Significance of a minor pilin PilV in biofilm cohesion of Geobacter sulfurreducens.

Bacterial adhesion plays a vital role in forming and shaping the structure of electroactive biofilms that are essential for the performance of bioelectrochemical systems (BESs). Type IV pili are known to mediate cell adhesion in many Gram-negative bacteria, but the mechanism of pili-mediated cell adhesion of Geobacter species on anode surface remains unclear. Herein, a minor pilin PilV2 was found to be essential for cell adhesion ability of Geobacter sulfurreducens since the lack of pilV2 gene depressed the cell adhesion capability by 81.2% in microplate and the anodic biofilm density by 23.1 % at -0.1 V and 37.7 % at -0.3 V in BESs. The less cohesiveness of mutant biofilms increased the charge transfer resistance and biofilm resistance, which correspondingly lowered current generation of the pilV2-deficient strain by up to 63.2 % compared with that of the wild-type strain in BESs. The deletion of pilV2 posed an insignificant effect on the production of extracellular polysaccharides, pili, extracellular cytochromes and electron shuttles that are involved in biofilm formation or extracellular electron transfer (EET) process. This study demonstrated the significance of pilV2 gene in cell adhesion and biofilm formation of G. sulfurreducens, as well as the importance of pili-mediated adhesion for EET of electroactive biofilm.

[Establishment and Evaluation of a Resazurin-Based Microdilution Assay for Microbial Sensitivity Test of Neisseria gonorrhoeae]. / æ·‹ç— å¥ˆç‘ŸèŒè¯æ•è¯•éªŒåˆƒå¤©é’å¾®é‡ç¨€é‡Šæ³•çš„å»ºç«‹ä¸Žè¯„ä»·.

Objective: To establish and evaluate a microbial sensitivity test method for Neisseria gonorrhoeae based on resazurin coloration. Methods: Based on the broth microdilution method, resazurin was added as a live bacteria indicator. WHO G, a WHO gonococcal reference strain, was used to optimize the incubation time for resazurin-stained bacteria and the color change was visually observed to obtain the results. Agar dilution method (the gold standard) and resazurin-based microdilution assay were used to determine the minimum inhibitory concentration (MIC) of azithromycin, ceftriaxone, and spectinomycin for 3 reference strains and 32 isolates of Neisseria gonorrhoeae. The results were analyzed based on essential agreement (EA), which reflected the consistency of the MIC values, category agreement (CA), which reflected the consistency in the determination of drug resistance, intermediary, and sensitivity, very major error (VME), which reflected false sensitivity, and major error (ME), which reflected pseudo drug resistance, to evaluate the accuracy of resazurin-based microdilution assay as a microbial sensitivity test of of Neisseria gonorrhoeae. CA and EA rates≥90% and VME and ME rates≤3% were found to be the acceptable performance rates. Results: The results obtained 6 hours after resazurin was added were consistent with those of the agar dilution method and the resazurin-based microdilution assay was established accordingly based on this parameter. The EA of resazurin-based microdilution assay for measuring the MIC results of azithromycin, ceftriaxone, and spectinomycin was 97.1%, 91.5%, and 94.3%, respectively, and the CA was 88.6%, 94.3%, and 94.3%, respectively. The VME was 0% for all three antibiotics, while the ME was 11.4%, 5.7%, and 5.7%, respectively. Conclusion: The resazurin-based microdilution assay established in this study showed good agreement with agar dilution method for measuring the MIC of antibiotics against Neisseria gonorrhoeae. Moreover, the sensitivity results of this method were highly reliable and could be easily obtained through naked eye observation. Nonetheless, the results of drug resistance should be treated with caution and the optimization of parameters should be continued.

Impact of dietary digestible aromatic amino acid levels and stachyose on growth, nutrient utilization, and cecal odorous compounds in broiler chickens.

This study evaluated the impact of dietary digestible aromatic amino acid (DAAA) levels and stachyose on growth, nutrient utilization and cecal odorous compounds in broiler chickens. A 3×2 two-factor factorial design: Three dietary DAAA levels (1.40, 1.54, 1.68%) supplemented with either 5 g/kg of stachyose or without any stachyose were used to create 6 experimental diets. Each diet was fed to 6 replicates of 10 birds from d 22 to 42. Findings revealed that broilers receiving a diet with 1.54% DAAA levels supplemented with 5 g/kg stachyose exhibited a significant boost in average daily gain and improved utilization of crude protein, ether extract, tryptophan, and methionine compared to other diet treatments (P < 0.05). As the dietary DAAA levels increased, there was a significant rise in the concentrations of indole, skatole, p-methylphenol, and butyric acid in the cecum of broilers (P < 0.05). The addition of stachyose to diets reduced concentrations of indole, skatole, phenol, p-methylphenol, acetic acid and propionic acid in the cecum (P < 0.05). The lowest concentrations of indole, phenol, p-methylphenol, volatile fatty acids and pH in cecum of broilers were observed in the treatment which diet DAAA level was 1.40% with stachyose (P < 0.05). In conclusion, dietary DAAA levels and stachyose had significant interactions on the growth, main nutrient utilization and cecal odorous compounds in broilers. The dietary DAAA level was 1.54% with 5 g/kg of stachyose can improve the growth performance, nutrient utilization. However, the dietary DAAA level was 1.40% with stachyose was more beneficial to decrease the cecal odor compound composition in broilers.

A genetic engineering strategy to enhance outer membrane vesicle-mediated extracellular electron transfer of Geobacter sulfurreducens.

Bioelectrochemical systems (BESs) are unique devices that harness the metabolic activity of electroactive microorganisms (EAMs) to convert chemical energy stored in organic substrates into electrical energy. Enhancing electron transfer efficiency between EAMs and electrodes is the key to practical implementation of BESs. Considering the role of outer membrane vesicles (OMVs) in mediating electron transfer of EAMs, a genetic engineering strategy to achieve OMVs overproduction was explored to enhance electron transfer efficiency and the underlying mechanisms were investigated. This study constructed a mutant strain of Geobacter sulfurreducens that lacked the ompA gene encoding an outer membrane protein. Experimental results showed that the mutant strain produced more OMVs and possessed higher electron transfer efficiency in Fe(III) reduction, dye degradation and current generation in BESs than the wild-type strain. More cargoes such as c-type cytochromes, functional proteins, eDNA, polysaccharides and signaling molecules that might be favorable for electron transfer and biofilm formation were found in OMVs produced by ompA-deficient anodic biofilm, which possibly contributed to the improved electron transfer efficiency of ompA-deficient biofilm. The results indicate that overproduction of OMVs in EAMs might be a potential strategy to enhance BESs performance.

Bidirectional extracellular electron transfer pathways of Geobacter sulfurreducens biofilms: Molecular insights into extracellular polymeric substances.

The basis for bioelectrochemical technology is the capability of electroactive bacteria (EAB) to perform bidirectional extracellular electron transfer (EET) with electrodes, i.e. outward- and inward-EET. Extracellular polymeric substances (EPS) surrounding EAB are the necessary media for EET, but the biochemical and molecular analysis of EPS of Geobacter biofilms on electrode surface is largely lacked. This study constructed Geobacter sulfurreducens-biofilms performing bidirectional EET to explore the bidirectional EET mechanisms through EPS characterization using electrochemical, spectroscopic fingerprinting and proteomic techniques. Results showed that the inward-EET required extracellular redox proteins with lower formal potentials relative to outward-EET. Comparing to the EPS extracted from anodic biofilm (A-EPS), the EPS extracted from cathodic biofilm (C-EPS) exhibited a lower redox activity, mainly due to a decrease of protein/polysaccharide ratio and α-helix content of proteins. Furthermore, less cytochromes and more tyrosine- and tryptophan-protein like substances were detected in C-EPS than in A-EPS, indicating a diminished role of cytochromes and a possible role of other redox proteins in inward-EET. Proteomic analysis identified a variety of redox proteins including cytochrome, iron-sulfur clusters-containing protein, flavoprotein and hydrogenase in EPS, which might serve as an extracellular redox network for bidirectional EET. Those redox proteins that were significantly stimulated in A-EPS and C-EPS might be essential for outward- and inward-EET and warranted further research. This work sheds light on the mechanism of bidirectional EET of G. sulfurreducens biofilms and has implications in improving the performance of bioelectrochemical technology.

Enzymolytic soybean meal improves growth performance, economic efficiency and organ development associated with cecal fermentation and microbiota in broilers offered low crude protein diets.

The objective of this experiment was to determine the effect of low crude protein (CP) diets containing increasing amounts of enzymolytic soybean meal (ESBM) on growth performance, economic benefit and organ development and the role of cecal fermentation and microbiota in broilers. A total of 360 one-day-old Arbor Acres chicks were randomly allocated into 6 groups with 6 replicates and 10 chicks each. The six dietary treatments consisted of a standard high-CP diet (PC), a low-CP diet (NC), and an NC diet with 0.5, 1.0, 1.5%, or 2.0% ESBM. The experiment lasted for 42 days. Compared to PC, NC showed decreased (p < 0.05) average daily gain (ADG) in broilers from 22 to 42 days and from 1 to 42 days, while increasing levels of ESBM quadratically increased (p < 0.05) ADG from 1 to 42 days. Feed cost and total revenue in the NC were lower (p < 0.05) than that in the PC, while supplementation with ESBM in the NC linearly increased (p < 0.05) net profit and economic efficiency in broilers. There were significant differences (p < 0.05) in the liver, proventriculus and gizzard indices between the PC and NC groups, and supplementation with ESBM linearly increased (p < 0.05) the relative weights of liver, pancreas, proventriculus and gizzard in broilers at 42 days of age. The PC group had a higher cecal acetic acid concentration at 21 days and propionic acid concentration at both 21 and 42 days than the NC group (p < 0.05). Cecal acetic acid and propionic acid concentrations linearly increased (p < 0.05) with increasing levels of ESBM in broilers at 42 days of age. No significant differences in ACE, Chao1, Shannon and Simpson indices were observed among groups (p > 0.05), while the cecal abundances of Bacteroides, Faecalibacterium and Clostridium IV increased (p < 0.05) with the increasing level of ESBM in the low-CP diets. In conclusion, feeding ESBM improved economic efficiency, digestive organ development, cecal fermentation and microbial community composition, and up to 2.0% ESBM addition had no negative effect on the growth performance in broilers fed low CP diets.

Characterization of two novel Fe(III)-reducing and electrogenic bacteria, Shewanella ferrihydritica sp. nov. and Shewanella electrica sp. nov., isolated from mangrove sediment.

Three novel strains in the genus Shewanella, designated A3AT, C31T and C32, were isolated from mangrove sediment samples. They were facultative anaerobic, Gram-stain-negative, rod-shaped, flagellum-harbouring, oxidase- and catalase-positive, electrogenic and capable of using Fe(III) as an electron acceptor during anaerobic growth. Results of phylogenetic analysis based on 16S rRNA gene and genomic sequences revealed that the strains should be assigned to the genus Shewanella. The 16S rRNA gene similarity, average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between the isolates and their closely related species were below the respective cut-off values for species differentiation. The 16S rRNA gene similarity, ANI and dDDH values between strains C31T and C32 were 99.7, 99.9 and 99.9 %, respectively, indicating that they should belong to the same genospecies. Based on polyphasic taxonomic approach, two novel species are proposed, Shewanella ferrihydritica sp. nov. with type strain A3AT (GDMCC 1.2732T=JCM 34899T) and Shewanella electrica sp. nov. with type strain C31T (GDMCC 1.2736T=JCM 34902T).

Effects of rumen-protected biotin on the growth performance, nitrogen utilization and blood parameters of yearling Liaoning cashmere doelings.

OBJECTIVE: This study was conducted to investigate the effects of rumen-protected biotin (RPB) on growth performance, nutrient digestibility, nitrogen utilization and plasma biochemical parameters of Liaoning cashmere goats during the cashmere fiber growing period. METHODS: Sixteen 6-month-old Liaoning cashmere twin-doelings (24.8±1.20 kg) were allocated to 2 diet groups that were individually ad libitum fed 30% concentrate and 70% forage diet (dry matter [DM]) by a paired experimental design. Goats of the control group were fed the basal diet, while goats belonging to the RPB group were fed the basal diet with 10 mg RPB/d per animal. The duration of the experiment was 16 weeks with two 8-week periods. Digestibility was determined at weeks 7 and 15, and other measures were taken every four weeks. RESULTS: Compared with the control group, the average daily gain of the RPB group increased by 10.94% (p<0.05), and the intake of neutral detergent fiber was increased (p = 0.045). There were some increasing tendencies for the intake of DM, acid detergent fiber and ether extract (p = 0.070, 0.088, and 0.070, respectively). The intake and digestibility of N tended to increase (p = 0.062 and 0.093, respectively), while the N fecal excretion percentage of N intake was decreased (p = 0.093) in the RPB compared with the control group. N retention tended to increase (p = 0.084) with the addition of adding RPB to the diet. Plasma total protein was increased (p = 0.037), whereas the urea-N concentration was decreased (p = 0.049) in the RPB diet group compared with the control diet group. The levels of propionyl-CoA carboxylase (p<0.001) and methylmalonyl-CoA (p = 0.013) were increased in the RPB group. CONCLUSION: Supplementation of rumen-protected biotin in the diet of cashmere goats can enhance the utilization of N and improve daily weight gain during cashmere fiber growing period.

Effect of enzymolytic soybean meal supplementation on performance, nitrogen excretion, serum biochemical parameters and intestinal morphology in broilers fed low-protein diets.

OBJECTIVE: The objective of this study was to investigate the effect of supplementation with enzymolytic soybean meal (ESBM) on broilers fed low crude protein (CP) diets. METHODS: A total of 360 one-day-old broilers were randomly assigned to six treatments with 6 replicates per treatment and 10 chicks per replicate for a period of 42 days. Chicks were fed a basal standard high-CP diet as a positive control (PC), a low-CP diet (reducing 10 g/kg CP from the PC) as a negative control (NC), or an NC + 0.5%, 1.0%, 1.5%, or 2.0% ESBM diet. RESULTS: Compared to chicks fed the PC, chicks fed the NC had a decreased body weight gain (BWG, p<0.05) from 1 to 42 days, but supplementation with 2.0% ESBM restored BWG (p<0.05) and even linearly improved the feed conversion rate (FCR, p<0.05). Digestibility of CP and ether extract was increased (p<0.05) in chicks fed a 1.0% ESBM diet compared to the PC. With increasing levels of ESBM, nitrogen (N) excretion decreased (p<0.05). The addition of ESBM to the diet did not affect (p>0.05) serum concentrations of total protein, albumin and total cholesterol but led to a descending trend in triglycerides and an ascending trend in calcium and urea N at 42 days (p<0.10). There were no differences (p>0.05) in villus height (VH), crypt depth (CD), and VH/CD (V/C) of the duodenum and jejunum between the PC and NC at both 21 days and 42 days, while increasing dietary ESBM levels linearly (p<0.05) decreased CD and increased V/C of the duodenum and jejunum at both 21 days and 42 days. CONCLUSION: The findings indicated that ESBM could be used in broiler low-CP diets to improve production performance, decrease N excretion, and enhance intestinal health.

Microbiome-metabolomics analysis reveals abatement effects of itaconic acid on odorous compound production in Arbor Acre broilers.

BACKGROUND: Public complaints concerning odor emissions from intensive livestock and poultry farms continue to grow, as nauseous odorous compounds have adverse impacts on the environment and human health. Itaconic acid is a metabolite from the citric acid cycle of the host and shows volatile odor-reducing effects during animal production operations. However, the specific role of itaconic acid in decreasing intestinal odorous compound production remains unclear. A total of 360 one-day-old chicks were randomly divided into 6 treatment groups: control group (basal diet) and itaconic acid groups (basal diet + 2, 4, 6, 8 and 10 g/kg itaconic acid). The feeding experiment lasted for 42 d. RESULTS: Dietary itaconic acid supplementation linearly and quadratically decreased (P < 0.05) the cecal concentrations of indole and skatole but did not affect (P > 0.05) those of lactic, acetic, propionic and butyric acids. The cecal microbial shift was significant in response to 6 g/kg itaconic acid supplementation, in that the abundances of Firmicutes, Ruminococcus and Clostridium were increased (P < 0.05), while those of Bacteroidetes, Escherichia-Shigella and Bacteroides were decreased (P < 0.05), indicative of increased microbial richness and diversity. Furthermore, a total of 35 significantly (P < 0.05) modified metabolites were obtained by metabolomic analysis. Itaconic acid decreased (P < 0.05) the levels of nicotinic acid, nicotinamide, glucose-6-phosphate, fumatic acid and malic acid and increased (P < 0.05) 5-methoxytroptomine, dodecanoic acid and stearic acid, which are connected with the glycolytic pathway, citrate acid cycle and tryptophan metabolism. Correlation analysis indicated significant correlations between the altered cecal microbiota and metabolites; Firmicutes, Ruminococcus and Clostridium were shown to be negatively correlated with indole and skatole production, while Bacteroidetes, Escherichia-Shigella and Bacteroides were positively correlated with indole and skatole production. CONCLUSIONS: Itaconic acid decreased cecal indole and skatole levels and altered the microbiome and metabolome in favor of odorous compound reduction. These findings provide new insight into the role of itaconic acid and expand its application potential in broilers.

Sub-MIC antibiotics affect microbial ferrihydrite reduction by extracellular membrane vesicles.

Environmental concentrations of antibiotics, usually below MIC, have significant biological effects on bacterial cells. Sub-MIC antibiotics exposure induces bacteria to produce outer membrane vesicles (OMVs). Recently, OMVs is discovered as a novel pathway for dissimilatory iron reducing bacteria (DIRB) to mediate extracellular electron transfer (EET). Whether and how the antibiotic-induced OMVs modulate iron oxides reduction by DIRB have not been studied. This study showed the sub-MIC antibiotics (ampicillin or ciprofloxacin) increased OMVs secretion in Geobacter sulfurreducens, and the antibiotic-induced OMVs contained more redox active cytochromes facilitating iron oxides reduction, especially for the ciprofloxacin-induced OMVs. Deduced from a combination of electron microscopy and proteomic analysis, the influence of ciprofloxacin on SOS response triggered prophage induction and led to the formation of outer-inner membrane vesicles (OIMVs) in, which was a first report in Geobacter species. While ampicillin disrupting cell membrane integrity resulted in more formation of classic OMVs from outer membrane blebbing. The results indicated that the different structure and composition of vesicles were responsible for the antibiotic-dependent regulation on iron oxides reduction. This newly identified regulation on EET-mediated redox reactions by sub-MIC antibiotics expands our knowledge about the impact of antibiotics on microbial processes or "non-target" organisms.

Effects of fermented feed on growth performance, immune organ indices, serum biochemical parameters, cecal odorous compound production, and the microbiota community in broilers.

The aim of this study was to explore the effects of dietary fermented feed addition on growth performance, immune organ indices, serum biochemical parameters, cecal odorous compound production, and the bacterial community in broilers. A total of 480 broiler chicks (1-day-old) were randomly assigned to 6 groups, including a basal diet (control group), a basal diet supplemented with 10, 15, 20, and 25% dried fermented feed, and 10% wet fermented feed. Each group contained 8 replicates of 10 chicks each. The results showed that fermentation increased (P < 0.05) the total acid level and the number of Lactobacillus, Yeast, and Bacillus. The 15% dried fermented feed group had an increased (P < 0.05) body weight (BW) than the control, while the 25% dried fermented feed group had the lowest (P < 0.05) BW on 42 d. Compared to the control group, the feed intake (FI) was increased (P < 0.05) in the 10, 15% dried and 10% wet fermented feed groups from 22 to 42 d and from 1 to 42 d. No significant difference (P > 0.05) was observed in feed conversion ratio (FCR) among all groups. Supplementation with fermented feed increased (P < 0.05) the bursa of Fabricius index but not (P > 0.05) the thymus and spleen indices. Compared with the control, the broilers fed fermented feed had increased (P < 0.05) serum total protein, albumin, globulin, IgA, IgG, IgM, lysozyme, complement 3, and complement 4 levels. The cecal concentrations of acetic acid, propionic acid, butyric acid, and lactic acid were increased and the pH values were decreased in the fermented feed groups (P < 0.05). Among the groups, the 15% dried fermented feed group showed the lowest concentrations of skatole and indole in the cecum (P < 0.05). The composition of the cecal microbiota was characterized, in which an increased abundance of Ruminococcaceae, Lactobacillaceae, and unclassified Clostridiales and a decreased abundance of Rikenellaceae, Lachnospiraceae, and Bacteroidaceae were found in the fermented feed groups. Taken together, dietary fermented feed supplementation can improve growth performance, immune organ development, and capacity and decrease cecal odorous compound production, which may be related to the regulation of microbial composition.

Sub-inhibitory concentrations of ampicillin affect microbial Fe(III) oxide reduction.

Antibiotics are ubiquitous in the iron-rich environments but their roles in microbial reduction of Fe(III) oxides are still unclear. Using ampicillin and Geobacter soli, this study investigated the underlying mechanism by which antibiotic regulated microbial reduction of Fe(III) oxides. Results showed that sub-minimal inhibitory concentrations (sub-MIC) of ampicillin significantly affected ferrihydrite reduction by G. soli, with a stimulatory effect at 1/64 and 1/32 MIC and an inhibitory effect at 1/8 MIC. Increasing ampicillin concentration resulted in increasing cell length and decreasing bacterial zeta potential that were beneficial for ferrihydrite reduction, and decreasing outer membrane permeability that was unfavorable for ferrihydrite reduction. The respiratory metabolism ability was enhanced by 1/64 and 1/32 MIC ampicillin and reduced by 1/8 MIC ampicillin, which was also responsible for regulation of ferrihydrite reduction by ampicillin. The ferrihydrite reduction showed a positive correlation with the redox activity of extracellular polymeric substances (EPS) which was tied to the cytochrome/polysaccharide ratio and the content of α-helices and ß-sheet in EPS. These results suggested that ampicillin regulated microbial Fe(III) oxide reduction through modulating the bacterial morphology, metabolism activity and extracellular electron transfer ability. Our findings provide new insights into the environmental factors regulating biogeochemical cycling of iron.

Interaction between dietary digestible tryptophan and soy oligosaccharides in broiler chickens: effects on caecal skatole level and microflora.

OBJECTIVE: This study was conducted to evaluate the interactive effects of dietary digestible tryptophan (dTry) and soy oligosaccharides (SO) on growth performance, caecal skatole level, and microflora of broiler chickens aged from 14 to 42 days. METHODS: Three hundred and sixty broiler chicks were allocated equally to 36 cages at 14-dayof- age according to body weight and gender. Using a 3×2 factorial arrangement, 3 dietary dTry levels (0.18%, 0.23%, and 0.28%) supplemented with 0 or 3.5 g/kg of SO were used to create 6 diets (treatments). Each diet was fed to six replicates of 10 birds (60 birds/treatment), growth performance was measured. Caecal content samples were collected at 42 days of age. RESULTS: Results showed that significantly different dTry level×SO interactions were found for average daily gain (ADG), caecal levels of indole, propionic acid, and butyric acid, and microbial Shannon index (p<0.05). Birds fed diet containing 0.23% dTry level with SO supplementation had higher ADG and lower feed/gain ratio than those fed the other diets (p<0.05). Broilers fed diets containing 0.28% dTry increased their caecal levels of indole and skatole compared with those containing 0.18% or 0.23% dTry (p<0.01), regardless of SO addition. SO supplementation to diets decreased the caecal skatole level by 16.17% (p<0.05), and increased the relative frequency of Clostridium IV (p<0.05), regardless of dietary dTry level. CONCLUSION: These results indicated that diets containing 0.23% dTry with SO supplementation positively promoted ADG, and decreased caecal skatole levels of broiler chickens. The dietary dTry level, SO affected the caecal skatole level, however, there was no interaction between them.

Shewanella shenzhenensis sp. nov., a novel Fe(III)-reducing bacterium with abundant possible cytochrome genes, isolated from mangrove sediment.

A facultative anaerobic bacterium, designated as A25T, was isolated from a mangrove sediment sample collected in Shenzhen, China. Cells of strain A25T were found to be Gram-staining negative, rod-shaped, flagella-harboring, and oxidase- and catalase-positive. The isolate was able to grow at 4-40 °C (optimum 28 °C) and pH 5.0-9.0 (optimum pH 6.0), and in 0-10% NaCl concentration (w/v) (optimum 1%). Strain A25T was capable of reducing Fe(III) citrate under anaerobic conditions. The major fatty acids of this strain was C16:1ω7c/C16:1ω6c (summed feature 3), C17:1ω8c and iso-C15:0. Results of phylogenetic analyses based on 16S rRNA gene sequences indicated that strain A25T is affiliated with the genus Shewanella, showing the highest similarity to Shewanella seohaensis S7-3T (98.4% similarity). The average nucleotide identity and digital DNA-DNA hybridization values between the genomes of strain A25T and its closely related strains were ≤ 79.0% and ≤ 22.8%, respectively. Based on its phenotypic, phylogenetic properties and physiological and biochemical characteristics, strain A25T (= JCM 34900T = GDMCC 1.2731T) was designated as the type strain of a novel species of the genus Shewanella, for which the name Shewanella shenzhenensis sp. nov. was proposed.

[Contamination Characteristics and Ecological Risk Assessment of Pharmaceuticals and Personal Care Products (PPCPs) in the Third Drain of Ningxia].

To investigate the contamination characteristics and potential ecological risks of pharmaceuticals and personal care products (PPCPs) in the Third Drain of Ningxia, 14 PPCPs were detected and analyzed using solid-phase extraction coupled with ultra-high performance liquid chromatography-tandem mass spectrometry. The results showed that these 14 PPCPs were detected in the Third Drain and its confluent streams, with total concentrations of 117.74-1947.64 ng·L-1 and 63.94-4509.39 ng·L-1, respectively. Detection ratios of gemfibrozil (GEM), caffeine (CAF), avobenzone (BM-DBM), benzophenone-3 (BP-3), 3-(4-methylbenzylidene)-camphor (4-MBC), and diethyltoluamide (DEET) were 100% in the drain. The highest-concentration pharmaceutical was GEM (7.78-721.84 ng·L-1), followed by CAF (41.74-246.86 ng·L-1), and the highest-concentration personal care product was DEET (3.17-219.91 ng·L-1), followed by BP-3 (56.92-150.14 ng·L-1). Concentrations of PPCPs at different sampling points exhibited spatial differences. The total PPCPs concentration increased dramatically and reached a maximum value after flowing through Pingluo County, then showed a decreasing trend downstream. Correlation analysis showed that 4-MBC was significantly positively correlated with COD (P<0.01). IBU, XMTD, TCC, and TCS were significantly correlated with NH4+-N (P<0.05). DIC, BF, CBZ, and DEET were significantly correlated with TN (P<0.05). The results indicated that concentrations of PPCPs were closely related to water quality indexes. Risk assessment showed that DIC, IBU, GEM, CBZ, CAF, and BP-3 had high risks, whereas BM-DBM, TCC, and TCS had moderate risks.

The Role of Exopolysaccharides in Direct Interspecies Electron Transfer.

Direct interspecies electron transfer (DIET) is an effective mechanism for microbial species to exchange electrons cooperatively during syntrophic metabolism. It is generally accepted that DIET is mainly mediated by electrically conductive pili and outer surface c-type cytochromes (c-Cyts). However, as an extracellular matrix is ubiquitous and abundant on the surface of microorganisms, the effect and mechanism of exopolysaccharides on DIET are still unclear. This study constructed a co-culture of exopolysaccharides-deficient Geobacter sulfurreducens with Geobacter metallireducens to explore the role of exopolysaccharides in DIET. Results revealed that the deficiency of exopolysaccharides extended the metabolic period of the co-culture by 44.4% and changed the proportions of each species in the co-culture. The exopolysaccharides-deficient co-culture failed to form large, tight spherical aggregates and the expression of c-Cyts and pili was decreased. The addition of magnetite and granular activated carbon (GAC), respectively, might compensate for the functions of c-Cyts and pili in the first generation of co-culture, but the stimulatory effect on the metabolic stable period co-culture was fairly limited. These findings demonstrate that non-conductive exopolysaccharides are an important component of DIET aggregates and an extracellular matrix for DIET-required c-Cyts.

Geobacter benzoatilyticus sp. nov., a novel benzoate-oxidizing, iron-reducing bacterium isolated from petroleum contaminated soil.

A strictly anaerobic bacterial strain, designated Jerry-YXT, was isolated from petroleum-contaminated soil sampled in China. Strain Jerry-YXT was a Gram-stain-negative bacterium forming reddish colonies. It grew optimally at 30 °C and pH 7.0, and tolerated 1.0 % (w/v) NaCl. Strain Jerry-YXT was able to use fumarate, ferric citrate and ferrihydrite as electron acceptors, and ethanol, acetate and benzoate as electron donors. The major fatty acids of this strain were C16 : 0 and C16 : 1 ω7c/C16 : 1 ω6c (summed feature 3). The 16S rRNA gene sequence-based phylogenetic analysis placed this strain in the genus Geobacter, being most closely related to Geobacter metallireducens (98.2 % similarity), Geobacter hydrogenophilus (98.1 %) and Geobacter grbiciae (98.0 %). The DNA G+C content was 57.6 mol%. The average nucleotide identity and digital DNA-DNA hybridization values between the genomes of strain Jerry-YXT and G. metallireducens GS-15T were 81.8 and 35.4 %, respectively. The results of the polyphasic study allowed the genotypic and phenotypic differentiation of strain Jerry-YXT from its closest species, which suggested that strain Jerry-YXT represents a novel species of the genus Geobacter. The name for the proposed new species is Geobacter benzoatilyticus sp. nov. The type strain is Jerry-YXT (=MCCC 1K05659T=JCM 39190T).

[Contamination Characteristics and Ecological Risk Assessment of Antibiotics in the Third Drain of Ningxia].

Solid-phase extraction and ultra-high performance liquid chromatography-electrostatic field orbit hydrazine mass spectrometry were used to detect and analyze the contamination of nine antibiotics including sulfonamides, tetracyclines, macrolides, quinolones, and lincomycin in the third drain of Ningxia. Correlations between the antibiotic concentration and water quality indexes were explored, and the ecological risks were evaluated. The results showed that seven antibiotics were detected in the third drain and its confluent stream, with the total concentrations of 14.91-153.48 ng·L-1 and ND-39.37 ng·L-1, respectively. The detection ratios of clarithromycin, levofloxacin, ciprofloxacin, and lincomycin were 100% in the third drain. The highest-concentration antibiotic was levofloxacin (0.84-94.12 ng·L-1), followed by lincomycin (11.15-48.13 ng·L-1). Based on the spatial distribution analysis, after flowing through Pingluo County, total antibiotic concentrations in the third drain increased significantly and showed an increasing trend. The maximum concentration appeared in the Huinong section and showed a decreasing trend downstream. The total concentration of antibiotics before flowing into the Yellow River was 20.26 ng·L-1. Correlation analysis showed that levofloxacin was significantly positively correlated with NH4+-N and TN (P<0.01), erythromycin was significantly correlated with TN, and clarithromycin was significantly correlated with both NH4+-N and TN (P<0.05), which indicated that water quality index was closely related to antibiotic concentration. The results of the ecological risk assessment showed that levofloxacin and clarithromycin in the third drain posed certain ecological risks.

Effects of dietary supplementation with itaconic acid on the growth performance, nutrient digestibility, slaughter variables, blood biochemical parameters, and intestinal morphology of broiler chickens.

Itaconic acid (IA) is a biologically based unsaturated dicarboxylic acid secreted by mammalian cells. While IA has potential for use in multiple applications, information regarding the influence of IA on animal production remains scarce. This study investigated the effects of dietary IA supplementation on the growth performance, nutrient digestibility, slaughter variables, blood parameters, and intestinal morphology of broiler chickens. A total of 360 one-day-old Arbor Acre broiler chicks were allotted to 6 groups, with 10 chicks per cage and 6 replicates per group in a randomized complete block design. Broiler chicks were fed a basal diet with 0 (control), 0.2, 0.4, 0.6, 0.8, or 1.0% IA. The experimental period lasted from 1 to 42 d of age. Dietary IA supplementation did not affect average daily gain (ADG) and feed/gain ratio (F/G) but quadratically increased average daily feed intake (ADFI) and linearly increased crude protein (CP) digestibility during the grower period (d 22-42). A higher breast and thigh muscle yield and a lower abdominal fat yield were observed in a linear and quadratic manner with the IA supplementation. Adding IA to the diet had significant effects on superoxide dismutase (SOD), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and catalase (CAT) levels in serum at d 21 and on total antioxidation capacity (T-AOC) at d 42. There were linear and quadratic increases in villus height and the villus height/crypt depth ratio (V/C) of the duodenum and villus height of the jejunum with the supplementation of IA. Regression analyses for ADFI, dressed yield, breast and thigh muscle yield, abdominal fat yield, serum ALT, CAT, and SOD levels, villus length of the duodenum and jejunum, and V/C of the duodenum indicated that the optimal dietary IA supplementation would be from 0.4 to 0.7%. From an economic perspective, a level of 0.4% IA in the broiler diet is recommended for improving the nutrient digestibility, slaughter performance, antioxidant ability, and intestinal morphology of broiler chickens.