Cellulose-degrading bacteria improve conversion efficiency in the co-digestion of dairy and chicken manure by black soldier fly larvae.

Black soldier fly larvae (BSFL) have potential utility in converting livestock manure into larval biomass as a protein source for livestock feed. However, BSFL have limited ability to convert dairy manure (DM) rich in lignocellulose. Our previous research demonstrated that feeding BSFL with mixtures of 40% dairy manure and 60% chicken manure (DM40) provides a novel strategy for significantly improving their efficiency in converting DM. However, the mechanisms underlying the efficient conversion of DM40 by BSFL are unclear. In this study, we conducted a holistic study on the taxonomic stucture and potential functions of microbiota in the larval gut and manure during the DM and DM40 conversion by BSFL, as well as the effects of BSFL on cellulosic biodegradation and biomass production. Results showed that BSFL can consume cellulose and other nutrients more effectively and harvest more biomass in a shorter conversion cycle in the DM40 system. The larval gut in the DM40 system yielded a higher microbiota complexity. Bacillus and Amphibacillus in the BSFL gut were strongly correlated with the larval cellulose degradation capacity. Furthermore, in vitro screening results for culturable cellulolytic microbes from the larval guts showed that the DM40 system isolated more cellulolytic microbes. A key bacterial strain (DM40L-LB110; Bacillus subtilis) with high cellulase activity from the larval gut of DM40 was validated for potential industrial applications. Therefore, mixing an appropriate proportion of chicken manure into DM increased the abundance of intestinal bacteria (Bacillus and Amphibacillus) producing cellulase and improved the digestion ability (particularly cellulose degradation) of BSFL to cellulose-rich manure through changes in microbial communities composition in intestine. This study reveals the microecological mechanisms underlying the high-efficiency conversion of cellulose-rich manure by BSFL and provide potential applications for the large-scale cellulose-rich wastes conversion by intestinal microbes combined with BSFL.

Enhanced protein degradation by black soldier fly larvae (Hermetia illucens L.) and its gut microbes.

Black soldier fly larvae (BSFL) can convert a variety of organic wastes into biomass, and its gut microbiota are involved in this process. However, the role of gut microbes in the nutrient metabolism of BSFL is unclear. In this study, germ-free BSFL (GF) and gnotobiotic BSFL (GB) were evaluated in a high-protein artificial diet model. We used 16S rDNA sequencing, ITS1 sequencing, and network analysis to study gut microbiota in BSFL that degrade proteins. The protein reduction rate of the GB BSFL group was significantly higher (increased by 73.44%) than that of the GF BSFL group. The activity of gut proteinases, such as trypsin and peptidase, in the GB group was significantly higher than the GF group. The abundances of different gut microbes, including Pseudomonas spp., Orbus spp. and Campylobacter spp., were strongly correlated with amino acid metabolic pathways. Dysgonomonas spp. were strongly correlated with protein digestion and absorption. Issatchenkia spp. had a strong correlation with pepsin activity. Campylobacter spp., Pediococcus spp. and Lactobacillus spp. were strongly correlated with trypsin activity. Lactobacillus spp. and Bacillus spp. were strongly correlated with peptidase activity. Gut microbes such as Issatchenkia spp. may promote the gut proteolytic enzyme activity of BSFL and improve the degradation rate of proteins. BSFL protein digestion and absorption involves gut microbiota that have a variety of functions. In BSFL the core gut microbiota help complete protein degradation. These results demonstrate that core gut microbes in BSFL are important in protein degradation.

An MCIA-like complex is required for mitochondrial complex I assembly and seed development in maize.

During adaptive radiation, mitochondria have co-evolved with their hosts, leading to gain or loss of subunits and assembly factors of respiratory complexes. Plant mitochondrial complex I harbors ∼40 nuclear- and 9 mitochondrial-encoded subunits, and is formed by stepwise assembly during which different intermediates are integrated via various assembly factors. In mammals, the mitochondrial complex I intermediate assembly (MCIA) complex is required for building the membrane arm module. However, plants have lost almost all of the MCIA complex components, giving rise to the hypothesis that plants follow an ancestral pathway to assemble the membrane arm subunits. Here, we characterize a maize crumpled seed mutant, crk1, and reveal by map-based cloning that CRK1 encodes an ortholog of human complex I assembly factor 1, zNDUFAF1, the only evolutionarily conserved MCIA subunit in plants. zNDUFAF1 is localized in the mitochondria and accumulates in two intermediate complexes that contain complex I membrane arm subunits. Disruption of zNDUFAF1 results in severe defects in complex I assembly and activity, a cellular bioenergetic shift to aerobic glycolysis, and mitochondrial vacuolation. Moreover, we found that zNDUFAF1, the putative mitochondrial import inner membrane translocase ZmTIM17-1, and the isovaleryl-coenzyme A dehydrogenase ZmIVD1 interact each other, and could be co-precipitated from the mitochondria and co-migrate in the same assembly intermediates. Knockout of either ZmTIM17-1 or ZmIVD1 could lead to the significantly reduced complex I stability and activity as well as defective seeds. These results suggest that zNDUFAF1, ZmTIM17-1 and ZmIVD1 probably form an MCIA-like complex that is essential for the biogenesis of mitochondrial complex I and seed development in maize. Our findings also imply that plants and mammals recruit MCIA subunits independently for mitochondrial complex I assembly, highlighting the importance of parallel evolution in mitochondria adaptation to their hosts.

Marinobacter daqiaonensis sp. nov., a moderate halophile isolated from a Yellow Sea salt pond.

A Gram-negative, oxidase- and catalase-positive, moderately halophilic strain, designated YCSA40(T), was isolated from sediment of Daqiao saltern in Qingdao, on the east coast of China. Growth occurred at 10-45 °C, at pH 5-9 and with 1-15% NaCl. Strain YCSA40(T) showed the highest 16S rRNA gene sequence similarity to Marinobacter segnicrescens SS011B1-4(T) (97%) and M. gudaonensis SL014B61A(T) (96.9%) and 16S rRNA gene sequence phylogenetic analysis assigned the isolate to the genus Marinobacter. Strain YCSA40(T) contained C(18:1)ω9c (34.8%), C(16:0) (11.6%), C(19:0) cyclo ω10c/C(19:1)ω6c (10.5%), C(16:1)ω9c (8.4%), C(17:0) (6.3%) and C(12:0) 3-OH (5.8%) as the predominant fatty acids. The DNA G+C content was 60.8 mol% and the major ubiquinone was Q-9. These chemotaxonomic characters were all consistent with membership of the genus Marinobacter. DNA-DNA relatedness between the isolate and M. segnicrescens CGMCC 1.6489(T), M. gudaonensis CGMCC 1.6294(T) and other type strains of species of the genus Marinobacter was ≤30%. On the basis of the aforementioned data, it was concluded that strain YCSA40(T) represents a novel species of the genus Marinobacter, for which the name Marinobacter daqiaonensis sp. nov. is proposed. The type strain is YCSA40(T) (=CGMCC 1.9167(T) =NCCB 100308(T) =LMG 25365(T)).

Cohaesibacter marisflavi sp. nov., isolated from sediment of a seawater pond used for sea cucumber culture, and emended description of the genus Cohaesibacter.

A Gram-negative, catalase-negative, oxidase-positive, rod-shaped bacterium, strain DQHS21(T), was isolated from sediment of a seawater pond used for sea cucumber culture at Jimo in Qingdao province on the east coast of China. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain DQHS21(T) belonged to the genus Cohaesibacter, sharing the highest sequence similarity (96.1 %) with Cohaesibacter gelatinilyticus CL-GR15(T), while the similarity to other strains was below 93.0 %. The cellular fatty acids consisted mainly of C(18 : 1)ω7c (60.7 %), C(18 : 0) (17.8 %), C(16 : 0) (8.5 %) and summed feature 3 (C(16 : 1)ω7c and/or iso-C(15 : 0) 2-OH; 6.0 %), which together accounted for 93 % of the total fatty acids. Ubiquinone 10 was the major quinone. The G+C content of the chromosomal DNA of strain DQHS21(T) was 55.2 mol%. The combined genotypic and phenotypic data showed that strain DQHS21(T) represents a novel species of the genus Cohaesibacter, for which the name Cohaesibacter marisflavi sp. nov. is proposed, with the type strain DQHS21(T) ( = CGMCC 1.9157(T)  = NCCB 100300(T)).

Halomonas daqiaonensis sp. nov., a moderately halophilic, denitrifying bacterium isolated from a littoral saltern.

Two novel gram-negative, oxidase- and catalase-positive, rod-shaped bacterial strains, designated YCSA28(T) and YCSA39, were isolated from sediment of Daqiao saltern, Jimo, Qingdao, on the east coast of China. The two strains grew optimally at 28-30 °C, at pH 7.5 and in the presence of 7-8 % (w/v) NaCl. They were assigned to the genus Halomonas, class Gammaproteobacteria, based on 16S rRNA gene sequence analysis. The major cellular fatty acids of the two strains were C(18 : 1)ω7c (42.9 %), C(16 : 0) (23.1 %) and C(16 : 1)ω7c/ω6c (18.0 %), and Q-9 was the major ubiquinone. The G+C content of the DNA of strains YCSA28(T) and YCSA39 was 63.7 and 63.9 mol%, respectively. The predominant respiratory lipoquinone, cellular fatty acid profiles and DNA G+C content of strains YCSA28(T) and YCSA39 were consistent with those of recognized species of the genus Halomonas. Levels of DNA-DNA relatedness between strains YCSA28(T) and YCSA39, between YCSA28(T) and Halomonas ventosae Al12(T), and between YCSA39 and H. ventosae Al12(T) were 95, 45 and 50 %, respectively. Together, these data indicated that strains YCSA28(T) and YCSA39 represent a single novel species of the genus Halomonas, for which the name Halomonas daqiaonensis sp. nov. is proposed. The type strain is YCSA28(T) ( = CGMCC 1.9150(T)  = NCCB 100305(T)  = MCCC 1B00920(T)).

Sunxiuqinia elliptica gen. nov., sp. nov., a member of the phylum Bacteroidetes isolated from sediment in a sea cucumber farm.

Three novel aerobic, elliptic bacteria, designated DQHS4(T), DQHS8 and DQHS15, were isolated from sediment of a seashore pond for sea cucumber culture in Jimo, Qingdao, on the east coast of China. Cells were Gram-, oxidase- and catalase-negative. All three strains grew at 15-42 °C, pH 5-9 and NaCl concentrations between 0.5 and 10%. DNA-DNA hybridization experiments revealed high (>85%) relatedness among the three novel isolates and suggested that the strains constitute a single species. Comparative 16S rRNA gene sequence analysis indicated that these bacteria had less than 90% similarity to all described species of the phylum Bacteroidetes; the closest relative of the three isolates was Prolixibacter bellariivorans F2(T), sharing only 89.6% sequence similarity. The major cellular fatty acids were iso-C(17:0) 3-OH (19.8-20.0%), iso-C(15:0) (16.9-17.3%), anteiso-C(17:1) B and/or iso-C(17:1) I (7.4-8.7%), C(17:0) 2-OH (8.4%), anteiso-C(15:0) (8.2-8.6%) and C(17:1)ω6c (5.6-6.0%). The major respiratory quinone was menaquinone-7 (MK-7) and the DNA G+C content was 41.8-43.5 mol%. Based on the distinct phylogenetic position and the combination of genotypic, phenotypic and chemotaxonomic characteristics, these three strains were considered to represent a novel species of a new genus in the phylum Bacteroidetes, for which the name Sunxiuqinia elliptica gen. nov., sp. nov. is proposed. The type strain of Sunxiuqinia elliptica is DQHS4(T) (=CGMCC 1.9156(T) =NCCB 100301(T) =LMG 25367(T)).

Isolation and Characterization of a Phosphate-Solubilizing Halophilic Bacterium Kushneria sp. YCWA18 from Daqiao Saltern on the Coast of Yellow Sea of China.

Phosphate-solubilizing bacteria (PSB) function in soil phosphorus cycle, increasing the bioavailability of soil phosphorus for plants. Isolation and application of salt-tolerant or halophilic PSB will facilitate the development of saline-alkali soil-based agriculture. A moderately halophilic bacterium was isolated from the sediment of Daqiao saltern on the eastern coast of China, which also performs phosphate-solubilizing ability. The bacterium was assigned to genus Kushneria according to its 16S rRNA gene sequence, and accordingly named as Kushneria sp. YCWA18. The fastest growth was observed when the culturing temperature was 28°C and the concentration of NaCl was 6% (w/v). It was founds that the bacterium can survive at a concentration of NaCl up to 20%. At the optimum condition, the bacterium solubilized 283.16 µg/mL phosphorus in 11 days after being inoculated in 200 mL Ca(3)(PO(4))(2) containing liquid medium, and 47.52 µg/mL phosphorus in 8 days after being inoculated in 200 mL lecithin-containing liquid medium. The growth of the bacterium was concomitant with a significant decrease of acidity of the medium.

IL-23R mutation is associated with ulcerative colitis: A systemic review and meta-analysis.

OBJECTIVES: Since a genome-wide association study revealed that Interleukin-23 receptor (IL-23R) gene is a candidate gene for Ulcerative Colitis (UC), many studies have investigated the association between the IL-23R polymorphisms and UC. However, the results were controversial. The aim of the study was to determine whether the IL-23R polymorphisms confer susceptibility to UC. METHODS: A systematic literature search was carried out to identify all potentially relevant studies. Pooled odds ratios (ORs) with 95% confidence intervals (CIs) were used to estimate the strength of association. RESULTS: A total of 33 studies in 32 articles, including 10,527 UC cases and 15,142 healthy controls, were finally involved in the meta-analysis. Overall, a significant association was found between all UC cases and the rs11209026A allele (OR = 0.665, 95% CI = 0.604~0.733, P < 0.001). Similarly, meta-analyses of the rs7517847, rs1004819, rs10889677, rs2201841, rs11209032, rs1495965, rs1343151 and rs11465804 polymorphisms also indicated significant association with all UC (all P < 0.05). Stratification by ethnicity revealed that the rs11209026, rs7517847, rs10889677, rs2201841 andrs11465804 polymorphisms were associated with UC in the Caucasian group, but not in Asians, while the rs1004819 and rs11209032 polymorphisms were found to be related to UC for both Caucasian and Asian groups. However, subgroup analysis failed to unveil any association between the rs1495965 and rs1343151 polymorphisms and UC in Caucasians or Asians. CONCLUSIONS: The meta-analysis suggests significant association between IL-23R polymorphisms and UC, especially in Caucasians.

Preparation of molecularly imprinted polymers using theanine as dummy template and its application as SPE sorbent for the determination of eighteen amino acids in tobacco.

In this paper, a novel dummy template molecularly imprinted polymer (DMIP) based on a vinyl-SiO2 microspheres surface for the simultaneous selective recognition and enrichment of 18 amino acids was prepared via a surface molecular imprinting technique using theanine as a dummy template. Compared to the imprinted polymers prepared using traditional polymerization techniques, the obtained DMIPs exhibited a regular spherical shape and were relatively monodisperse. The maximal sorption capacity (Qmax) of the resulting DMIPs for the 18 amino acids was up to 1444.3 mg g(-1). A kinetic binding study showed that the sorption capacity reached 85.40% of Qmax in 25 min and sorption equilibrium at 30 min. The imprint factors of the sorbents ranged from 2.86 to 6.9 for the 18 amino acids, which indicated that the DMIP sorbents have high selectivity. An HPLC-UV method for the simultaneous determination of 18 amino acids in tobacco and tobacco smoke was developed using the DMIPs as sorbents for solid phase extraction (SPE) in the sample pretreatment procedure. Under the optimum experimental conditions, the materials had enrichment factors of up to 200 for the amino acids, and the recoveries of the 18 amino acids in tobacco smoke were in the range from 79% to 104% with relative standard deviations of less than 7.4%. It indicated that the obtained DMIP sorbents could specifically recognize the amino acids from complicated samples.