Identification of Bacterial Fish Pathogens in Brazil by Direct Colony PCR and 16S rRNA Gene Sequencing

Intensive fish farming systems in Brazil have increased the disease incidence, mainly of bacterial origin, due to higher stocking density, high organic matter levels and poor quality of the aquatic environment that causes high mortality rates during outbreaks. The identification of pathogenic species using a fast and reliable method of diagnosis is essential for successful epidemiological studies and disease control. The present study evaluated the use of direct colony PCR in combination with 16S rRNA gene sequencing to diagnose fish bacterial diseases, with the goal of reducing the costs and time necessary for bacterial identification. The method was successful for all 178 isolates tested and produced bands with the same intensity as the standard PCR performed using pure DNA. In conclusion, the genetics methods allowed detecting the most common and important pathogens in Aquaculture, including 12 species of occurrence in Brazilian fish farms. The results of the present study constitute an advance in the available diagnostic methods for bacterial pathogens in fish farms.

Enterococcus faecalis provides protection during scavenging in carrion crow (Corvus corone)

The gut microbiota significantly influences host physiology and provides essential ecosystem services.While diet can affect the composition of the gut microbiota,the gut microbiota can also help the host adapt to specific dietary habits.The carrion crow(Corvus corone),an urban facultative scavenger bird,hosts an abundance of pathogens due to its scavenging behavior.Despite this,carrion crows infrequently exhibit illness,a phenomenon related to their unique physiological adaptability.At present,however,the role of the gut microbiota remains incompletely understood.In this study,we performed a comparative analysis using 16S rRNA amplicon sequencing technology to assess colonic content in carrion crows and 16 other bird species with different diets in Beijing,China.Our findings revealed that the dominant gut microbiota in carrion crows was primarily composed of Proteobacteria(75.51%)and Firmicutes(22.37%).Significant differences were observed in the relative abundance of Enterococcus faecalis among groups,highlighting its potential as a biomarker of facultative scavenging behavior in carrion crows.Subsequently,E.faecalis isolated from carrion crows was transplanted into model mice to explore the protective effects of this bacterial community against Salmonella enterica infection.Results showed that E.faecalis down-regulated the expression of pro-inflammatory cytokines tumor necrosis factor alpha(TNF-α),interferon gamma(IFN-γ),and interleukin 6(IL-6),prevented S.enterica colonization,and regulated the composition of gut microbiota in mice,thereby modulating the host’s immune regulatory capacity.Therefore,E.faecalis exerts immunoregulatory and anti-pathogenic functions in carrion crows engaged in scavenging behavior,offering a representative case of how the gut microbiota contributes to the protection of hosts with specialized diets.

Alterations in gut microbiota are related to metabolite profiles in spinal cord injury

Studies have shown that gut microbiota metabolites can enter the central nervous system via the blood-spinal cord barrier and cause neuroinflammation, thus constituting secondary injury after spinal cord injury. To investigate the correlation between gut microbiota and metabolites and the possible mechanism underlying the effects of gut microbiota on secondary injury after spinal cord injury, in this study, we established mouse models of T8–T10 traumatic spinal cord injury. We used 16 S rRNA gene amplicon sequencing and metabolomics to reveal the changes in gut microbiota and metabolites in fecal samples from the mouse model. Results showed a severe gut microbiota disturbance after spinal cord injury, which included marked increases in pro-inflammatory bacteria, such as Shigella, Bacteroides, Rikenella, Staphylococcus, and Mucispirillum and decreases in anti-inflammatory bacteria, such as Lactobacillus, Allobaculum, and Sutterella. Meanwhile, we identified 27 metabolites that decreased and 320 metabolites that increased in the injured spinal cord. Combined with pathway enrichment analysis, five markedly differential amino acids(L-leucine, L-methionine, L-phenylalanine, L-isoleucine and L-valine) were screened out, which play a pivotal role in activating oxidative stress and inflammatory responses following spinal cord injury. Integrated correlation analysis indicated that the alteration of gut microbiota was related to the differences in amino acids, which suggests that disturbances in gut microbiota might participate in the secondary injury through the accumulation of partial metabolites that activate oxidative stress and inflammatory responses. Findings from this study provide a new theoretical basis for improving the secondary injury after spinal cord injury through fecal microbial transplantation.

Alterations in the human oral microbiome in cholangiocarcinoma

Dear Editor,Alterations in the human microbiome are closely related to various hepatobiliary diseases.Gut microbial dysbiosis has been found in patients with cholangiocarcinoma(CCA)[1].However,the characteristics of oral microbiome in patients with CCA have not been studied.

Sarcopenia and gut microbiota alterations in patients with hematological diseases before and after hematopoietic stem cell transplantation

Objective: The aim of this study was to investigate the prevalence of sarcopenia(SP) and its relationship with gut microbiota alterations in patients with hematological diseases before and after hematopoietic stem cell transplantation(HSCT).Methods: A total of 108 patients with various hematological disorders were selected from Peking University People’s Hospital. SP was screened and diagnosed based on the 2019 Asian Sarcopenia Diagnosis Strategy. Physical measurements and fecal samples were collected, and 16S rRNA gene sequencing was conducted. Alpha and beta diversity analyses were performed to evaluate gut microbiota composition and diversity.Results: After HSCT, significant decreases in calf circumference and body mass index(BMI) were observed,accompanied by a decline in physical function. Gut microbiota analyses revealed significant differences in the relative abundance of Enterococcus, Bacteroides, Blautia and Dorea species before and after HSCT(P<0.05). Before HSCT, sarcopenic patients had lower Dorea levels and higher Phascolarctobacterium levels than non-sarcopenia patients(P<0.01). After HSCT, no significant differences in species abundance were observed. Alpha diversity analysis showed significant differences in species diversity among the groups, with the highest diversity in the postHSCT 90-day group and the lowest in the post-HSCT 30-day group. Beta diversity analysis revealed significant differences in species composition between pre-and post-HSCT time points but not between SP groups. Linear discriminant analysis effect size(LEfSe) identified Alistipes, Rikenellaceae, Alistipes putredinis, Prevotellaceae defectiva and Blautia coccoides as biomarkers for the pre-HSCT sarcopenia group. Functional predictions showed significant differences in anaerobic, biofilm-forming and oxidative stress-tolerant functions among the groups(P<0.05).Conclusions: This study demonstrated a significant decline in physical function after HSCT and identified potential gut microbiota biomarkers and functional alterations associated with SP in patients with hematological disorders. Further research is needed to explore the underlying mechanisms and potential therapeutic targets.

Effects of reclaimed water irrigation and nitrogen fertilization on the chemical properties and microbial community of soil

The ecological effect of reclaimed water irrigation and fertilizer application on the soil environment is receiving more attention.Soil microbial activity and nitrogen（N）levels are important indicators of the effect of reclaimed water irrigation on environment.This study evaluated soil physicochemical properties and microbial community structure in soils irrigated with reclaimed water and receiving varied amounts of N fertilizer.The results indicated that the reclaimed water irrigation increased soil electrical conductivity（EC）and soil water content（SWC）.The N treatment has highly significant effect on the ACE,Chao,Shannon（H）and Coverage indices.Based on a 16S ribosomal RNA（16S rRNA）sequence analysis,the Proteobacteria,Gemmatimonadetes and Bacteroidetes were more abundant in soil irrigated with reclaimed water than in soil irrigated with clean water.Stronger clustering of microbial communities using either clean or reclaimed water for irrigation indicated that the type of irrigation water may have a greater influence on the structure of soil microbial community than N fertilizer treatment.Based on a canonical correspondence analysis（CCA）between the species of soil microbes and the chemical properties of the soil,which indicated that nitrate N（NO3-–-N）and total phosphorus（TP）had significant impact on abundance of Verrucomicrobia and Gemmatimonadetes,meanwhile the p H and organic matter（OM）had impact on abundance of Firmicutes and Actinobacteria significantly.It was beneficial to the improvement of soil bacterial activity and fertility under 120 mg kg^-1 N with reclaimed water irrigation.

Effects of sepsis and its treatment measures on intestinal flora structure in critical care patients

BACKGROUND Sepsis is a common disease in intensive care units,with high morbidity and mortality.Intestinal microecology plays a vital part in the development and progression of this disease,possibly because sepsis and its treatment cause specific changes in the composition of the intestinal flora.AIM To investigate the characteristics of intestinal flora disturbance in sepsis patients treated with antibiotics.METHODS In this prospective comparative study,we enrolled ten patients with sepsis(sepsis group),hospitalized in the Department of Critical Care Medicine of the General Hospital,Ningxia Medical University,China(a class IIIa general hospital)from February 2017 to June 2017;ten patients without sepsis hospitalized in the same period(non-sepsis group)and ten healthy individuals(control group)were also enrolled.Fecal samples collected from the three groups were subjected to 16S rRNA gene sequencing and the intestinal flora diversity,structure,and composition were determined.Additionally,the dynamics of the intestinal flora diversity,structure,and composition in sepsis patients were investigated via 16S rRNA gene sequencing of samples collected 0 d,3 d,and 7 d after admittance to the intensive care unit.Correlations between the serum levels of procalcitonin,endotoxin,diamine oxidase,and D-lactic acid and the intestinal flora composition of sepsis patients were also investigated.RESULTS Compared with the healthy control group,sepsis and non-sepsis patients showed reduced intestinal floraα-diversity and a distinct flora structure,with Firmicutes as the dominant phylum,and significantly decreased proportions of Bacteroidetes,as well as Prevotella and Lachnospira,among other genera.Of note,the proportion of Enterococcus was significantly increased in the intestinal tract of sepsis patients.Interestingly,theα-diversity in the sepsis group decreased gradually,from days 1 to 7 of treatment.However,pairwise comparisons showed that both the diversity and structure of the intestinal flora were not significantly different considering the three different time points studied.Curiously,the serum levels of procalcitonin,endotoxin,diamine oxidase,and D-lactic acid in sepsis patients correlated with the prevalence of various bacterial genera.For example,the prevalence of Ruminococcus was positively correlated with serum procalcitonin,endotoxins,and diamine oxidase;similarly,the prevalence of Roseburia was positively correlated with serum procalcitonin,endotoxins,and D-lactic acid.CONCLUSION Sepsis patients in intensive care units show dysbiosis,lasting for at least 1 wk.

Relationship between hyperlipidemia and the gut microbiome of rats, characterized using high-throughput sequencing

Objective:To determine the effects of a high-fat diet(HFD)on the gut microbiome in rats,to explore the relationship between the intestinal flora and blood lipid profile.Methods:SpragueeDawley rats were fed an HFD for four weeks to induce hyperlipidemia,then 16S rRNA sequencing was used to compare the intestinal flora between hyperlipidemic and control diet-fed rats.Results:The microbiome of rats fed an HFD for four weeks differed from that of control diet-fed rats.Bacterial species that were less abundant were most affected by HFD feeding,among which were many pathogenic species,which became significantly more abundant.Eighteen genera were present in significantly different numbers in hyperlipidemic and control rats,more than half of which have been linked to infection and inflammation,or energy intake and obesity.The results indicated a type of stress response of the flora to a high-fat environment.In addition,the age of the rats tended to influence the gut microbial composition.Conclusion:These findings suggest that HFD may induce hyperlipidemia by affecting the gut microbial composition.Changes in the abundance of pro-inflammatory and pathogenic bacteria,and those that influence energy intake and obesity,may be important mediators of this.

A novel apidaecin Api-PR19 synergizes with the gut microbial community to maintain intestinal health and promote growth performance of broilers

Background: Antibiotic growth promoters(AGPs) have been used as growth promoters to maintain animal intestinal health and improve feed efficiency in broilers by inhibiting pathogen proliferation. In view of the growing emergence of antibiotic-resistant pathogen strains and drug residue issues, novel treatments are increasingly required. This study aimed to compare two antimicrobial approaches for managing pathogen infection and maintaining animal intestinal health in broilers by supplying Apidaecin Api-PR19 and AGPs over 42 d of a feeding trial.Results: Compared with the broilers that were only fed a corn-soybean basal diet(CON group), supplementation with Api-PR19 and AGP(respectively named the ABP and AGP groups) both increased the feed conversion efficiency. When compared with the AGP group, Api-PR19 supplementation could significantly increase the organ index of the bursa of fabricius and subtype H9 antibody level in broiler chickens. Moreover, when compared with the CON group, the intestinal villus height, intestinal nutrient transport, and intestinal s Ig A content were all increased in the Api-PR19 group, while AGP supplementation was harmful to the intestinal villus height and intestinal nutrient transport. By assessing the antibacterial effect of Api-PR19 and antibiotics in vitro and in vivo, we found that Api-PR19 and antibiotics both inhibited the growth of pathogens, including Escherichia coli and Campylobacter jejuni. Furthermore, by using 16 S r RNA gene sequencing, the beneficial bacteria and microbiota in broilers were not disturbed but improved by apidaecin Api-PR19, including the genera of Eubacterium and Christensenella and the species of uncultured_Eubacterium_sp, Clostridium_asparagiforme, and uncultured_Christensenella_sp, which were positively related to improved intestinal development, absorption, and immune function.Conclusion: Apidaecin Api-PR19 treatment could combat pathogen infection and had little negative impact on beneficial bacteria in the gut compared to antibiotic treatment, subsequently improving intestinal development,absorption, and immune function.

Gut mucosal microbiota profiles linked to colorectal cancer recurrence

BACKGROUND Emerging evidence links gut microbiota to various human diseases including colorectal cancer(CRC)initiation and development.However,gut microbiota profiles associated with CRC recurrence and patient prognosis are not completely understood yet,especially in a Chinese cohort.AIM To investigate the relationship between gut mucosal microbiota profiles and CRC recurrence and patient prognosis.METHODS We obtained the composition and structure of gut microbiota collected from 75 patients diagnosed with CRC and 26 healthy controls.The patients were followed up by regular examination to determine whether tumors recurred.Triplet-paired samples from on-tumor,adjacent-tumor and off-tumor sites of patients diagnosed with/without CRC recurrence were analyzed to assess spatial-specific patterns of gut mucosal microbiota by 16S ribosomal RNA sequencing.Next,we carried out bioinformatic analyses,Kaplan-Meier survival analyses and Cox regression analyses to determine the relationship between gut mucosal microbiota profiles and CRC recurrence and patient prognosis.RESULTS We observed spatial-specific patterns of gut mucosal microbiota profiles linked to CRC recurrence and patient prognosis.A total of 17 bacterial genera/families were identified as potential biomarkers for CRC recurrence and patient prognosis,including Anaerotruncus,Bacteroidales,Coriobacteriaceae,Dialister,Eubacterium,Fusobacterium,Filifactor,Gemella,Haemophilus,Mogibacteriazeae,Pyramidobacter,Parvimonas,Porphyromonadaceae,Slackia,Schwartzia,TG5 and Treponema.CONCLUSION Our work suggests that intestinal microbiota can serve as biomarkers to predict the risk of CRC recurrence and patient death.

Salmonella Typhimurium infection disrupts but continuous feeding of Bacillus based probiotic restores gut microbiota in infected hens

Background:The gut microbiota plays an important role in the colonisation resistance and invasion of pathogens.Salmonella Typhimurium has the potential to establish a niche by displacing the microbiota in the chicken gut causing continuous faecal shedding that can result in contaminated eggs or egg products.In the current study,we investigated the dynamics of gut microbiota in laying chickens during Salmonella Typhimurium infection.The optimisation of the use of an infeed probiotic supplement for restoration of gut microbial balance and reduction of Salmonella Typhimurium load was also investigated.Results:Salmonella infection caused dysbiosis by decreasing(FDR<0.05)the abundance of microbial genera,such as Blautia,Enorma,Faecalibacterium,Shuttleworthia,Sellimonas,Intestinimonas and Subdoligranulum and increasing the abundance of genera such as Butyricicoccus,Erysipelatoclostridium,Oscillibacter and Flavonifractor.The higher Salmonella Typhimurium load resulted in lower(P<0.05)abundance of genera such as Lactobacillus,Alistipes,Bifidobacterium,Butyricimonas,Faecalibacterium and Romboutsia suggesting Salmonella driven gut microbiota dysbiosis.Higher Salmonella load led to increased abundance of genera such as Caproiciproducens,Acetanaerobacterium,Akkermansia,Erysipelatoclostridium,Eisenbergiella,EscherichiaShigella and Flavonifractor suggesting a positive interaction of these genera with Salmonella in the displaced gut microbiota.Probiotic supplementation improved the gut microbiota by balancing the abundance of most of the genera displaced by the Salmonella challenge with clearer effects observed with continuous supplementation of the probiotic.The levels of acetate and butyrate in the faeces were not affected(P>0.05)by Salmonella challenge and the butyrate level was increased by the continuous feeding of the probiotic.Probiotic supplementation in Salmonella challenged chickens resulted in higher level of propionate.Continuous probiotic supplementation decreased(P<0.05)the overall mean load of Salmonella in faeces and had a significant effect on Salmonella load reduction in internal organs.Conclusions:Salmonella challenge negatively impacts the diversity and abundance of many gut microbial genera involved in important functions such as organic acid and vitamin production.Strategic feeding of a Bacillus based probiotic helps in restoring many of the microbial genera displaced by Salmonella Typhimurium challenge.

Response of Heterotrophic Bacteria Abundance and Community Structure to Asian Dust Addition in the Oligotrophic Northwest Pacific Ocean

The dust storms from the continent usually affect the abundance and diversity of planktons by supplying trace elements. As such, the response of heterotrophic planktonic bacteria to dusts, nutrients(i.e., nitrogen and phosphorus) or ferrous dosages was investigated in the Kuroshio Extension region of the Northwest Pacific Ocean(NWPO) through on-board incubation experiments during an oceanographic survey in spring 2014. The flow cytometry and 16S rRNA high-throughput sequencing methods were applied to explore the abundance and community structure of bacteria, and the percentage of high nucleic acid bacteria(HNA%). The results showed that the heterotrophic bacteria abundance was low(average 2.55×10^5 cells mL^-1) and subjected to both nitrogen(N) and ferrous(Fe) limitation. Sand-dust deposition observably promoted the activity of heterotrophic planktonic bacteria. The maximum abundance of heterotrophic bacteria was 6.98×10^5 cells mL^-1 in the dust-dosage group, which was 44% higher than the control(P < 0.05). The HNA% in the dust-dosage group was 1.37 times higher than the control(P < 0.05). The activation mechanism was mainly related to the dissolution of N and Fe in the dusts. The relative abundance of genus Winogradskyella was significantly increased by dust deposition while the relative abundance of the genera Tenacibaculum and Hyphomonas was decreased. These variations of bacterial community structure were ascribed to the dissolution of nutrients N and P. Comparing the results of different experimental groups, this study concluded that dust storm improved the abundance of heterotrophic bacteria by dissolution of N and Fe.

Distribution of Bacterial Communities in Petroleum-Contaminated Soils from the Dagang Oilfield, China

Diversity in bacterial communities was investigated along a petroleum hydrocarbon content gradient(0-0.4043 g/g)in surface(5-10 cm)and subsurface(35-40 cm)petroleum-contaminated soil samples from the Dagang Oilfield,China.Using 16S rRNA Illumina high-throughput sequencing technology and several statistical methods,the bacterial diversity of the soil was studied.Subsequently,the environmental parameters were measured to analyze its relationship with the community variation.Nonmetric multidimensional scaling and analysis of similarities indicated a significant difference in the structure of the bacterial community between the nonpetroleum-contaminated surface and subsurface soils,but no differences were observed in different depths of petroleum-contaminated soil.Meanwhile,many significant correlations were obtained between diversity in soil bacterial community and physicochemical properties.Total petroleum hydrocarbon,total organic carbon,and total nitrogen were the three important factors that had the greatest impacts on the bacterial community distribution in the long-term petroleum-contaminated soils.Our research has provided references for the bacterial community distribution along a petroleum gradient in both surface and subsurface petroleum-contaminated soils of oilfield areas.

Ganjiang granule regulates cecal microflora and serum biochemical components in a rat model of constipation-predominant irritable bowel syndrome

Objective:To investigate the effects of Ganjiang granule(GG)on cecal microflora and serum biochemical components in rats with constipation-predominant irritable bowel syndrome(IBSeC).Methods:Twenty-four SpragueeDawley rats were randomly divided into four groups:control,model,GG,and probiotics.Rats in the model,GG,and probiotics groups received 3℃ tap water intragastrically;rats in the GG group were treated with GG;rats in the probiotics group were treated with probiotics.For all rats,enzyme-linked immunosorbent assay and colorimetry were used to assess serum biochemical components related to gastrointestinal function;16S rRNA high-throughput sequencing was used to analyze the cecal microflora.Results:The serum level of 5-hydroxytryptamine(HT)was higher in the model group than in the control group(Z=-2.082,P=.037).The model group exhibited changes in cecal microflora:the relative abundances of Lactobacillus decreased(Z=-2.882,P=.004)and Dorea increased(t=-3.030,P=.023),compared with the control group.The GG and probiotics groups exhibited normal serum levels of 5-HT.The GG and probiotics groups exhibited improved serum levels of gastrin;the probiotics group exhibited an improved serum level of vasoactive intestinal peptide.Compared with the model group,The GG group exhibited greater relative abundance of Ruminococcus(Z=-2.402,P=.016);the probiotics group exhibited greater relative abundance of SMB53(Z=-2.823,P=.005)and lower relative abundances of Desulfovibrio(Z=-2.823,P=.005)and Facklamia(Z=-2.608,P=.009).Conclusion:The effects of GG on IBS-C may be related to regulation of the serum level of 5-HT,as well as elevated relative abundance of beneficial bacteria in cecal microflora.

Polyphyly in 16S rRNA-based LVTree Versus Monophyly in Whole-genome-based CVTree

We report an important but long-overlooked manifestation of low-resolution power of 16S rRNA sequence analysis at the species level, namely, in 16S rRNA-based phylogenetic trees polyphyletic placements of closely-related species are abundant compared to those in genomebased phylogeny. This phenomenon makes the demarcation of genera within many families ambiguous in the 16S rRNA-based taxonomy. In this study, we reconstructed phylogenetic relationship for more than ten thousand prokaryote genomes using the CVTree method, which is based on wholegenome information. And many such genera, which are polyphyletic in 16S rRNA-based trees, are well resolved as monophyletic clusters by CVTree. We believe that with genome sequencing of prokaryotes becoming a commonplace, genome-based phylogeny is doomed to play a definitive role in the construction of a natural and objective taxonomy.

Assessing the microbiota of recycled bedding sand on a Wisconsin dairy farm

Background:Sand is often considered the preferred bedding material for dairy cows as it is thought to have lower bacterial counts than organic bedding materials and cows bedded on sand experience fewer cases of lameness and disease.Sand can also be efficiently recycled and reused,making it cost-effective.However,some studies have suggested that the residual organic material present in recycled sand can serve as a reservoir for commensal and pathogenic bacteria,although no studies have yet characterized the total bacterial community composition.Here we sought to characterize the bacterial community composition of a Wisconsin dairy farm bedding sand recycling system and its dynamics across several stages of the recycling process during both summer and winter using 16S rRNA gene amplicon sequencing.Results:Bacterial community compositions of the sand recycling system differed by both seasons and stage.Summer samples had higher richness and distinct community compositions,relative to winter samples.In both summer and winter samples,the diversity of recycled sand decreased with time drying in the recycling room.Compositionally,summer sand 14 d post-recycling was enriched in operational taxonomic units(OTUs)belonging to the genera Acinetobacter and Pseudomonas,relative to freshly washed sand and sand from cow pens.In contrast,no OTUs were found to be enriched in winter sand.The sand recycling system contained an overall core microbiota of 141 OTUs representing 68.45%±10.33%SD of the total bacterial relative abundance at each sampled stage.The 4 most abundant genera in this core microbiota included Acinetobacter,Psychrobacter,Corynebacterium,and Pseudomonas.Acinetobacter was present in greater abundance in summer samples,whereas Psychrobacter and Corynebacterium had higher relative abundances in winter samples.Pseudomonas had consistent relative abundances across both seasons.Conclusions:These findings highlight the potential of recycled bedding sand as a bacterial reservoir that warrants further study.

Determining the protective effects of Ma-Mu-Ran Antidiarrheal Capsules against acute DSS-induced enteritis using 16S rRNA gene sequencing and fecal metabolomics

Ma-Mu-Ran Antidiarrheal Capsules(MMRAC)is traditional Chinese medicine that has been used to treat diarrhea caused by acute enteritis(AE)and bacillary dysentery in Xinjiang(China)for many years.However,the potential therapeutic mechanism of MMRAC for AE and its regulatory mechanism on host metabolism is unclear.This study used fecal metabolomics profiling with GC/MS and 16S rRNA gene sequencing analysis to explore the potential regulatory mechanisms of MMRAC on a dextran sulfate sodium salt(DSS)-induced mouse model of AE.Fecal metabolomics-based analyses were performed to detect the differentially expressed metabolites and metabolic pathways.The 16S rRNA gene sequencing analysis was used to assess the altered gut microbes at the genus level and for functional prediction.Moreover,Pearson correlation analysis was used to integrate differentially expressed metabolites and altered bacterial genera.The results revealed that six intestinal bacteria and seven metabolites mediated metabolic disorders(i.e.,metabolism of amino acid,carbohydrate,cofactors and vitamins,and lipid)in AE mice.Besides,ten altered microbes mediated the differential expression of eight metabolites and regulated these metabolisms after MMRAC administration.Overall,these findings demonstrate that AE is associated with metabolic disorders and microbial dysbiosis.Further,we present that MMRAC exerts protective effects against AE by improving host metabolism through the intestinal flora.

Prevalence of Bacillus sp.among the biofilm forming community on Ti surface in marine environment

Prevalence of bacterial species involved in biomineralization of manganese on titanium （Ti） surfaces in marine environment was revealed in this research work. This study involves one year sea water exposure of Ti and their periodical biofilm characterization was carried out to quantify the manganese oxidizing bacterial （MOB） presence in the biofilm formed on titanium surfaces. The total viable count study of Ti coupons exposed to sea water for one year resulted in 60% of the MOB in overall biofilm population. The biochemical characterization of MOB isolates were performed for the genus level identifcation of the seven bacterial isolates. Further, the seven strains were subjected to 16S rRNA gene sequencing. Evolutionary analysis was performed using MEGA 7 to obtain closely related strains within the groups. The manganese oxidizing ability of the bacterial isolates were determined with Leucoberbelin Blue Assay （LBB） and Atomic Absorption Spectroscopy studies （hAS）. The results show that among the isolated marine MOB species, Bacillus sp. and Leptothrix sp. have the maximum Mn oxidizing property. The microtitre plate assay was performed to determine the biofilm forming ability of the isolated marine MOB species. All the results have confirmed the prevalence of Bacillus sp. among the biofilm colonizers on Ti surfaces when exposed in sea water.

Effects of enrichmemt planting with native tree species on bacterial community structure and potential impact on Eucalyptus plantations in southern China

Multi-generational planting of Eucalyptus species degrades soil quality but the introduction of legumes can improve soil fertility and microbial diversity.However,the effects of introducing non-legume native tree species on soil nutrients and bacterial community structure remain poorly understood.This study investigated the impacts of the conversion of third generation monoculture Eucalyptus plantations to mixed systems including Eucalyptus urograndis with Cinnamomum camphora(EC)and E.urograndis with Castanopsis hystrix(EH),on soil chemical and biochemical properties and bacterial community structure,diversity and functions.First generation E.urophylla plantations were the control.Results show that planting the third generation Eucalyptus led to a significant decrease in p H,organic matter,nutrient content,enzyme activities(invertin,acid phosphataes,and urease),and bacterialα-diversity compare to the controls.However,the mixed planting showed significant improvement in soil chemical and biochemical attributes and bacterialα-diversity,although the E.urograndis and C.hystrix planting had no improvement.Chloroflexi(oligotrophic bacteria)were significantly enriched in third generation Eucalyptus and Eucalyptus+C.hystrix,while proteobacteria increased significantly in the E.urograndis with C.camphora plantings.The relative abundance of multiple metabolic pathways increased significantly in the third generation Eucalyptus plantations whereas membrane transportrelated genes were enriched in soils of the mixed systems.The changes in bacterial community structures in the two mixed systems were driven by diversity,organic matter and acid phosphatase,while bacterial functions were affected by invertase,NO_(3)^(-)-N,diversity and urease.These results suggest that the transformation of successive monoculture Eucalyptus plantations into mixed plantations reduces the depletion of soil nutrients and enhances the ecological function of soil microorganisms.

Phenotypic and Genetic Characteristics of a Bordetella hinzii Isolated from a Rex Rabbit

A Bordetella hinzii strain, isolated from the lung of a dead breast-feeding infant rex rabbit with a respiratory infection, was used as study object to investigate its phenotypic and genetic characteristics. The strain could grow on MacConkey agar and was gram-negative, oxidase-positive, catalase-positive, short rodshaped, and it can produce alkali from malonate. Its 6S rRNA sequence was partially identical with B. hinzii （AF177667）. Animal experiments revealed nasal mucous hyperemia, tracheal congestion and bronchopneumonia in 28-day old rex rabbits. To sum up, this Bordetella hinzii isolate was deemed as a novel causative agent of respiratory disease in rex rabbit.