Enterotoxigenic E.coli is one of the bacterial pathogens contributing to the global resistance crisis in public health and animal husbandry.The problem of antibiotic resistance is becoming more and more serious,and ph...Enterotoxigenic E.coli is one of the bacterial pathogens contributing to the global resistance crisis in public health and animal husbandry.The problem of antibiotic resistance is becoming more and more serious,and phage is con-sidered one of the potential alternatives to antibiotics that could be utilized to treat bacterial infections.Our study isolated and identified a lytic phage PGX1 against multidrug-resistant enterotoxigenic E.coli EC6 strain from sew-age.The phage lysis profile revealed that PGX1 exhibited a lytic effect on multidrug-resistant enterotoxigenic E.coli strains of serotype O60.Through phage whole genome sequencing and bioinformatics analysis,PGX1 was found to be the class Caudoviricetes,family Autographiviridae,genus Teseptimavirus.The length of the PGX1 genome is about 37,009 bp,containing 54 open reading frames(ORFs).Notably,phage PGX1 lacks any lysogenic-related genes or virulence genes.Furthermore,phage PGX1 demonstrates strong adaptability,tolerance,and stability in various pH(pH4-10)and temperatures(4–40°C).The in vivo and in vitro tests demonstrated that phage PGX1 significantly removes and inhibits the formation of multidrug-resistant EC6 biofilm and effectively controls the Galleria mel-lonella larvae and enterotoxigenic E.coli EC6 during mice infection.In conclusion,the above findings demonstrated that phage PGX1 may be a novel antimicrobial agent to control multidrug-resistant E.coli infections.展开更多
Background Monoglycerides have emerged as a promising alternative to conventional practices due to their biolog-ical activities,including antimicrobial properties.However,few studies have assessed the efficacy of mono...Background Monoglycerides have emerged as a promising alternative to conventional practices due to their biolog-ical activities,including antimicrobial properties.However,few studies have assessed the efficacy of monoglyceride blend on weaned pigs and their impacts on performance,immune response,and gut health using a disease chal-lenge model.Therefore,this study aimed to investigate the effects of dietary monoglycerides of short-and medium-chain fatty acids on the immunity and gut health of weaned pigs experimentally infected with an enterotoxigenic Escherichia coli F18.Results Pigs supplemented with high-dose zinc oxide(ZNO)had greater(P<0.05)growth performance than other treatments,but no difference was observed in average daily feed intake between ZNO and monoglycerides groups during the post-challenge period.Pigs in ZNO and antibiotic groups had lower(P<0.05)severity of diarrhea than control,but the severity of diarrhea was not different between antibiotic and monoglycerides groups.Pigs fed with monoglycerides or ZNO had lower(P<0.05)serum haptoglobin on d 2 or 5 post-inoculation than control.Pigs in ZNO had greater(P<0.05)goblet cell numbers per villus,villus area and height,and villus height:crypt depth ratio(VH:CD)in duodenum on d 5 post-inoculation than pigs in other treatments.Pigs supplemented with monoglyc-erides,ZNO,or antibiotics had reduced(P<0.05)ileal crypt depth compared with control on d 5 post-inoculation,contributing to the increase(P=0.06)in VH:CD.Consistently,pigs in ZNO expressed the lowest(P<0.05)TNFa,IL6,IL10,IL12,IL1A,IL1B,and PTGS2 in ileal mucosa on d 5 post-inoculation,and no difference was observed in the expres-sion of those genes between ZNO and monoglycerides.Supplementation of ZNO and antibiotic had significant impacts on metabolic pathways in the serum compared with control,particularly on carbohydrate and amino acid metabolism,while limited impacts on serum metabolites were observed in monoglycerides group when compared with control.Conclusions The results suggest that supplementation of monoglyceride blend may enhance disease resist-ance of weaned pigs by alleviating the severity of diarrhea and mitigating intestinal and systemic inflammation,although the effectiveness may not be comparable to high-dose zinc oxide.展开更多
The gene cluster cfaABCED’ of enterotoxigenic Escherichia coli, encoding the fimbriae which is called colonization factor antigen located on a plasmid. It is positively regulated by cfaR, a member of the AraC family,...The gene cluster cfaABCED’ of enterotoxigenic Escherichia coli, encoding the fimbriae which is called colonization factor antigen located on a plasmid. It is positively regulated by cfaR, a member of the AraC family, and the cfaD’ gene region, which is located downstream of cfaE and is homologous to cfaR, had been described as a truncated cryptic gene. In the present study we observed that the CFA/ fimbriae subunit, cfaB, was expressed in lower amount by the cfaABCED’ clone pNTP513 in host E. coli HB101. The expression of CFA/ diminished by deletion of cfaD’ gene region from pNTP513, and was restored by acquisition of cfaD’ in trans. Furthermore, CFA/ expression by cfaD’ deletion mutant, the cfaABCE clone, was remarkably increased by the presence of cfaD’ in trans in a topoisomerase A deficient strain of E. coli DM800. These data suggest that cfaD’ region is a functional region of gene, that regulates the CFA/ expression with cfaR by unknown mechanism.展开更多
Background Infection with pathogenic bacteria during nonantibiotic breeding is one of the main causes of animal intestinal diseases.Oleanolic acid(OA)is a pentacyclic triterpene that is ubiquitous in plants.Our previo...Background Infection with pathogenic bacteria during nonantibiotic breeding is one of the main causes of animal intestinal diseases.Oleanolic acid(OA)is a pentacyclic triterpene that is ubiquitous in plants.Our previous work demonstrated the protective effect of OA on intestinal health,but the underlying molecular mechanisms remain unclear.This study investigated whether dietary supplementation with OA can prevent diarrhea and intestinal immune dysregulation caused by enterotoxigenic Escherichia coli(ETEC)in piglets.The key molecular role of bile acid receptor signaling in this process has also been explored.Results Our results demonstrated that OA supplementation alleviated the disturbance of bile acid metabolism in ETEC-infected piglets(P<0.05).OA supplementation stabilized the composition of the bile acid pool in piglets by regulating the enterohepatic circulation of bile acids and significantly increased the contents of UDCA and CDCA in the ileum and cecum(P<0.05).This may also explain why OA can maintain the stability of the intestinal microbiota structure in ETEC-challenged piglets.In addition,as a natural ligand of bile acid receptors,OA can reduce the severity of intestinal inflammation and enhance the strength of intestinal epithelial cell antimicrobial programs through the bile acid receptors TGR5 and FXR(P<0.05).Specifically,OA inhibited NF-κB-mediated intestinal inflammation by directly activating TGR5 and its downstream c AMP-PKA-CREB signaling pathway(P<0.05).Furthermore,OA enhanced CDCA-mediated MEK-ERK signaling in intestinal epithelial cells by upregulating the expression of FXR(P<0.05),thereby upregulating the expression of endogenous defense molecules in intestinal epithelial cells.Conclusions In conclusion,our findings suggest that OA-mediated regulation of bile acid metabolism plays an important role in the innate immune response,which provides a new diet-based intervention for intestinal diseases caused by pathogenic bacterial infections in piglets.展开更多
Background Our previous study has reported that supplementation of oligosaccharide-based polymer enhances gut health and disease resistance of pigs infected with enterotoxigenic E.coli(ETEC)F18 in a manner similar to ...Background Our previous study has reported that supplementation of oligosaccharide-based polymer enhances gut health and disease resistance of pigs infected with enterotoxigenic E.coli(ETEC)F18 in a manner similar to carbadox.The objective of this study was to investigate the impacts of oligosaccharide-based polymer or antibiotic on the host metabolic profiles and colon microbiota of weaned pigs experimentally infected with ETEC F18.Results Multivariate analysis highlighted the differences in the metabolic profiles of serum and colon digesta which were predominantly found between pigs supplemented with oligosaccharide-based polymer and antibiotic.The relative abundance of metabolic markers of immune responses and nutrient metabolisms,such as amino acids and carbohydrates,were significantly differentiated between the oligosaccharide-based polymer and antibiotic groups(q<0.2 and fold change>2.0).In addition,pigs in antibiotic had a reduced(P<0.05)relative abundance of Lachnospiraceae and Lactobacillaceae,whereas had greater(P<0.05)Clostridiaceae and Streptococcaceae in the colon digesta on d 11 post-inoculation(PI)compared with d 5 PI.Conclusions The impact of oligosaccharide-based polymer on the metabolic and microbial profiles of pigs is not fully understood,and further exploration is needed.However,current research suggest that various mechanisms are involved in the enhanced disease resistance and performance in ETEC-challenged pigs by supplementing this polymer.展开更多
Background:This study aimed to investigate the effects of oral administration of Enterococcus faecium NCIMB10415(E.faecium)on intestinal development,immunological parameters and gut microbiota of neonatal piglets chal...Background:This study aimed to investigate the effects of oral administration of Enterococcus faecium NCIMB10415(E.faecium)on intestinal development,immunological parameters and gut microbiota of neonatal piglets challenged with enterotoxigenic Escherichia coli K88(ETEC).A total of 961-day-old sow-reared piglets were randomly assigned to 2 groups,with 48 piglets in each group.The piglets were from 16 litters(6 piglets each litter),and 3 piglets each litter were allocated to the E.faecium-supplemented(PRO)group,while the other 3 piglets were allocated to the control(CON)group.After colostrum intake,piglets in the PRO group were orally administrated with 3×10~9 CFU E.faecium per day for a period of one week.On day 8,one piglet per litter from each group was challenged(CON+ETEC,PRO+ETEC)or not(CON-ETEC,PRO-ETEC)with ETEC in a 2×2 factorial arrangement of treatments.On day 10(2 days after challenge),blood and tissue samples were obtained from piglets.Results:Before ETEC challenge,there were no significant differences for the average daily gain(ADG)and fecal score between the two groups of piglets.After ETEC challenge,the challenged piglets had greater fecal score compared to the non-challenged piglets,whereas E.faecium administration was able to decrease the fecal score.Piglets challenged with ETEC had shorter villous height,deeper crypt depth,and reduced number of goblet cells in the jejunum and decreased m RNA abundance of claudin-1 in the ileum,whereas increased the percentage of lymphocytes,concentrations of IL-1βin the plasma and TNF-αin the ileal mucosa,as well as increased the m RNA abundances of innate immunity-related genes in the ileum tissue.These deleterious effects caused by ETEC were partly alleviated by feeding E.faecium.In addition,piglets in PRO-ETEC group had decreased the percentage of CD8^+T cells of the peripheral blood when compared to those in CON-ETEC group.Moreover,E.faecium administration increased Verrucomicrobia at phylum level and decreased Bilophila at genus level.Conclusions:These results suggest that oral administration of E.faecium alleviated the intestinal injury and diarrhea severity of neonatal piglets challenged by ETEC,partly through improving the intestinal microbiota and immune response.This offers a potential strategy of dietary intervention against intestinal impairment by ETEC in neonatal piglets.展开更多
Background Post-weaned piglets suffer from F18+Escherichia coli(E.coli)infections resulting in post-weaning diar-rhoea or oedema disease.Frequently used management strategies,including colistin and zinc oxide,have con...Background Post-weaned piglets suffer from F18+Escherichia coli(E.coli)infections resulting in post-weaning diar-rhoea or oedema disease.Frequently used management strategies,including colistin and zinc oxide,have contrib-uted to the emergence and spread of antimicrobial resistance.Novel antimicrobials capable of directly interacting with pathogens and modulating the host immune responses are being investigated.Lactoferrin has shown promising results against porcine enterotoxigenic E.coli strains,both in vitro and in vivo.Results We investigated the influence of bovine lactoferrin(bLF)on the microbiome of healthy and infected weaned piglets.Additionally,we assessed whether bLF influenced the immune responses upon Shiga toxin-producing E.coli(STEC)infection.Therefore,2 in vivo trials were conducted:a microbiome trial and a challenge infection trial,using an F18+STEC strain.BLF did not affect theα-andβ-diversity.However,bLF groups showed a higher relative abundance(RA)for the Actinobacteria phylum and the Bifidobacterium genus in the ileal mucosa.When analysing the immune response upon infection,the STEC group exhibited a significant increase in F18-specific IgG serum levels,whereas this response was absent in the bLF group.Conclusion Taken together,the oral administration of bLF did not have a notable impact on theα-andβ-diversity of the gut microbiome in weaned piglets.Nevertheless,it did increase the RA of the Actinobacteria phylum and Bifi-dobacterium genus,which have previously been shown to play an important role in maintaining gut homeostasis.Furthermore,bLF administration during STEC infection resulted in the absence of F18-specific serum IgG responses.展开更多
Pasteurellosis is the most prevalent, extremely contagious bacterial disease among domestic rabbits and is considered the leading cause of deaths in rabbits, resulting in enormous economic losses to the rabbit industr...Pasteurellosis is the most prevalent, extremely contagious bacterial disease among domestic rabbits and is considered the leading cause of deaths in rabbits, resulting in enormous economic losses to the rabbit industry. Screening for bacterial agents causing mortalities in rabbits revealed the presence of Enterobacteriacae species in approximately 42% of studied cases, with E. coli the most commonly isolated organism. The present study was designed to evaluate the immune response of rabbits vaccinated with a locally prepared, combined inactivated vaccine of Pasteurella multocida and E. coli, adjuvanated with Montanide ISA70. A total of 370 rabbits, aged 2 - 3 weeks, were divided into four groups: (G1) vaccinated with a polyvalent P. multocida vaccine, (G2) vaccinated with a polyvalent E. coli vaccine, (G3) vaccinated with a combined inactivated Montanide ISA70 vaccine of P. multocida and E. coli, and (G4) kept as a non-vaccinated control group. All rabbits received two doses of 0.5 ml of the prepared vaccines, administered one month apart, and were then challenged with virulent strains of P. multocida and E. coli three weeks after the second vaccination. The prepared vaccines were evaluated by determining humoral immunity using indirect haemagglutination (IHA) test and ELISA. The potency of the vaccines was assessed through challenge and determination of LD50. Experimental findings on the prepared polyvalent combined inactivated P. multocida and E. coli vaccine indicated that it is a potent vaccine, producing the highest antibody titers and a 90% protection rate against challenges with virulent strains of P. multocida type A, D2, and E. coli types O157, O151 and O125. Thus, this vaccine is promising in addressing both P. multocida and E. coli problems in rabbits, farms, providing significant protection, and we recommend its commercial production to help rabbit producers control these two major bacterial infections.展开更多
Background: Recently micro-organisms that synthesize extended-spectrum β-lactamase (ESBLs) were increased. The peculiarities of ESBL synthesis of Escherichia coli and Klebsiella pneumoniae strains that cause nosocomi...Background: Recently micro-organisms that synthesize extended-spectrum β-lactamase (ESBLs) were increased. The peculiarities of ESBL synthesis of Escherichia coli and Klebsiella pneumoniae strains that cause nosocomial urinary tract infections, surgical site infections and pneumonia in surgical clinic were studied. ESBL synthesis were observed 38.9% of E. coli strains obtained from urine, 92.3% of strains obtained from surgical site infections, and 50% of strains obtained from sputum. ESBL synthesis were observed 37.5% of K. pneumoniae strains obtained from urine, 85.7% of strains obtained from surgical site infections, and 60% of strains obtained from sputum. Different levels of ESBL synthesize of E. coli and K. pneumoniae strains isolated from different pattern is discussed. Conclusion. ESBL synthesis is common in E. coli and K. pneumoniae strains, which cause nosocomial infections. The frequency of occurrence of ESBL s synthesis among of these strains depends on clinical forms of nosocomial infections.展开更多
Background： This experiment was conducted to test the hypothesis that vitamin E（Vit E） and acetylsalicylic acid（ASA）, a cyclooxygenase-2（COX-2） inhibitor, will additively reduce the production of the immunosupp...Background： This experiment was conducted to test the hypothesis that vitamin E（Vit E） and acetylsalicylic acid（ASA）, a cyclooxygenase-2（COX-2） inhibitor, will additively reduce the production of the immunosuppressive molecule prostaglandin E_2（PGE_2） and hence reduce inflammatory responses in weaner pigs experimentally infected with an enterotoxigenic strain of E. coli.Methods： The experiment was conducted in a research facility with 192 individually-housed male weaner pigs（Landrace × Large White） weighing 6.6 ± 0.04 kg（mean ± SEM）. The pigs were experimentally infected with an enterotoxigenic strain of E. coli and were allocated to a 2 × 3 factorial design with the respective factors being without and with 125 ppm ASA and three levels of Vit E supplementation（50, 100 or 200 IU/kg diet, dl-α-tocopheryl acetate）.Results： Acetylsalicylic acid supplementation improved average daily gain（P 〈 0.05） and tended to improve feed：gain ratio（P 〈 0.10） during the first 14 d after weaning. Acetylsalicylic acid supplementation also improved（P 〈 0.001） amino acid utilization efficiency（as assessed by plasma urea level） and tended to decrease（P 〈 0.10） PGE2 production in the liver without affecting smal intestinal histology and tight junction protein mR NA expression in the jejunal epithelium. Vitamin E supplementation greater than 100 IU/kg diet sustained both the plasma Vit E concentration（P 〈 0.001） and plasma haptoglobin content（P 〈 0.001） after weaning. However, there was no additive effects of the combined supplementation of ASA and Vit E on performance, intestinal barrier function and inflammatory responses of weaned pigs.Conclusions： Although ASA and vitamin E improved amino acid utilization efficiency and reduced acute inflammatory responses, ASA and vitamin E did not additively reduce production of PGE2 and inflammatory responses in weaner pigs experimental y infected with an enterotoxigenic strain of E. coli.展开更多
Fecal coliform bacteria such as Escherichia coli (E. coli) are one of the main sources of groundwater pollution. An assessment of the transport and Persistence of E. coli in poultry litter amended Decatur silty Clay s...Fecal coliform bacteria such as Escherichia coli (E. coli) are one of the main sources of groundwater pollution. An assessment of the transport and Persistence of E. coli in poultry litter amended Decatur silty Clay soil and Hartsells Sandy soil was conducted using soil columns and simulated groundwater leaching. Enumeration of initial E. coli was determined to range from 2.851 × 10<sup>3</sup> to 3.044 × 10<sup>3</sup> CFU per gram of soil. These results have been used in a batch study to determine the persistence rate of E. coli in Decatur silty Clay soil and Hartsells Sandy soil. Results prove that E. coli survival growth rate increases for clay soil later than and at a higher rate than sandy soil. The column study has determined that E. coli was transported at a rate of 3.7 × 10<sup>6</sup><sup> </sup>CFU for Decatur silty loam and 6.3 × 10<sup>6</sup><sup> </sup>CFU for Hartsells sandy per gram of soil. Further, linear regression analysis predictions show higher porosity and soil moisture content affect transport, and Hartsells sandy soil has higher transport of E. coli due to its higher porosity and lower volumetric water content.展开更多
Honey has long been considered a wound treatment used to keep cuts and other epidermal injuries clean. This study tested that claim by comparing manuka honey used in medicine today, local unprocessed honey taken strai...Honey has long been considered a wound treatment used to keep cuts and other epidermal injuries clean. This study tested that claim by comparing manuka honey used in medicine today, local unprocessed honey taken straight from a hive, and pasteurized honey found at a store, on strains of E. coli and S. epidermidis. The study evaluated the effects these honeys had on bacterial growth to determine which had the greatest inhibition of bacterial growth. To determine this, plates streaked with strains of E. coli or S. epidermidis bacteria and agar wells filled with one of the honeys were incubated and subsequently the diameter of the zone of inhibition was measured. After 20 trials using each honey and bacteria type, manuka and unprocessed were shown to have a statistically significant advantage over the pasteurized honey at inhibiting the growth of E. coli and S. epidermidis, though it was variable whether manuka had an advantage over the unprocessed honey.展开更多
This study was designed to find the susceptibility of Nitrofurantoin and Fosfomycin among urinary isolates of Escherichia.coli.Four hundred(400)urine samples were collected for susceptibility of nitrofurantoin and fos...This study was designed to find the susceptibility of Nitrofurantoin and Fosfomycin among urinary isolates of Escherichia.coli.Four hundred(400)urine samples were collected for susceptibility of nitrofurantoin and fosfomycin among urinary isolates of E.coli.All indoor and outdoor patients'urinary samples yielded growth of E.coli.Mid-stream urine specimens were inoculated on blood agar and CLED agar and incubated at 35±2°C.Growth was observed,and Escherichia coli was identified by Gram staining,Catalase,Motility test and API 20E(Bio murex)as per standard procedure.Antimicrobial susceptibility testing of isolates for nitrofurantoin and fosfomycin was carried out by the modified Kirby-Bauer disc diffusion method according to CLSI guidelines ATCC 25922.E.coli was used as a quality control strain.A total of 400 samples were tested susceptibility of nitrofurantoin and fosfomycin among urinary isolates of E.coli during this period.A total of 400 samples yielded the growth of E.coli,out of which 178(44.5%)were male and 222(55.5%)were female samples.Among males,18(10%)were tolerant to nitrofurantoin,and 2(1.1%)were tolerant to fosfomycin.Among females,9(4.09%)were susceptible to nitrofurantoin while 6(2.72%)were susceptible to fosfomycin.Among age groups below 45 years old,6(4.76%)were tolerant to nitrofurantoin,and 2(1.58%)were sensitive to fosfomycin.Between 46-66 years old,4(2.81%)were sensitive to nitrofurantoin,and 3(2.11%)were sensitive to fosfomycin.Between 67-90 years old,17(12.87%)were sensitive to nitrofurantoin,and 4(3.03%)were tolerant to fosfomycin.Fosfomycin and nitrofurantoin showed good susceptibility in urinary isolates of E.coli and can be used empirically in our setup.展开更多
Escherichia coli MLB(MG1655ΔpflBΔldhA),which can hardly grow on glucose with little succinate accumulation under anaerobic conditions.Two-stage fermentation is a fermentation in which the first stage is used for cel...Escherichia coli MLB(MG1655ΔpflBΔldhA),which can hardly grow on glucose with little succinate accumulation under anaerobic conditions.Two-stage fermentation is a fermentation in which the first stage is used for cell growth and the second stage is used for product production.The ability of glucose consumption and succinate production of MLB under anaerobic conditions can be improved significantly by using acetate as the solo carbon source under aerobic condition during the two-stage fermentation.Then,the adaptive laboratory evolution(ALE)of growing on acetate was applied here.We assumed that the activities of succinate production related enzymes might be further improved in this study.E.coli MLB46-05 evolved from MLB and it had an improved growth phenotype on acetate.Interestingly,in MLB46-05,the yield and tolerance of succinic acid in the anaerobic condition of two-stage fermentation were improved significantly.According to transcriptome analysis,upregulation of the glyoxylate cycle and the activity of stress regulatory factors are the possible reasons for the elevated yield.And the increased tolerance to acetate made it more tolerant to high concentrations of glucose and succinate.Finally,strain MLB46-05 produced 111 g/L of succinic acid with a product yield of 0.74 g/g glucose.展开更多
Defining suitable enzymes for reaction steps in novel synthetic pathways is crucial for developing microbial cell factories for non-natural products.Here,we developed a computational workflow to identify C12 alcohol-a...Defining suitable enzymes for reaction steps in novel synthetic pathways is crucial for developing microbial cell factories for non-natural products.Here,we developed a computational workflow to identify C12 alcohol-active UDP-glycosyltransferases.The workflow involved three steps:(1)assembling initial candidates of putative UDP-glycosyltransferases,(2)refining selection by examining conserved regions,and(3)3D structure prediction and molecular docking.Genomic sequences from Candida,Pichia,Rhizopus,and Thermotoga,known for lauryl glucoside synthesis via whole-cell biocatalysis,were screened.Out of 240 predicted glycosyltransferases,8 candidates annotated as glycosyltransferases were selected after filtering out those with signal peptides and identifying conserved UDP-glycosyltransferase regions.These proteins underwent 3D structure prediction and molecular docking with 1-dodecanol.RO3G,a candidate from Rhizopus delemar RA 99-880 with a relatively high ChemPLP fitness score,was selected and expressed in Escherichia coli BL21(DE3).It was further characterized using a feeding experiment with 1-dodecanol.Results confirmed that the RO3G-expressing strain could convert 1-dodecanol to lauryl glucoside,as quantified by HPLC and identified by targeted LC-MS.Monitoring the growth and fermentation profiles of the engineered strain revealed that RO3G expression did not affect cell growth.Interestingly,acetate,a major fermentation product,was reduced in the RO3G-expressing strain compared to the GFP-expressing strain,suggesting a redirection of flux from acetate to other pathways.Overall,this work presents a successful workflow for discovering UDP-glycosyltransferase enzymes with confirmed activity toward 1-dodecanol for lauryl glucoside production.展开更多
Enterotoxigenic Escherichio coli （ETEC） causes neonatal and post-weaning diarrhea in pigs. In order to determine the risk factors, rectal/fecal swabs and visceral organs obtained from pig farms in two regions of Sou...Enterotoxigenic Escherichio coli （ETEC） causes neonatal and post-weaning diarrhea in pigs. In order to determine the risk factors, rectal/fecal swabs and visceral organs obtained from pig farms in two regions of South Africa were analyzed microbiologically against risk variables. Seventy-two percent of young pigs were found to be positive for ETEC toxin genes; estB （38.9%）, estB/STAP （25%）, and estB/LT （13.9%） were dominant. Risk factors for ETEC-diarrhea in pigs include： leaving sick piglets in a pen with healthy piglets [odds ratio （OR） = 33.52; P 〈 0.0001]; water spillage in pen （OR = 42.87; P 〈 0.0001）; hypothermic piglets （OR = 7.29; P 〈 0.0001）; runt piglets in pen with healthy littermates （OR = 3.65; P 〈 0.0001）; and prolonged use of antibiotics （OR = 3.05; P = 0.05）.展开更多
Background:The forest musk deer,a rare fauna species found in China,is famous for its musk secretion which is used in selected Traditional Chinese medicines.However,over-hunting has led to musk deer becoming an endang...Background:The forest musk deer,a rare fauna species found in China,is famous for its musk secretion which is used in selected Traditional Chinese medicines.However,over-hunting has led to musk deer becoming an endangered species,and their survival is also greatly challenged by various high incidence and high mortality respiratory and intestinal diseases such as septic pneumonia and enteritis.Accumulating evidence has demonstrated that Akkermannia muciniphila(AKK)is a promising probiotic,and we wondered whether AKK could be used as a food additive in animal breeding pro-grammes to help prevent intestinal diseases.Methods:We isolated one AKK strain from musk deer feces(AKK-D)using an im-proved enrichment medium combined with real-time PCR.After confirmation by 16S rRNA gene sequencing,a series of in vitro tests was conducted to evaluate the probiotic effects of AKK-D by assessing its reproductive capability,simulated gas-trointestinal fluid tolerance,acid and bile salt resistance,self-aggregation ability,hy-drophobicity,antibiotic sensitivity,hemolysis,harmful metabolite production,biofilm formation ability,and bacterial adhesion to gastrointestinal mucosa.Results:The AKK-D strain has a probiotic function similar to that of the standard strain in humans(AKK-H).An in vivo study found that AKK-D significantly amelio-rated symptoms in the enterotoxigenic Escherichia coli(ETEC)-induced murine diar-rhea model.AKK-D improved organ damage,inhibited inflammatory responses,and improved intestinal barrier permeability.Additionally,AKK-D promoted the reconsti-tution and maintenance of the homeostasis of gut microflora,as indicated by the fact that AKK-D-treated mice showed a decrease in Bacteroidetes and an increase in the proportion of other beneficial bacteria like Muribaculaceae,Muribaculum,and unclas-sified f_Lachnospiaceae compared with the diarrhea model mice.Conclusion:Taken together,our data show that this novel AKK-D strain might be a potential probiotic for use in musk deer breeding,although further extensive system-atic research is still needed.展开更多
Several technical parameters were studied during the fermentation of recombinant E.coli for the production of collagen-like biopolymer.The effects of dissolved oxygen as well as glucose concentration on fermentation w...Several technical parameters were studied during the fermentation of recombinant E.coli for the production of collagen-like biopolymer.The effects of dissolved oxygen as well as glucose concentration on fermentation were observed.The OD 600 value could reach 98 when dissolved oxygen was controlled at 50% and glucose around 1%.The production of human-like collagen with a yield of 29.4% was obtained.展开更多
基金supported by grants from the National Program on Key Research Project of China[2022YFD1800800,2021YFD1800300]the Yingzi Tech&Huazhong Agricultural University Intelligent Research Institute of Food Health[No.IRIFH202209,No.IRIFH202301]The National Program on Key Research Project of China,2022YFD1800800,Ping Qian,2021YFD1800300,Ping Qian,The Yingzi Tech&Huazhong Agricultural University Intelligent Research Institute of Food Health,IRIFH202209,Ping Qian,IRIFH202301,Ping Qian.
文摘Enterotoxigenic E.coli is one of the bacterial pathogens contributing to the global resistance crisis in public health and animal husbandry.The problem of antibiotic resistance is becoming more and more serious,and phage is con-sidered one of the potential alternatives to antibiotics that could be utilized to treat bacterial infections.Our study isolated and identified a lytic phage PGX1 against multidrug-resistant enterotoxigenic E.coli EC6 strain from sew-age.The phage lysis profile revealed that PGX1 exhibited a lytic effect on multidrug-resistant enterotoxigenic E.coli strains of serotype O60.Through phage whole genome sequencing and bioinformatics analysis,PGX1 was found to be the class Caudoviricetes,family Autographiviridae,genus Teseptimavirus.The length of the PGX1 genome is about 37,009 bp,containing 54 open reading frames(ORFs).Notably,phage PGX1 lacks any lysogenic-related genes or virulence genes.Furthermore,phage PGX1 demonstrates strong adaptability,tolerance,and stability in various pH(pH4-10)and temperatures(4–40°C).The in vivo and in vitro tests demonstrated that phage PGX1 significantly removes and inhibits the formation of multidrug-resistant EC6 biofilm and effectively controls the Galleria mel-lonella larvae and enterotoxigenic E.coli EC6 during mice infection.In conclusion,the above findings demonstrated that phage PGX1 may be a novel antimicrobial agent to control multidrug-resistant E.coli infections.
文摘Background Monoglycerides have emerged as a promising alternative to conventional practices due to their biolog-ical activities,including antimicrobial properties.However,few studies have assessed the efficacy of monoglyceride blend on weaned pigs and their impacts on performance,immune response,and gut health using a disease chal-lenge model.Therefore,this study aimed to investigate the effects of dietary monoglycerides of short-and medium-chain fatty acids on the immunity and gut health of weaned pigs experimentally infected with an enterotoxigenic Escherichia coli F18.Results Pigs supplemented with high-dose zinc oxide(ZNO)had greater(P<0.05)growth performance than other treatments,but no difference was observed in average daily feed intake between ZNO and monoglycerides groups during the post-challenge period.Pigs in ZNO and antibiotic groups had lower(P<0.05)severity of diarrhea than control,but the severity of diarrhea was not different between antibiotic and monoglycerides groups.Pigs fed with monoglycerides or ZNO had lower(P<0.05)serum haptoglobin on d 2 or 5 post-inoculation than control.Pigs in ZNO had greater(P<0.05)goblet cell numbers per villus,villus area and height,and villus height:crypt depth ratio(VH:CD)in duodenum on d 5 post-inoculation than pigs in other treatments.Pigs supplemented with monoglyc-erides,ZNO,or antibiotics had reduced(P<0.05)ileal crypt depth compared with control on d 5 post-inoculation,contributing to the increase(P=0.06)in VH:CD.Consistently,pigs in ZNO expressed the lowest(P<0.05)TNFa,IL6,IL10,IL12,IL1A,IL1B,and PTGS2 in ileal mucosa on d 5 post-inoculation,and no difference was observed in the expres-sion of those genes between ZNO and monoglycerides.Supplementation of ZNO and antibiotic had significant impacts on metabolic pathways in the serum compared with control,particularly on carbohydrate and amino acid metabolism,while limited impacts on serum metabolites were observed in monoglycerides group when compared with control.Conclusions The results suggest that supplementation of monoglyceride blend may enhance disease resist-ance of weaned pigs by alleviating the severity of diarrhea and mitigating intestinal and systemic inflammation,although the effectiveness may not be comparable to high-dose zinc oxide.
文摘The gene cluster cfaABCED’ of enterotoxigenic Escherichia coli, encoding the fimbriae which is called colonization factor antigen located on a plasmid. It is positively regulated by cfaR, a member of the AraC family, and the cfaD’ gene region, which is located downstream of cfaE and is homologous to cfaR, had been described as a truncated cryptic gene. In the present study we observed that the CFA/ fimbriae subunit, cfaB, was expressed in lower amount by the cfaABCED’ clone pNTP513 in host E. coli HB101. The expression of CFA/ diminished by deletion of cfaD’ gene region from pNTP513, and was restored by acquisition of cfaD’ in trans. Furthermore, CFA/ expression by cfaD’ deletion mutant, the cfaABCE clone, was remarkably increased by the presence of cfaD’ in trans in a topoisomerase A deficient strain of E. coli DM800. These data suggest that cfaD’ region is a functional region of gene, that regulates the CFA/ expression with cfaR by unknown mechanism.
基金financially supported by the National Natural Science Foundation of China(Grant No.31972580 and U21A20252)the China Agriculture Research System(CARS-35)+1 种基金the Science Fund for Distinguished Young Scholars of Heilongjiang Province(JQ2022C002)the Support Project of Young Leading Talents of Northeast Agricultural University(NEAU2023QNLJ-017)。
文摘Background Infection with pathogenic bacteria during nonantibiotic breeding is one of the main causes of animal intestinal diseases.Oleanolic acid(OA)is a pentacyclic triterpene that is ubiquitous in plants.Our previous work demonstrated the protective effect of OA on intestinal health,but the underlying molecular mechanisms remain unclear.This study investigated whether dietary supplementation with OA can prevent diarrhea and intestinal immune dysregulation caused by enterotoxigenic Escherichia coli(ETEC)in piglets.The key molecular role of bile acid receptor signaling in this process has also been explored.Results Our results demonstrated that OA supplementation alleviated the disturbance of bile acid metabolism in ETEC-infected piglets(P<0.05).OA supplementation stabilized the composition of the bile acid pool in piglets by regulating the enterohepatic circulation of bile acids and significantly increased the contents of UDCA and CDCA in the ileum and cecum(P<0.05).This may also explain why OA can maintain the stability of the intestinal microbiota structure in ETEC-challenged piglets.In addition,as a natural ligand of bile acid receptors,OA can reduce the severity of intestinal inflammation and enhance the strength of intestinal epithelial cell antimicrobial programs through the bile acid receptors TGR5 and FXR(P<0.05).Specifically,OA inhibited NF-κB-mediated intestinal inflammation by directly activating TGR5 and its downstream c AMP-PKA-CREB signaling pathway(P<0.05).Furthermore,OA enhanced CDCA-mediated MEK-ERK signaling in intestinal epithelial cells by upregulating the expression of FXR(P<0.05),thereby upregulating the expression of endogenous defense molecules in intestinal epithelial cells.Conclusions In conclusion,our findings suggest that OA-mediated regulation of bile acid metabolism plays an important role in the innate immune response,which provides a new diet-based intervention for intestinal diseases caused by pathogenic bacterial infections in piglets.
基金supported by Pancosma SA,Geneva,Switzerland,Jastro & Shields Graduate Research Awardthe United States Department of Agriculture (USDA) National Institute of Food and Agriculture (NIFA),multistate projects W4002 and NC1202
文摘Background Our previous study has reported that supplementation of oligosaccharide-based polymer enhances gut health and disease resistance of pigs infected with enterotoxigenic E.coli(ETEC)F18 in a manner similar to carbadox.The objective of this study was to investigate the impacts of oligosaccharide-based polymer or antibiotic on the host metabolic profiles and colon microbiota of weaned pigs experimentally infected with ETEC F18.Results Multivariate analysis highlighted the differences in the metabolic profiles of serum and colon digesta which were predominantly found between pigs supplemented with oligosaccharide-based polymer and antibiotic.The relative abundance of metabolic markers of immune responses and nutrient metabolisms,such as amino acids and carbohydrates,were significantly differentiated between the oligosaccharide-based polymer and antibiotic groups(q<0.2 and fold change>2.0).In addition,pigs in antibiotic had a reduced(P<0.05)relative abundance of Lachnospiraceae and Lactobacillaceae,whereas had greater(P<0.05)Clostridiaceae and Streptococcaceae in the colon digesta on d 11 post-inoculation(PI)compared with d 5 PI.Conclusions The impact of oligosaccharide-based polymer on the metabolic and microbial profiles of pigs is not fully understood,and further exploration is needed.However,current research suggest that various mechanisms are involved in the enhanced disease resistance and performance in ETEC-challenged pigs by supplementing this polymer.
基金supported by the Projects of The National Key Research and Development Program of China(grant number 2016YFD0501204)Sichuan provincial project on S&T application and demonstration(grant number2016CC0070)the project on commercialization of research findings under funding of government of Sichuan province(grant number16ZHSF0385).
文摘Background:This study aimed to investigate the effects of oral administration of Enterococcus faecium NCIMB10415(E.faecium)on intestinal development,immunological parameters and gut microbiota of neonatal piglets challenged with enterotoxigenic Escherichia coli K88(ETEC).A total of 961-day-old sow-reared piglets were randomly assigned to 2 groups,with 48 piglets in each group.The piglets were from 16 litters(6 piglets each litter),and 3 piglets each litter were allocated to the E.faecium-supplemented(PRO)group,while the other 3 piglets were allocated to the control(CON)group.After colostrum intake,piglets in the PRO group were orally administrated with 3×10~9 CFU E.faecium per day for a period of one week.On day 8,one piglet per litter from each group was challenged(CON+ETEC,PRO+ETEC)or not(CON-ETEC,PRO-ETEC)with ETEC in a 2×2 factorial arrangement of treatments.On day 10(2 days after challenge),blood and tissue samples were obtained from piglets.Results:Before ETEC challenge,there were no significant differences for the average daily gain(ADG)and fecal score between the two groups of piglets.After ETEC challenge,the challenged piglets had greater fecal score compared to the non-challenged piglets,whereas E.faecium administration was able to decrease the fecal score.Piglets challenged with ETEC had shorter villous height,deeper crypt depth,and reduced number of goblet cells in the jejunum and decreased m RNA abundance of claudin-1 in the ileum,whereas increased the percentage of lymphocytes,concentrations of IL-1βin the plasma and TNF-αin the ileal mucosa,as well as increased the m RNA abundances of innate immunity-related genes in the ileum tissue.These deleterious effects caused by ETEC were partly alleviated by feeding E.faecium.In addition,piglets in PRO-ETEC group had decreased the percentage of CD8^+T cells of the peripheral blood when compared to those in CON-ETEC group.Moreover,E.faecium administration increased Verrucomicrobia at phylum level and decreased Bilophila at genus level.Conclusions:These results suggest that oral administration of E.faecium alleviated the intestinal injury and diarrhea severity of neonatal piglets challenged by ETEC,partly through improving the intestinal microbiota and immune response.This offers a potential strategy of dietary intervention against intestinal impairment by ETEC in neonatal piglets.
基金The research that yielded these results,was funded by the Belgian Federal Public Service of Health,Food Chain Safety and Environment through the contract RF 17/6314 LactoPigHealthMatthias Dierick is supported by the Flemish fund for scientific research(FWO3S036319).
文摘Background Post-weaned piglets suffer from F18+Escherichia coli(E.coli)infections resulting in post-weaning diar-rhoea or oedema disease.Frequently used management strategies,including colistin and zinc oxide,have contrib-uted to the emergence and spread of antimicrobial resistance.Novel antimicrobials capable of directly interacting with pathogens and modulating the host immune responses are being investigated.Lactoferrin has shown promising results against porcine enterotoxigenic E.coli strains,both in vitro and in vivo.Results We investigated the influence of bovine lactoferrin(bLF)on the microbiome of healthy and infected weaned piglets.Additionally,we assessed whether bLF influenced the immune responses upon Shiga toxin-producing E.coli(STEC)infection.Therefore,2 in vivo trials were conducted:a microbiome trial and a challenge infection trial,using an F18+STEC strain.BLF did not affect theα-andβ-diversity.However,bLF groups showed a higher relative abundance(RA)for the Actinobacteria phylum and the Bifidobacterium genus in the ileal mucosa.When analysing the immune response upon infection,the STEC group exhibited a significant increase in F18-specific IgG serum levels,whereas this response was absent in the bLF group.Conclusion Taken together,the oral administration of bLF did not have a notable impact on theα-andβ-diversity of the gut microbiome in weaned piglets.Nevertheless,it did increase the RA of the Actinobacteria phylum and Bifi-dobacterium genus,which have previously been shown to play an important role in maintaining gut homeostasis.Furthermore,bLF administration during STEC infection resulted in the absence of F18-specific serum IgG responses.
文摘Pasteurellosis is the most prevalent, extremely contagious bacterial disease among domestic rabbits and is considered the leading cause of deaths in rabbits, resulting in enormous economic losses to the rabbit industry. Screening for bacterial agents causing mortalities in rabbits revealed the presence of Enterobacteriacae species in approximately 42% of studied cases, with E. coli the most commonly isolated organism. The present study was designed to evaluate the immune response of rabbits vaccinated with a locally prepared, combined inactivated vaccine of Pasteurella multocida and E. coli, adjuvanated with Montanide ISA70. A total of 370 rabbits, aged 2 - 3 weeks, were divided into four groups: (G1) vaccinated with a polyvalent P. multocida vaccine, (G2) vaccinated with a polyvalent E. coli vaccine, (G3) vaccinated with a combined inactivated Montanide ISA70 vaccine of P. multocida and E. coli, and (G4) kept as a non-vaccinated control group. All rabbits received two doses of 0.5 ml of the prepared vaccines, administered one month apart, and were then challenged with virulent strains of P. multocida and E. coli three weeks after the second vaccination. The prepared vaccines were evaluated by determining humoral immunity using indirect haemagglutination (IHA) test and ELISA. The potency of the vaccines was assessed through challenge and determination of LD50. Experimental findings on the prepared polyvalent combined inactivated P. multocida and E. coli vaccine indicated that it is a potent vaccine, producing the highest antibody titers and a 90% protection rate against challenges with virulent strains of P. multocida type A, D2, and E. coli types O157, O151 and O125. Thus, this vaccine is promising in addressing both P. multocida and E. coli problems in rabbits, farms, providing significant protection, and we recommend its commercial production to help rabbit producers control these two major bacterial infections.
文摘Background: Recently micro-organisms that synthesize extended-spectrum β-lactamase (ESBLs) were increased. The peculiarities of ESBL synthesis of Escherichia coli and Klebsiella pneumoniae strains that cause nosocomial urinary tract infections, surgical site infections and pneumonia in surgical clinic were studied. ESBL synthesis were observed 38.9% of E. coli strains obtained from urine, 92.3% of strains obtained from surgical site infections, and 50% of strains obtained from sputum. ESBL synthesis were observed 37.5% of K. pneumoniae strains obtained from urine, 85.7% of strains obtained from surgical site infections, and 60% of strains obtained from sputum. Different levels of ESBL synthesize of E. coli and K. pneumoniae strains isolated from different pattern is discussed. Conclusion. ESBL synthesis is common in E. coli and K. pneumoniae strains, which cause nosocomial infections. The frequency of occurrence of ESBL s synthesis among of these strains depends on clinical forms of nosocomial infections.
基金support by Australian Cooperative Research Centre for High Integrity Australian Pork(Award number 2C-110 1213)
文摘Background： This experiment was conducted to test the hypothesis that vitamin E（Vit E） and acetylsalicylic acid（ASA）, a cyclooxygenase-2（COX-2） inhibitor, will additively reduce the production of the immunosuppressive molecule prostaglandin E_2（PGE_2） and hence reduce inflammatory responses in weaner pigs experimentally infected with an enterotoxigenic strain of E. coli.Methods： The experiment was conducted in a research facility with 192 individually-housed male weaner pigs（Landrace × Large White） weighing 6.6 ± 0.04 kg（mean ± SEM）. The pigs were experimentally infected with an enterotoxigenic strain of E. coli and were allocated to a 2 × 3 factorial design with the respective factors being without and with 125 ppm ASA and three levels of Vit E supplementation（50, 100 or 200 IU/kg diet, dl-α-tocopheryl acetate）.Results： Acetylsalicylic acid supplementation improved average daily gain（P 〈 0.05） and tended to improve feed：gain ratio（P 〈 0.10） during the first 14 d after weaning. Acetylsalicylic acid supplementation also improved（P 〈 0.001） amino acid utilization efficiency（as assessed by plasma urea level） and tended to decrease（P 〈 0.10） PGE2 production in the liver without affecting smal intestinal histology and tight junction protein mR NA expression in the jejunal epithelium. Vitamin E supplementation greater than 100 IU/kg diet sustained both the plasma Vit E concentration（P 〈 0.001） and plasma haptoglobin content（P 〈 0.001） after weaning. However, there was no additive effects of the combined supplementation of ASA and Vit E on performance, intestinal barrier function and inflammatory responses of weaned pigs.Conclusions： Although ASA and vitamin E improved amino acid utilization efficiency and reduced acute inflammatory responses, ASA and vitamin E did not additively reduce production of PGE2 and inflammatory responses in weaner pigs experimental y infected with an enterotoxigenic strain of E. coli.
文摘Fecal coliform bacteria such as Escherichia coli (E. coli) are one of the main sources of groundwater pollution. An assessment of the transport and Persistence of E. coli in poultry litter amended Decatur silty Clay soil and Hartsells Sandy soil was conducted using soil columns and simulated groundwater leaching. Enumeration of initial E. coli was determined to range from 2.851 × 10<sup>3</sup> to 3.044 × 10<sup>3</sup> CFU per gram of soil. These results have been used in a batch study to determine the persistence rate of E. coli in Decatur silty Clay soil and Hartsells Sandy soil. Results prove that E. coli survival growth rate increases for clay soil later than and at a higher rate than sandy soil. The column study has determined that E. coli was transported at a rate of 3.7 × 10<sup>6</sup><sup> </sup>CFU for Decatur silty loam and 6.3 × 10<sup>6</sup><sup> </sup>CFU for Hartsells sandy per gram of soil. Further, linear regression analysis predictions show higher porosity and soil moisture content affect transport, and Hartsells sandy soil has higher transport of E. coli due to its higher porosity and lower volumetric water content.
文摘Honey has long been considered a wound treatment used to keep cuts and other epidermal injuries clean. This study tested that claim by comparing manuka honey used in medicine today, local unprocessed honey taken straight from a hive, and pasteurized honey found at a store, on strains of E. coli and S. epidermidis. The study evaluated the effects these honeys had on bacterial growth to determine which had the greatest inhibition of bacterial growth. To determine this, plates streaked with strains of E. coli or S. epidermidis bacteria and agar wells filled with one of the honeys were incubated and subsequently the diameter of the zone of inhibition was measured. After 20 trials using each honey and bacteria type, manuka and unprocessed were shown to have a statistically significant advantage over the pasteurized honey at inhibiting the growth of E. coli and S. epidermidis, though it was variable whether manuka had an advantage over the unprocessed honey.
文摘This study was designed to find the susceptibility of Nitrofurantoin and Fosfomycin among urinary isolates of Escherichia.coli.Four hundred(400)urine samples were collected for susceptibility of nitrofurantoin and fosfomycin among urinary isolates of E.coli.All indoor and outdoor patients'urinary samples yielded growth of E.coli.Mid-stream urine specimens were inoculated on blood agar and CLED agar and incubated at 35±2°C.Growth was observed,and Escherichia coli was identified by Gram staining,Catalase,Motility test and API 20E(Bio murex)as per standard procedure.Antimicrobial susceptibility testing of isolates for nitrofurantoin and fosfomycin was carried out by the modified Kirby-Bauer disc diffusion method according to CLSI guidelines ATCC 25922.E.coli was used as a quality control strain.A total of 400 samples were tested susceptibility of nitrofurantoin and fosfomycin among urinary isolates of E.coli during this period.A total of 400 samples yielded the growth of E.coli,out of which 178(44.5%)were male and 222(55.5%)were female samples.Among males,18(10%)were tolerant to nitrofurantoin,and 2(1.1%)were tolerant to fosfomycin.Among females,9(4.09%)were susceptible to nitrofurantoin while 6(2.72%)were susceptible to fosfomycin.Among age groups below 45 years old,6(4.76%)were tolerant to nitrofurantoin,and 2(1.58%)were sensitive to fosfomycin.Between 46-66 years old,4(2.81%)were sensitive to nitrofurantoin,and 3(2.11%)were sensitive to fosfomycin.Between 67-90 years old,17(12.87%)were sensitive to nitrofurantoin,and 4(3.03%)were tolerant to fosfomycin.Fosfomycin and nitrofurantoin showed good susceptibility in urinary isolates of E.coli and can be used empirically in our setup.
基金supported by the National Key R&D Program of China(2021YFC2101300)Science and Technology Commission of Shanghai Municipality(21DZ1209100)Partially supported by Open Funding Project of the State Key Laboratory of Bioreactor Engineering.
文摘Escherichia coli MLB(MG1655ΔpflBΔldhA),which can hardly grow on glucose with little succinate accumulation under anaerobic conditions.Two-stage fermentation is a fermentation in which the first stage is used for cell growth and the second stage is used for product production.The ability of glucose consumption and succinate production of MLB under anaerobic conditions can be improved significantly by using acetate as the solo carbon source under aerobic condition during the two-stage fermentation.Then,the adaptive laboratory evolution(ALE)of growing on acetate was applied here.We assumed that the activities of succinate production related enzymes might be further improved in this study.E.coli MLB46-05 evolved from MLB and it had an improved growth phenotype on acetate.Interestingly,in MLB46-05,the yield and tolerance of succinic acid in the anaerobic condition of two-stage fermentation were improved significantly.According to transcriptome analysis,upregulation of the glyoxylate cycle and the activity of stress regulatory factors are the possible reasons for the elevated yield.And the increased tolerance to acetate made it more tolerant to high concentrations of glucose and succinate.Finally,strain MLB46-05 produced 111 g/L of succinic acid with a product yield of 0.74 g/g glucose.
基金This work(Grant No.RGNS 64-069)was financially supported by Office of the Permanent Secretary,Ministry of Higher Education,Science,Research and Innovationpartially supported by Chiang Mai University.
文摘Defining suitable enzymes for reaction steps in novel synthetic pathways is crucial for developing microbial cell factories for non-natural products.Here,we developed a computational workflow to identify C12 alcohol-active UDP-glycosyltransferases.The workflow involved three steps:(1)assembling initial candidates of putative UDP-glycosyltransferases,(2)refining selection by examining conserved regions,and(3)3D structure prediction and molecular docking.Genomic sequences from Candida,Pichia,Rhizopus,and Thermotoga,known for lauryl glucoside synthesis via whole-cell biocatalysis,were screened.Out of 240 predicted glycosyltransferases,8 candidates annotated as glycosyltransferases were selected after filtering out those with signal peptides and identifying conserved UDP-glycosyltransferase regions.These proteins underwent 3D structure prediction and molecular docking with 1-dodecanol.RO3G,a candidate from Rhizopus delemar RA 99-880 with a relatively high ChemPLP fitness score,was selected and expressed in Escherichia coli BL21(DE3).It was further characterized using a feeding experiment with 1-dodecanol.Results confirmed that the RO3G-expressing strain could convert 1-dodecanol to lauryl glucoside,as quantified by HPLC and identified by targeted LC-MS.Monitoring the growth and fermentation profiles of the engineered strain revealed that RO3G expression did not affect cell growth.Interestingly,acetate,a major fermentation product,was reduced in the RO3G-expressing strain compared to the GFP-expressing strain,suggesting a redirection of flux from acetate to other pathways.Overall,this work presents a successful workflow for discovering UDP-glycosyltransferase enzymes with confirmed activity toward 1-dodecanol for lauryl glucoside production.
文摘Enterotoxigenic Escherichio coli （ETEC） causes neonatal and post-weaning diarrhea in pigs. In order to determine the risk factors, rectal/fecal swabs and visceral organs obtained from pig farms in two regions of South Africa were analyzed microbiologically against risk variables. Seventy-two percent of young pigs were found to be positive for ETEC toxin genes; estB （38.9%）, estB/STAP （25%）, and estB/LT （13.9%） were dominant. Risk factors for ETEC-diarrhea in pigs include： leaving sick piglets in a pen with healthy piglets [odds ratio （OR） = 33.52; P 〈 0.0001]; water spillage in pen （OR = 42.87; P 〈 0.0001）; hypothermic piglets （OR = 7.29; P 〈 0.0001）; runt piglets in pen with healthy littermates （OR = 3.65; P 〈 0.0001）; and prolonged use of antibiotics （OR = 3.05; P = 0.05）.
基金This research was funded by Shaanxi Administration of Traditional Chinese Medicine Projects(2021-QYZL-01,2021-QYPT-001)Key R&D Project in Shaanxi Province(2023-YBSF-463)+1 种基金the Foundation of Science and Technology in Shaanxi Province(2020TD-050)the Fundamental Research Funds for the Central Universities(GK202205010).
文摘Background:The forest musk deer,a rare fauna species found in China,is famous for its musk secretion which is used in selected Traditional Chinese medicines.However,over-hunting has led to musk deer becoming an endangered species,and their survival is also greatly challenged by various high incidence and high mortality respiratory and intestinal diseases such as septic pneumonia and enteritis.Accumulating evidence has demonstrated that Akkermannia muciniphila(AKK)is a promising probiotic,and we wondered whether AKK could be used as a food additive in animal breeding pro-grammes to help prevent intestinal diseases.Methods:We isolated one AKK strain from musk deer feces(AKK-D)using an im-proved enrichment medium combined with real-time PCR.After confirmation by 16S rRNA gene sequencing,a series of in vitro tests was conducted to evaluate the probiotic effects of AKK-D by assessing its reproductive capability,simulated gas-trointestinal fluid tolerance,acid and bile salt resistance,self-aggregation ability,hy-drophobicity,antibiotic sensitivity,hemolysis,harmful metabolite production,biofilm formation ability,and bacterial adhesion to gastrointestinal mucosa.Results:The AKK-D strain has a probiotic function similar to that of the standard strain in humans(AKK-H).An in vivo study found that AKK-D significantly amelio-rated symptoms in the enterotoxigenic Escherichia coli(ETEC)-induced murine diar-rhea model.AKK-D improved organ damage,inhibited inflammatory responses,and improved intestinal barrier permeability.Additionally,AKK-D promoted the reconsti-tution and maintenance of the homeostasis of gut microflora,as indicated by the fact that AKK-D-treated mice showed a decrease in Bacteroidetes and an increase in the proportion of other beneficial bacteria like Muribaculaceae,Muribaculum,and unclas-sified f_Lachnospiaceae compared with the diarrhea model mice.Conclusion:Taken together,our data show that this novel AKK-D strain might be a potential probiotic for use in musk deer breeding,although further extensive system-atic research is still needed.
文摘Several technical parameters were studied during the fermentation of recombinant E.coli for the production of collagen-like biopolymer.The effects of dissolved oxygen as well as glucose concentration on fermentation were observed.The OD 600 value could reach 98 when dissolved oxygen was controlled at 50% and glucose around 1%.The production of human-like collagen with a yield of 29.4% was obtained.
