamylic acid (Ca) and p-coumaric acid (p-Co), and [13C12]-labeled Pin, which revealed that the Pin benzene ring came from Phe via the phenylpropane pathway. [13C6]-Labeled Ca and p-Co, [13C12]-labeled Pin, [13C18]-labeled pinoresinol monoglucoside (PMG), and [13C18]-labeled PDG products were found when [13C6]-labeled glu was used, demonstrating that the benzene ring and glucoside of PDG originated from glu. It was also determined that PMG was not the direct precursor of PDG in the biosynthetic pathway. The study identified the occurrence of phenylalanine- lignan biosynthesis pathway in fungi at the level of mass flow.

Acidithiobacillus ferrooxidans YNTRS-40 (A. ferrooxidans) is a chemolithoautotrophic aerobic bacterium isolated from Tengchong hot springs, Yunnan Province, China, with a broad growth pH range of 1.0–4.5. This study reports the genome sequence of this strain and the information of genes related to the adaptation of diverse stresses and the oxidation of ferrous iron and sulfur. Results showed that YNTRS-40 possesses chromosomal DNA (3,209,933-bp) and plasmid DNA (47,104-bp). The complete genome of 3,257,037-bp consists of 3,349 CDS genes comprising 6 rRNAs, 52 tRNAs, and 6 ncRNAs. There are many encoded genes associated with diverse stresses adaptation and ferrous iron and sulfur oxidation such as rus operon, res operon, petI, petII, sqr, doxDA, cydAB, and cyoABCD. This work will provide essential information for further application of A. ferrooxidans YNTRS-40 in industry.

Acidithiobacillus ferrooxidans YNTRS-40 ( A. ferrooxidans ) is a chemolithoautotrophic aerobic bacterium isolated from Tengchong, Yunnan Province, China and with a broad growth pH range of 1.0-4.5. This study reported the genome sequence of this strain and the information of genes related to the adaptation of diverse stresses and the oxidation of ferrous iron and sulfur. Results showed that YNTRS-40 possesses chromosomal DNA (3,209,933-bp) and plasmid DNA (47,104-bp). The complete genome of 3,257,037-bp was consist of 3349 CDS genes comprising 6 rRNAs, 52 tRNAs and 6 ncRNAs. There are many encoded genes associated with diverse stresses adaptation and ferrous iron and sulfur oxidation such as rus operon, res operon, pet Ⅰ , pet Ⅱ , sqr gene, doxDA , cydAB and cyoABCD . This study will provide essential information for further application of A. ferrooxidans YNTRS-40 in industry.

Background There are still a large variety of microorganisms among aquatic animals, especially probiotic lactic acid bacteria in cold water fishes at high latitudes have not been fully developed. Hence, the present study aims to evaluate the probiotic potential of cold-adapted Lactobacillus strains isolated from the intestinal tract of cold water fishes (Xinjiang) and select candidates to be used as new food preservative agents and/or probiotic additives in feeding of aquaculture. Results A total of 43 Lactobacillus spp. were isolated from 16 kinds of intestinal tract of cold-water fishes. They were characterized by phenotypic methods, identified using Rep-PCR and 16S rRNA gene sequencing as four species: Lactobacillus sakei (22 isolates), Lactobacillus plantarum (16 isolates), Lactobacillus casei (4 isolates) and Lactobacillus paracasei (1 isolate). The in vitro tests included survival in low pH and bile, antimicrobial activity (against Escherichia coli, Salmonella enterica subsp. enterica serovar Typhimurium, Salmonella enterica subsp. enterica, Listeria monocytogenes, and Listeria innocua), resistance to 15 antibiotics and hemolytic tests. Among all 43 lactobacilli isolates, the 22 isolates showed a wide range of antimicrobial activity against 6 different pathogenic strains. There were twenty isolates growing at optimal temperature ranging 16~20 °C, which were initially considered to be cold-adapted strains. Two (2) Lb. sakei strains and 2 Lb. plantarum strains demonstrated the highest survivability after 4 h of exposure at pH 2.0. Most of the tested strains cannot be cultured after exposed into 0.5% bile solution for 4 h, while 2 Lb. plantarum strains (E-HLM-3, CQ-CGC-2) and 1 Lb. sakei strain M-DGM-2 survived even at 2% bile concentration. In addition, the safety assessment showed that 22 strains without any detectable hemolytic activity and resistant to glycopeptides (vancomycin, teicoplanin), levofloxacin, aztreonam, amikacin and oxacillin, while all the studied lactobacilli showed sensitivity to or semi-tolerant to other antibiotics. Conclusions Based on all the experiments, 3 strains, including E-HLM-3, CQ-CGC-2, and M-DGM-2 might be a candidate of choice for using in the food preservative agents and/or probiotic additives in feeding of aquaculture.

Four kinds of oligosaccharides were used as co-encapsulating agents to test the effect of extrusion-based microencapsulation on protection of Lactobacillus fermentum L7 against exposure to simulated gastric and intestinal juices as well as long-term refrigeration storage at 4 °C. The combination of alginate with galacto-oligosaccharides, isomalto-oligosaccharides, fructo-oligosaccharides, and xylo-oligosaccharides, or alginate alone exhibited good properties of the beads. The diameters of the cell beads co-encapsulated with oligosaccharides and encapsulated with alginate alone were similar, in the range of 2.34–2.51 mm. However, the encapsulation yield of L. fermentum cells co-encapsulated with oligosaccharides, which was in the range of 79.52–89.75%, was significantly higher than that of cells encapsulated with alginate alone. The capsules were stable in gastric conditions and can disintegrated when exposed to intestinal conditions. Additionally, the viability of microencapsulated cells after exposure to the simulated gastric and intestinal juices as well as long-term refrigeration storage was better than that of free cells, and the viability of cells co-encapsulated with oligosaccharides was better than that of cells encapsulated with alginate alone. Furthermore, fructo-oligosaccharides used as co-encapsulating agent showed the best performance. Microencapsulating L. fermentum with oligosaccharides protected cells well at a low temperature and offered effective gastrointestinal delivery of probiotics, and thus has the potential to maintain bacterial survival in probiotic products and will provide the research basis for design of effective probiotic-prebiotic combinations to maximize host benefit.

Asian cold-based glacier yeasts with respect to their abundance, distribution, and taxonomy, in contrast to other continental cryosphere areas, have been little investigated. The present study reports the diversity and phylogeny of culturable cold-adapted yeasts in six cold habitats of the Glacier No.1 in the Tianshan Mountains (northwestern China). Of the total 591 yeast isolates, 401 were identified as basidiomycetous yeasts represented by 41 species of 15 genera, while 190 ascomycetous yeast isolates were assigned to the 8 species of 7 genera. The most prevalent species was Candida akabanensis with a 19.8% frequency of occurrence, followed by Vishniacozyma victoriae (16.4%) and Diutina rugosa (9.98%), of which V. victoriae was the only yeast species common to all six glacial habitats. Variability on the component and abundance of yeast taxa among glacial habitats primarily displayed in four dominant yeast genera, namely Candida, Vishniacozyma, Filobasidium, and Naganishia. However, network analysis illustrated that most of 32 rare yeast populations were habitat-specific, implying that the low-abundance yeast population was more easily influenced by the local habitat conditions (species sorting). Based on indicator species analyses, the subglacial habitat was characterized by psychrotolerant and/or psychrophilic yeast taxa.

To study the phylogenetic and genetic heterogeneity of 23 Acidithiobacillus strains from various geographical locations, as well as the relationship between the DNA fingerprinting classification and geographical origin of Acidithiobacillus.Partial 16S-23S rRNA gene intergenic spacer (ITS) was used to construct corresponding phylogenetic trees based on the sequence homology. rus gene amplification and rep-PCR assay with two different primers (BOXAIR and ERIC) were performed to analyze genetic heterogeneity of Acidithiobacillus strains from diverse environment.Acidithiobacillus revealed a great genetic heterogeneity. The whole isolates were classified into five groups by ITS sequence analysis. This result was similar with that obtained by rep-PCR. Acidithiobacillus ferrooxidans strains were always divided into two groups of phylogenetic and BOXAIR fingerprinting cluster analysis. However, these were clustered one group in the ERIC dendrogram. Genotypic analysis of the rus gene suggested that different iron oxidation pathways have been evolved in these closely related bacteria. Taken together, the iron oxidation pathway of Acidithiobacillus and phylogenetic groups have no obvious correlation. ITS gene has been proven very useful in distinguishing closely related species or subspecies of Acidithiobacillus, to BOXAIR-PCR, which has been recommended as reliable tool for genetic heterogeneity analysis of Acidithiobacillus.

The purpose of the study was to reveal geographic region-related Acidithiobacillus spp. distribution and allopatric speciation. Phylogenetic and diversity analysis was done to expand our knowledge on microbial phylogeography, diversity-maintaining mechanisms and molecular biogeography.We amplified 16S rRNA gene and RubisCO genes to construct corresponding phylogenetic trees based on the sequence homology and analyzed genetic diversity of Acidithiobacillus spp..Thirty-five strains were isolated from three different regions in China (Yunnan, Hubei, Xinjiang). The whole isolates were classified into five groups. Four strains were identified as A. ferrivorans, six as A. ferridurans, YNTR4-15 Leptspirillum ferrooxidans and HBDY3-31 as Leptospirillum ferrodiazotrophum. The remaining strains were identified as A. ferrooxidans. Analysis of cbbL and cbbM genes sequences of representative 26 strains indicated that cbbL gene of 19 were two copies (cbbL1 and cbbL2) and 7 possessed only cbbL1. cbbM gene was single copy. In nucleotide-based trees, cbbL1 gene sequences of strains were separated into three sequence types, and the cbbL2 was similar to cbbL1 with three types. Codon bias of RubisCO genes was not obvious in Acidithiobacillus spp..Strains isolated from three different regions in China indicated a great genetic diversity in Acidithiobacillus spp. and their 16S rRNA/RubisCO genes sequence was of significant difference. Phylogenetic tree based on 16S rRNA genes and RubisCO genes was different in Acidithiobacillus spp..

Kazak artisanal cheese is one of the famous fermented food in Uighur Autonomy Region of Xinjiang, China. However, the microbial ecology in Kazak artisanal cheeses across different regions is unclear. In this study, we determined the microbial community composition through amplicon sequencing and measured the flavor profile of 10 cheese samples from different regions of Xinjiang. The associations between microbial communities, flavors and environmental factors were examined by redundancy analysis and Monte Carlo permutation test. Cheeses from different regions had different microbial communities, which was mainly reflected in the relative abundance of Lactobacillus, Streptococcus, Issatchenkia, Debaryomyces and Kluyveromyces. In addition, Pichia and Torulaspora were also the key microbial groups, according to the high relative abundance and large co-occurrence incidence in the correlation network. Using the microbe-metabolites correlation analysis, the major flavor-producing taxa were identified as Kluyveromyces, Anoxybacillus, Torulaspora, Lactobacillus, Streptococcus and Dipodascus. Environmental factors accounted for the majority of the microbial community variations, 88.54% for bacteria and 75.71% for fungi. Compared to physico-chemical factors (temperature, moisture, and pH), geographical factors (longitude, latitude and elevation) had a stronger effect on microbial communities in cheese samples from different regions of Xinjiang.

Acidithiobacillus ferrooxidans YNTRS-40 (A. ferrooxidans) is a chemolithoautotrophic aerobic bacterium isolated from Tengchong hot springs, Yunnan Province, China, with a broad growth pH range of 1.0–4.5. This study reports the genome sequence of this strain and the information of genes related to the adaptation of diverse stresses and the oxidation of ferrous iron and sulfur. Results showed that YNTRS-40 possesses chromosomal DNA (3,209,933-bp) and plasmid DNA (47,104-bp). The complete genome of 3,257,037-bp consists of 3,349 CDS genes comprising 6 rRNAs, 52 tRNAs, and 6 ncRNAs. There are many encoded genes associated with diverse stresses adaptation and ferrous iron and sulfur oxidation such as rus operon, res operon, petI, petII, sqr, doxDA, cydAB, and cyoABCD. This work will provide essential information for further application of A. ferrooxidans YNTRS-40 in industry.

The aim of this paper is to determine the efficiency of bioleaching of arsenic in realgar, a Chinese mineral drug, using pure cultures of Acidithiobacillus ferrooxidans or Acidithiobacillus thiooxidans and a mixed culture of A. ferrooxidans and A. thiooxidans. The experiments were carried out in shaker flasks, at 150 rpm, 30 °C at a culture pH of 1.80. To investigate the mechanism of the bioleaching in realgar, media with and without ferrous iron were chosen for the experiments. The results showed that the leaching rate of arsenic in realgar after 20 days was higher (43%) in A. ferrooxidans cultures with ferrous iron compared to cultures without ferrous iron (10%), and the leaching rate of A. thiooxidans cultures only increased from 21% to 23% in the presence of ferrous iron. The leaching rate of arsenic in mixed culture with ferrous iron was greatly enhanced from 16% to 56%, indicating that bioleaching in mixed culture is preferable for the dissolution of realgar.

Methods C57BL/6 mice were treated with porcine thyroglobulin and Freund’s adjuvant both in the simple model group and the BM-MSCs treated group. Mice in the BM-MSCs treated group were injected with homology BM-MSCs (3×10/mouse) at the beginning of model establishment. A normal control group was set as well. All mice were killed at the 28th day after primary immunity. The histopathological analysis of thyroid tissues were observed. The levels of autoantibodies, thyroid hormone, IFN-r, IL-10 were detected.

The purpose of this study is to characterize the community composition and phylogenetic analysis of cyanobacteria from supraglacial cryoconite of the Glacier No. 1 in the Tianshan Mountains, China.We amplified 16S rRNA genes from the extracted cryoconite DNA by PCR with 2 pairs of cyanobacteria-specific primers. Amplificon was used to construct 16S rRNA genes clone library. The estimation of species richness, diversity indices, and rarefaction curve of the 16S rRNA genes library were determined based on representative phylotypes (OTUs).Analysis of 16S rRNA gene sequences allowed grouping of 101 clones into 12 phylotypes (OTUs) using a cut-off of 97% identity. The phylogenetic analysis revealed that most of sequences affiliated to the order Oscillatoriales and Chroococcales except that three were unclassified. The clone library was dominated by representatives of the order Oscillatoriales (81% of the total clones), and the most abundant organisms within this order were in the genus Phormidium (68 clones) including clones grouping into four phylotypes. The only clone of Chroococcales was closely related to the genus Chamaesiphon with 97% similarity. In addition, comparison of soil chemical properties between different habitats indicated that supraglacial cryoconite supported significantly higher the content of available phosphorus and potassium, nitrate nitrogen and organic matter compared with the forefield of the Glacier No. 1.The diversity index of cyanobacteria were relatively high in supraglacial cryoconite of the Glacier No. 1 in the Tianshan Mountains. The community structure was dominated by members of the genus Phormidium. This study may enrich our knowledge on biogeochemical processes and ecological distribution of cyanobacterial populations in glacial ecosystem.

Cheese is a typical handcrafted fermented food in Kazak minority from the Uighur Autonomy Region in China and Central Asia. Among the microbial community that is responsible for Kazak cheese fermentation, yeasts play important role in flavor formation during ripening. To develop ripening cultures, we isolated 123 yeasts from 25 cheese products in Kazak, and identified 87 isolates by the D1/D2 domain of the large subunit rRNA gene sequence. Pichia kudriavzevii was the dominant yeast in Kazak cheese, followed by Kluyveromyces marxianus and Kluyveromyces lactis. Of these, the ability to exhibit enzyme of dominant isolates and contribution to the typical flavor of cheeses was assessed. Enzyme producing yeast strains were inoculated in Hazak cheese‐like medium and volatile compounds were identified by head space solid phase micro extraction coupled to gas chromatography and mass spectroscopy. Pichia kudriavzevii N‐X displayed the strongest extracellular proteolytic and activity on skim milk agar and produced a range of aroma compounds (ethanol, ethyl acetate, 3‐methylbutanol, and acetic acid) for Kazak cheese flavor, could be explored as ripening cultures in commercial production of Kazak cheeses.

Leptospirillum sp. YQP-1, a member of iron-oxidizing bacteria was isolated from volcanic lake in northeast China. Here, we report the draft genome sequence of the strain YQP-1 with a total genome size of 3,103,789bp from 85 scaffolds (104 contigs) with 58.64% G+C content. The genome sequence can be accessed at DDBJ/EMBL/GenBank under the accession no. LIEB00000000.

A monoclonal antibody (MAb) 6H7 raised specifically against granulocytes of scallop (Chlamys farreri) was employed to observe granulocyte occurrence successively in blastulae, gastrulae, trochophore larvae, D-shape larvae, umbo-veliger larvae and creeping larvae of C. farreri by immunohistochemistry assay contrasted with H&E stain using semi-thin sections. Moreover, the reactivity of the MAb with granulocytes of C. farreri, Bay scallop Argopecten irradians, Japanese scallop Patinopecten yessoensis, Blue mussel Mytilus edulis, Pacific oyster Crassostrea gigas and Manila clam Ruditapes philippinarum, was detected by immunofluorescence assay (IFA) with differential interference contrast and fluorescent microscopy and flow cytometric immunofluorescence assay (FCIFA). The results showed that positive signals were first observed at D-shape larval stage, about 28 h post fertilization, after that, umbo-veliger larvae exhibited the positive cells with a diameter of 3–5 μm distributed in velum, digestive gland and esophagus. Then in creeping larvae, the number of positive cells increased with average diameter of 5–7 μm, and widely distributed in foot, digestive gland, gills and adductor muscles. No positive signal was found in blastulae, gastrulae and trochophore larvae. The results of IFA and FCIFA showed MAb 6H7 reacted to granulocytes of C. farreri, A. irradians, P. yessoensis and C. gigas, and the positive percentage reactivity were 53 ± 2.5%, 15 ± 2.5%, 12 ± 2.1% and 19 ± 2.1%, respectively, however, no cross-reaction was detected in hemocytes of R. philippinarum and M. edulis.

We characterized proteolytic bacteria isolates from sediments of the bottom layer of the Glacier No. 1 in the Tianshan Mountains, China. Physiological test and phylogenetic analysis were undertaken to expand our knowledge on diversity and ecological distribution of psycrotrophic and psycrophlic bacteria populations.Using the screening media containing skim milk, we screened cold-adapted strains producing protease. Taxonomic identity and genetic variability of strains isolated was determined by partial 16S rRNA gene sequences and repetitive-element PCR fingerprint.Of the total 125 cold-adapted bacterial isolates, high levels of protease activity were observed from 27 isolates at optimal growth temperatures ranging from 15 to 24 degrees C in plate assay. Among 27 protease-producing strains, only 6 isolates were psychrophilic. The 16S rRNA gene phylogenetic analysis revealed that protease-producing isolates belonged to 5 phyum, namely alpha, beta and gamma of Proteobacteria, Actinobacteria and Cytophaga-Flexibacter-Bacteroides. They are affiliated to the genera Pseudomonas, Polaromonas, Brevundimonas, Rholococces, Cryobacterium, Kocuria, Arthrobacter, Chryseobacterium and Flavobacterium. The populations of the predominant cultivated protease-producing bacteria are the Pseudomonas spp. (40.7%).The results enriched our knowledge on the phylogenetic and physiological diversity of cold-adapted strains producing protease in cold environments.

The purpose of this study was to select cold-adapted lactic acid bacteria (LAB) from the intestinal tract of cold-water fishes from the Eerqisi river Alatai, Xinjiang.By using culture medium MRS and Elliker, isolation of lactic acid bacteria (LAB) from 9 intestinal canal of cold-water fishes was carried out. Taxonomic identity and genetic diversity of strains isolated were determined by partial 16S rRNA gene sequences and rep-PCR with three different kinds of primers named BOX, (GTG)5, ERIC.A total of 78 psychrotrophic isolates were obtained. Among them, 24 isolates had characteristics of LAB and showed the optimal growth temperature ranging from 15 to 24 degrees C. The phylogenesis result showed that 24 strains belonged to 6 Genuses, Carnobacterium (3 strains), Lactococcus (9 strains), Enterococcus (7 strains), Brochothrix (1 strain), Weissella (2 strains), Streptococcus (2 strains), rep-PCR clustering analysis showed that Lactococcus and Enterococcus strains were from different Species. Lactococcus were belonged to 4 species while Enterococcus assigned to 2 species.The phylogenetic diversity of cold-adapted LAB from the intestinal tract of cold-water fishes from the Eerqisi river in the Alatai Xinjiang was relatively abundant.

The purpose of this research is to isolate cold-adapted bacteria producing beta-galactosidase from permafrost sediments of the bottom layer of the Glacier No. 1 in the Tianshan Mountains, China. Physiological test and phylogenetic analysis were undertaken to expand our knowledge on diversity of psycrotrophic and psycrophlic bacteria.By using lactose as the main carbon source and X-Gal as chromogenic agent in the medium, cold-adapted strains producing beta-galactosidase were detected. Taxonomic identity and genetic diversity of strains isolated were determined by spatial 16S rRNA gene sequences and rep-PCR fingerprint. In addition, we analyzed the phonotypic differences between strains showing high similarity of 16S rRNA gene sequences, including the optimum growth temperature, salt tolerance, ability to use carbon source and antibiotic resistance spectra.Of the total 90 cold-adapted bacterial strains isolated, we found 25 stains with beta-galactosidase activity, 76% of which were Gram-positive bacteria. According to growth temperature range, 80% of strains producing beta-galactosidase were identified as psychrophilic bacteria, 20% as psychrotrophs. Phylogeneticlly, the beta-galactosidase-producing bacterial isolates fell in four groups: subclasses alpha, beta and gamma of Proteobacteria, and Firmicutes phylum.The results enrich our knowledge on the phylogenetic and physiological diversity of cold-adapted strains producing beta-galactosidase in cold environments.

The diversity of culturable lipase-producing bacterial strains from permafrost soils at the terminus of a glacier in the Tianshan Mountains was investigated. Isolation and molecular phylogenetic analysis were performed to expand our knowledge on diversity of psychrotrophic and psychrophilic bacteria. In addition, efforts were made focusing on screening for cold active lipases.Lipase-producing bacterial strains were detected on tween 80 and olive oil plates, respectively. Identity and genetic diversity of strains isolated were determined by spatial 16S rRNA gene sequences and rep-PCR fingerprint. The physiological tests were carried out to determine the phonotypic differences between strains showing high similarity of 16S rRNA gene sequences.Of the total 17 bacterial stains exhibiting cold-adapted lipase activity, we found that only 8 stains were able to hydrolyze olive oil. Based on 16S rRNA gene sequences, the lipase-producing bacterial isolates fell in five phylogenetic groups: subclasses (, ( and ( of Proteobacteria, Actinobacteria, and the Cytophaga-Flexibacter-Bacteroides (CFB) phylum. Nearly 59% of the isolates were affiliated with the genus Pseudomonas.The results enrich our knowledge on the psychrotrophic bacterial diversity and biogeographic distribution of cold active lipases-producing bacteria in cold environments.

With the growth of decentralized network users, preserving privacy becomes a critical issue in this open community. Kad-based network, as a typical decentralized system, has been widely used nowadays. However, there is not enough research to achieve anonymity on it. In this paper, we propose an anonymous protocol based on Kad network, named Anonymous Kad (A-Kad), which achieves complete privacy and security for file providers and requesters. A-Kad has the desired property of anonymity and still keeps high efficiency in publishing and querying phases. To achieve anonymity, we establish two anonymous channels which help file providers to anonymously publish file information and securely transfer files. Through these two channels, the file requester can also efficiently query and retrieve files without worrying about exposing its behavior. Moreover, we propose an anonymity degree evaluation model (ADEM) according to three different attacking capabilities and anonymity degree.