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1.
Sci Rep ; 10(1): 12922, 2020 07 31.
Article in English | MEDLINE | ID: mdl-32737335

ABSTRACT

Deficiencies in methyl-donor molecules (folate, B12 vitamin), DNA methylation alteration and high prevalence of Adherent-Invasive Escherichia coli (AIEC) are frequently observed in Crohn's disease (CD) patients. AIEC bacteria adhere to the enterocytes through abnormally expressed carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6) glycoprotein on host cells. This work aims at studying the relationship between methyl-donor molecules and AIEC-induced intestinal inflammatory response. CEABAC10 mice, a mouse model of CD, were fed a control or Methyl-donor Supplemented diet (MS diet). CEACAM6 promoter was hypermethylated in intestinal epithelial cells from mice fed an MS diet, which was associated with a significant decrease in CEACAM6 expression. Transcriptomic analysis revealed increased expression of anti-microbial peptides, increase in HSP70 gene family expression and a decreased expression of inflammatory marker Calprotectin upon MS diet, associated to a lower ability of AIEC bacteria to colonize gut mucosa. We observed in a cohort of CD patients that serum folate concentration was inversely correlated to Crohn's disease endoscopic index of severity and to fecal inflammatory markers. This study demonstrates that methyl-donor supplementation through the diet induces a specific intestinal micro-environment limiting pathobiont colonization of the gut. Clinicians may wish to consider methyl-donor supplementation for methyl-donor deficient CD patients.


Subject(s)
Antigens, CD/biosynthesis , Cell Adhesion Molecules/biosynthesis , Crohn Disease , DNA Methylation , Escherichia coli Infections , Escherichia coli/metabolism , Food, Formulated , GPI-Linked Proteins/biosynthesis , Intestinal Mucosa , Promoter Regions, Genetic , Animals , Antigens, CD/genetics , Bacterial Adhesion , Cell Adhesion Molecules/genetics , Crohn Disease/diet therapy , Crohn Disease/genetics , Crohn Disease/metabolism , Crohn Disease/microbiology , Disease Models, Animal , Escherichia coli Infections/diet therapy , Escherichia coli Infections/genetics , Escherichia coli Infections/metabolism , Escherichia coli Infections/pathology , Female , GPI-Linked Proteins/genetics , Gene Expression Regulation , Humans , Intestinal Mucosa/metabolism , Intestinal Mucosa/microbiology , Male , Mice , Mice, Transgenic
2.
BMC Vet Res ; 16(1): 245, 2020 Jul 14.
Article in English | MEDLINE | ID: mdl-32664940

ABSTRACT

BACKGROUND: Impaired gut microbiota leads to pathogenic bacteria infection, pro-inflammatory response and post-weaning diarrhea. Enterotoxigenic Escherichia coli (ETEC) K88 is a major cause of post-weaning diarrhea in weaned piglets. Fermented soybean meal (FSBM) could relieve diarrhea, alleviate inflammatory response, and modulate gut microbiota of weaned piglets. We used ETEC K88-challenged weaned piglet model to investigate the effects of FSBM on the growth performance, inflammatory response and cecal microbiota. Twenty-four crossbred piglets (6.8 ± 0.5 kg; 21 ± 2 days of age) were allotted into 2 treatment fed the diets with or without FSBM (6% at the expense of soybean meal). Six weaned piglets in each diet treatment were challenged by ETEC K88 (1 × 109 CFU/piglets) on day 15. The experimental period lasted for 20 days. RESULTS: The ETEC K88 challenge decreased (p < 0.05) fecal consistency and plasma interleukin-10 (IL-10) concentration, while increased (p < 0.05) average daily feed intake (ADFI) and plasma tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß) and interleukin 6 (IL-6) concentrations. After ETEC K88 challenge, dietary FSBM replacement increased (p < 0.05) final body weight (BW), average daily gain (ADG), ADFI, and fecal consistency, but decreased feed conversion ratio (FCR). The plasma IL-10 concentration of weaned piglets fed FSBM was higher (p < 0.05), while IL-1ß, IL-6 and TNF-α concentrations were lower (p < 0.05). Dietary FSBM replacement attenuated the increase of plasma TNF-α concentration and the decrease of ADG induced by ETEC K88 challenge (p < 0.05). High-throughput sequencing of 16S rRNA gene V4 region of cecal microbiota revealed that ETEC K88 challenge increased (p < 0.05) Campylobacter relative abundance on genus level. Dietary FSBM replacement resulted in higher (p < 0.05) relative abundances of Bacteroidetes and Prevotellaceae_NK3B31_group, and lower (p < 0.05) relative abundances of Proteobacteria and Actinobacillus. Furthermore, dietary FSBM replacement relieved the increase of Escherichia-Shigella relative abundance in weaned piglets challenged by ETEC K88 (p < 0.05). CONCLUSIONS: Dietary FSBM replacement improved growth performance and alleviated the diarrhea of weaned piglets challenged with ETEC K88, which could be due to modulation of cecal microbiota composition and down-regulation of inflammatory cytokines production.


Subject(s)
Animal Feed/analysis , Escherichia coli Infections/veterinary , Glycine max , Swine Diseases/diet therapy , Animals , Bacteria/classification , Cecum/microbiology , Cytokines/blood , Diarrhea/diet therapy , Diarrhea/microbiology , Diarrhea/veterinary , Diet/veterinary , Enterotoxigenic Escherichia coli , Escherichia coli Infections/diet therapy , Escherichia coli Infections/microbiology , Female , Fermentation , Gastrointestinal Microbiome/drug effects , Male , Sus scrofa , Swine , Swine Diseases/microbiology
3.
Int Immunopharmacol ; 78: 105798, 2020 Jan.
Article in English | MEDLINE | ID: mdl-31784403

ABSTRACT

The objective of the present study was to evaluate the effects of low-molecular-weight chitosan (LMWC) on the growth performance, immune responses and intestinal health of weaned pigs challenged by enterotoxigenic Escherichia coli (ETEC). A total of 32 weaned pigs were randomly allocated to four treatments: non-challenged (fed with basal diet), ETEC-challenged (fed with basal diet) and ETEC-challenged plus 50 or 100 mg/kg LMWC supplementation, respectively. After 11 days feeding, the non-challenged pigs were infused with sterilised Luria-Bertani culture, while the remaining pigs were infused with 2.6 × 1011 colony-forming units of ETEC. At 3 days post-challenge, all pigs were administered d-xylose at 0.1 g/kg body weight. One hour later, blood samples were obtained, and the pigs then euthanised to collect intestinal samples. Data showed that only 100 mg/kg LMWC supplementation attenuated (P < 0.05) the average daily gain reduction caused by ETEC. Furthermore, besides the decreased (P < 0.05) serum tumour necrosis factor-α and immunoglobulin (Ig) G concentrations detected in ETEC-challenged pigs supplemented with LMWC at 50 or 100 mg/kg, the higher dose (100 mg/kg) also decreased (P < 0.05) the serum IgM concentration and increased (P < 0.05) the villus height and villus height-to-crypt depth ratio in both the jejunum and ileum, and the sucrase activity in the ileal mucosa. Moreover, LMWC supplementation (50 or 100 mg/kg) in ETEC-challenged pigs elevated (P < 0.05) the mRNA levels of jejunal mucosal peptide transporter 1 and ileal mucosal peptide transporter 1, divalent metal transporter 1 and zinc transporter 1, and decreased (P < 0.05) the ileal and caecal E. coli abundances, while 100 mg/kg LMWC additionally elevated (P < 0.05) the ileal Bacillus abundance, and caecal and colonic Bifidobacterium abundances. These results suggest that LMWC helps alleviate ETEC-induced growth retardation in weaned pigs, which could be associated with the inhibition of the immune responses and improved intestinal health.


Subject(s)
Chitosan/therapeutic use , Dietary Supplements , Enterotoxigenic Escherichia coli , Escherichia coli Infections/diet therapy , Growth Disorders/diet therapy , Animals , Chitosan/chemistry , Cytokines/blood , Escherichia coli Infections/blood , Escherichia coli Infections/complications , Escherichia coli Infections/pathology , Growth Disorders/blood , Growth Disorders/etiology , Growth Disorders/pathology , Immunoglobulins/blood , Intestines/drug effects , Intestines/enzymology , Intestines/pathology , Lactase/blood , Molecular Weight , Sucrase/blood , Swine , Weaning , alpha-Glucosidases/blood
4.
Curr Protein Pept Sci ; 21(8): 772-776, 2020.
Article in English | MEDLINE | ID: mdl-31724511

ABSTRACT

Dietary proteins are linked to the pathogenic Escherichia coli (E. coli) through the intestinal tract, which is the site where both dietary proteins are metabolized and pathogenic E. coli strains play a pathogenic role. Dietary proteins are degraded by enzymes in the intestine lumen and their metabolites are transferred into enterocytes to be further metabolized. Seven diarrheagenic E. coli pathotypes have been identified, and they damage the intestinal epithelium through physical injury and effector proteins, which lead to inhibit the digestibility and absorption of dietary proteins in the intestine tract. But the increased tryptophan (Trp) content in the feed, low-protein diet or milk fractions supplementation is effective in preventing and controlling infections by pathogenic E. coli in the intestine.


Subject(s)
Diarrhea/metabolism , Dietary Proteins/metabolism , Enteropathogenic Escherichia coli/metabolism , Enterotoxigenic Escherichia coli/metabolism , Escherichia coli Infections/metabolism , Shiga-Toxigenic Escherichia coli/metabolism , Animal Feed/analysis , Animals , Diarrhea/diet therapy , Diarrhea/microbiology , Diarrhea/pathology , Diet, Protein-Restricted/methods , Dietary Proteins/therapeutic use , Enteropathogenic Escherichia coli/drug effects , Enteropathogenic Escherichia coli/pathogenicity , Enterotoxigenic Escherichia coli/drug effects , Enterotoxigenic Escherichia coli/pathogenicity , Escherichia coli Infections/diet therapy , Escherichia coli Infections/microbiology , Escherichia coli Infections/pathology , Gastrointestinal Microbiome , Humans , Intestinal Absorption/drug effects , Intestinal Mucosa/drug effects , Intestinal Mucosa/metabolism , Intestinal Mucosa/microbiology , Intestinal Mucosa/pathology , Milk Proteins/metabolism , Milk Proteins/therapeutic use , Shiga-Toxigenic Escherichia coli/drug effects , Shiga-Toxigenic Escherichia coli/pathogenicity , Tryptophan/metabolism , Tryptophan/pharmacology
5.
Microb Pathog ; 138: 103849, 2020 Jan.
Article in English | MEDLINE | ID: mdl-31704465

ABSTRACT

Avian colibacillosis is one of the most serious infectious bacterial diseases that endanger the modern poultry industry. Lactobacillus is believed to inhibit intestinal pathogens and maintain a healthy gut microbiota. This study aimed to investigate Lactobacillus supplementation in Cherry Valley ducks to prevent the intestinal flora dysbiosis caused by Duck Escherichia coli 17. One hundred and twenty healthy one day old Cherry Valley ducks were randomized to three study groups (Group I = the control group; Group II = duck Escherichia coli 17 challenge group and Group III = DE17 challenge group supplemented with lactic acid bacteria composite preparation). Cherry Valley ducks in Group II and Group III were gavage challenged with DE17 (1 × 105 CFU/mL) on day 14. Pyrosequencing of the V3/V4 variable regions of the genes encoding for 16S rRNA was used for sequence analysis. The results showed that the normal intestinal microecology was affected by DE17, including a relative increase in proteobacteria. At the same time, the Lactobacillales were increased and harmful bacteria were decreased in different intestinal segments of ducks in Group III, compared to those in Group II. Network analysis showed that dietary lactic acid bacteria addition improved the interaction pattern within the cecal microbiota of ducks and the result showed that in Ruminococcus_2 was independently present in the group III and Lachnospiraceae_NK4A136_group species correlation existed between group I and group III. This study proved that oral supplementation with Lactobacillus casei 1.2435, Lactobacillus rhamnosus 621 and Lactobacillus rhamnosus A4 can mitigate DE17 induced intestinal flora dysbiosis.


Subject(s)
Cecum/microbiology , Ducks , Escherichia coli Infections/veterinary , Escherichia coli/pathogenicity , Probiotics/pharmacology , Animals , Dietary Supplements , Ducks/microbiology , Dysbiosis , Escherichia coli Infections/diet therapy , Gastrointestinal Microbiome/genetics , Lactobacillus , Lacticaseibacillus rhamnosus , RNA, Ribosomal, 16S/genetics
6.
Am J Physiol Renal Physiol ; 316(5): F814-F822, 2019 05 01.
Article in English | MEDLINE | ID: mdl-30724105

ABSTRACT

Iron is a critical nutrient required by hosts and pathogens. Uropathogenic Escherichia coli (UPEC), the principal causative agent of urinary tract infections (UTIs), chelate iron for their survival and persistence. Here, we demonstrate that dietary modulation of iron availability limits UPEC burden in a mouse model of UTI. Mice on a low-iron diet exhibit reduced systemic and bladder mucosal iron availability and harbor significantly lower bacterial burden, concomitant with dampened inflammation. Hepcidin is a master regulator of iron that controls iron-dependent UPEC intracellular growth. Hepcidin-deficient mice ( Hamp1-/-) exhibit accumulation of iron deposits, persistent bacterial burden in the bladder, and a heightened inflammatory response to UTI. However, a low-iron dietary regimen reversed the iron overload and increased bacterial burden phenotypes in Hamp1-/- mice. Thus modulation of iron levels via diet can reduce UPEC infection and persistence, which may have significant implications for clinical management of UTI.


Subject(s)
Escherichia coli Infections/diet therapy , Iron, Dietary/metabolism , Urinary Bladder/microbiology , Urinary Tract Infections/diet therapy , Uropathogenic Escherichia coli/pathogenicity , Animals , Bacterial Load , Disease Models, Animal , Escherichia coli Infections/metabolism , Escherichia coli Infections/microbiology , Ferritins/metabolism , Hepcidins/genetics , Hepcidins/metabolism , Host-Pathogen Interactions , Interleukin-6/metabolism , Mice, Inbred C57BL , Mice, Knockout , Urinary Bladder/metabolism , Urinary Tract Infections/metabolism , Urinary Tract Infections/microbiology
7.
Infect Disord Drug Targets ; 18(3): 199-206, 2018.
Article in English | MEDLINE | ID: mdl-29621966

ABSTRACT

OBJECTIVES: There is limited published data concerning the recent epidemiology of urinary tract infections (UTI) in HIV-patients, thus we analysed independent risk factors for UTI in HIV positive individuals and antimicrobial resistance rates of E. coli to antimicrobial agents commonly used in UTI. To determine the prevalence of symptomatic urinary tract infections (UTI) in HIV-patients, we performed a retrospective case-control study. METHODS: We included 313 HIV-patients, 101 with UTI and 212 age and gendermatched controls, attending the HIV outpatient clinic at the Vienna University Hospital (VUH) over a period from January 2011 to September 2016. The patients' specific data was gathered from the electronic database of the VUH. The statistical analysis was performed using SPSS Software Version 20.0. RESULTS: HIV infected individuals with CD4 count >200 cells/mm3 were less likely than HIV infected individuals with CD4 count <200 cells/mm3 to experience UTI (OR 0.811, 95% CI 0.712-0.923 vs. OR 2.555, 95% CI 1.553 - 4.205, respectively). The in vitro resistance rate of E. coli to antimicrobial agents was as follows: ciprofloxacin (41%), mecillinam (20.5%), trimethoprim (61%), ampicillin (67%), ampicillin/ clavulanic acid (23%), cefuroxime (17%), nitrofurantoin (2%), amikacin (0%) and gentamicin (9.5%). CONCLUSION: Immunological status (CD4 count) is an important parameter for risk assessment of UTIs in HIV-patients. The increased resistance rate of E. coli to commonly used antimicrobial agents needs to be considered when it comes to the management of UTI, additionally, surveillance strategies should be implemented in HIV-patients.


Subject(s)
Escherichia coli Infections/immunology , Escherichia coli/immunology , HIV Infections/epidemiology , Urinary Tract Infections/epidemiology , Adolescent , Adult , Anti-Infective Agents/therapeutic use , CD4 Lymphocyte Count , Case-Control Studies , Drug Resistance, Multiple, Bacterial/drug effects , Escherichia coli/drug effects , Escherichia coli Infections/diet therapy , Escherichia coli Infections/epidemiology , Escherichia coli Infections/microbiology , Europe/epidemiology , Female , HIV Infections/drug therapy , HIV Infections/immunology , Hospitals, University , Humans , Male , Middle Aged , Prevalence , Retrospective Studies , Risk Factors , Tertiary Healthcare , Urinary Tract Infections/drug therapy , Urinary Tract Infections/microbiology
8.
J Nutr ; 147(11): 2050-2059, 2017 11.
Article in English | MEDLINE | ID: mdl-28954839

ABSTRACT

Background: Diarrheal diseases in infancy and childhood are responsible for substantial morbidity and mortality in developing nations. Lysozyme, an antimicrobial component of human milk, is thought to play a role in establishing a healthy intestinal microbiota and immune system. Consumption of breast milk has been shown to prevent intestinal infections and is a recommended treatment for infants with diarrhea.Objective: This study aimed to examine the ability of lysozyme-rich goat milk to prevent intestinal infection.Methods: Six-week-old Hampshire-Yorkshire pigs were assigned to treatment groups balanced for weight, sex, and litter and were fed milk from nontransgenic control goats (GM group) or human lysozyme transgenic goats (hLZM group) for 2 wk before they were challenged with porcine-specific enterotoxigenic Escherichia coli (ETEC). Fecal consistency, complete blood counts, intestinal histology, and microbial populations were evaluated.Results: Pigs in the hLZM group had less severe diarrhea than did GM pigs at 24 and 48 h after ETEC infection (P = 0.01 and 0.05, respectively), indicating a less severe clinical disease state. Relative to baseline, postmilk hLZM pigs had 19.9% and 137% enrichment in fecal Bacteroidetes (P = 0.028) and Paraprevotellaceae (P = 0.003), respectively, and a 93.8% reduction in Enterobacteriaceae (P = 0.007), whereas GM pigs had a 60.9% decrease in Lactobacillales (P = 0.003) and an 83.3% enrichment in Burkholderiales (P = 0.010). After ETEC infection, hLZM pigs tended to have lower amounts (68.7% less) of fecal Enterobacteriaceae than did GM pigs (P = 0.058). There were 83.1% fewer bacteria translocated into the mesenteric lymph nodes of hLZM pigs than into those of GM pigs (P = 0.039), and hLZM pigs had 34% lower mucin 1 and 61% higher tumor necrosis factor-α expression in the ileum than did GM pigs (P = 0.046 and 0.034, respectively).Conclusion: Results of this study indicate that human lysozyme milk consumption before and during ETEC infection has a positive effect on clinical disease, intestinal mucosa, and gut microbiota in young pigs.


Subject(s)
Escherichia coli Infections/veterinary , Intestinal Diseases/veterinary , Milk/chemistry , Muramidase/administration & dosage , Swine Diseases/diet therapy , Animal Feed/analysis , Animals , Animals, Genetically Modified , Animals, Newborn , Bacteroidetes , Diet/veterinary , Disease Models, Animal , Enterotoxigenic Escherichia coli , Escherichia coli Infections/diet therapy , Feces/microbiology , Gastrointestinal Microbiome , Goats/genetics , Intestinal Diseases/diet therapy , Intestines/microbiology , Muramidase/analysis , Swine , Swine Diseases/microbiology
9.
Sci Rep ; 7(1): 5439, 2017 07 14.
Article in English | MEDLINE | ID: mdl-28710379

ABSTRACT

The aim of this study was to investigate the effects of dietary supplementation with two alternatives to antibiotics (Candida tropicalis and mulberry leaf flavonoids) on intestinal microbiota of preweaned calves challenged with Escherichia coli K99. Sixty Holstein calves were randomly assigned to 5 treatments: fed a basal diet (N-CON); fed a basal diet and challenged with E.coli K99 (P-CON); fed a basal diet supplemented with C.tropicalis (CT), mulberry leaf flavonoids (MLF), and the combination of the two additives (CM), respectively, and challenged with E.coli K99. The MLF and CM groups had significantly higher average daily grain and feed efficiency, and significantly lower fecal scores compared with the P-CON group after E. coli K99 challenge. The supplementation groups increased the relative abundance, at the phylum level, of Bacteroidetes and Proteobacteria, whereas at the genus level, they increased the relative abundance of Prevotella, Lactobacillus, and Enterococcus. Quantitative PCR revealed that the CT, MLF, and CM groups had significantly lower copy numbers of E.coli K99 compared with the P-CON group. The CT, MLF, and CM treatments reduce days of diarrhea, improve intestinal health, and beneficially manipulate the intestinal microbiota in preweaned calves.


Subject(s)
Candida tropicalis/physiology , Cattle Diseases/diet therapy , Diarrhea/veterinary , Dietary Supplements , Escherichia coli Infections/veterinary , Flavonoids/administration & dosage , Morus/chemistry , Animal Feed/analysis , Animals , Antibiosis/genetics , Bacterial Typing Techniques , Bacteroidetes/classification , Bacteroidetes/genetics , Bacteroidetes/isolation & purification , Cattle , Cattle Diseases/microbiology , Cattle Diseases/pathology , Diarrhea/diet therapy , Diarrhea/microbiology , Diarrhea/pathology , Enterococcus/classification , Enterococcus/genetics , Enterococcus/isolation & purification , Escherichia coli/genetics , Escherichia coli/growth & development , Escherichia coli/pathogenicity , Escherichia coli Infections/diet therapy , Escherichia coli Infections/microbiology , Escherichia coli Infections/pathology , Feces/microbiology , Gastrointestinal Microbiome/genetics , Lactobacillus/classification , Lactobacillus/genetics , Lactobacillus/isolation & purification , Male , Plant Leaves/chemistry , Polymerase Chain Reaction , Prevotella/classification , Prevotella/genetics , Prevotella/isolation & purification , Proteobacteria/classification , Proteobacteria/genetics , Proteobacteria/isolation & purification , Weaning
10.
Int J Adolesc Med Health ; 29(2)2017 Apr 01.
Article in English | MEDLINE | ID: mdl-26556838

ABSTRACT

A tubo-ovarian abscess is a rare presentation in non-sexually active adolescents; only 11 cases have been reported in the literature. Variable approaches for diagnosis and management are described. We present a 19-year-old, non-sexually active, medically free girl, who had an abdominopelvic mass with abdominal pain and vomiting followed by fever. She had a confusing presentation of malignancy versus tuberculosis, with the help of imaging, diagnosis and treatment with percutaneous drainage, conservative treatment was achieved. Diagnosis of a tubo-ovarian abscess is difficult in non-sexually active adolescents, a high clinical index of suspicion is important as misdiagnosis may lead to radical and aggressive management, conservative management is possible in many of these patients.


Subject(s)
Abscess/diagnostic imaging , Escherichia coli Infections/diet therapy , Ovarian Diseases/diagnostic imaging , Sexual Abstinence , Abscess/drug therapy , Abscess/microbiology , Adult , Anti-Bacterial Agents/administration & dosage , Diagnosis, Differential , Escherichia coli/isolation & purification , Escherichia coli Infections/drug therapy , Female , Humans , Ovarian Diseases/drug therapy , Ovarian Diseases/microbiology , Piperacillin/administration & dosage , Treatment Outcome , Young Adult
11.
Dig Dis Sci ; 62(4): 922-933, 2017 04.
Article in English | MEDLINE | ID: mdl-27995406

ABSTRACT

BACKGROUND AND AIM: Enterotoxigenic Escherichia coli (ETEC) strains are involved in piglet post-weaning diarrhea. Prophylactic measures including probiotics have been examined in infection experiments with live piglets. In the present study, we have tested whether the early effects of ETEC infection can also be evoked and studied in a model in which ETEC is added to whole mucosal tissues ex vivo, and whether this response can be modulated by prior supplementation of the piglets with probiotics. METHODS: Jejunal barrier and transport properties of Enterococcus faecium-supplemented or control piglets were assessed in Ussing chambers. Part of the epithelia was challenged with an ETEC strain at the mucosal side. Fluxes of fluorescein as a marker of paracellular permeability, and the expression of selected tight junction (TJ) proteins and of proinflammatory cytokines were measured. RESULTS: The addition of ETEC ex vivo induced an increase in transepithelial resistance peaking in the first 2 h with a concomitant reduction in fluorescein fluxes, indicating tightening effects on barrier function. The response of short-circuit current after stimulation with PGE2 or glucose was reduced in epithelia treated with ETEC. ETEC induced a decrease in the TJ protein claudin-4 in the control diet group after 280 min and an increase in the mRNA expression of the proinflammatory cytokines interleukin-8 and TNF-α in both groups after 180 min. CONCLUSIONS: The addition of ETEC ex vivo affected barrier function and transport properties of the jejunal tissues and enhanced cytokine expression. The differences in claudin-4 expression in the jejunum might indicate a beneficial effect of E. faecium prefeeding.


Subject(s)
Cytokines/biosynthesis , Enterotoxigenic Escherichia coli , Intestinal Mucosa/metabolism , Intestinal Mucosa/microbiology , Jejunum/metabolism , Jejunum/microbiology , Probiotics/administration & dosage , Animal Feed , Animals , Cytokines/genetics , Escherichia coli Infections/diet therapy , Escherichia coli Infections/metabolism , Escherichia coli Infections/prevention & control , Female , Gene Expression , Intestinal Mucosa/drug effects , Jejunum/drug effects , Male , Pregnancy , Random Allocation , Swine
12.
PLoS One ; 11(8): e0160994, 2016.
Article in English | MEDLINE | ID: mdl-27575007

ABSTRACT

Sutherlandia frutescens is a medicinal plant that has been traditionally used in southern Africa for cancers, infections, and inflammatory conditions. We recently published experiments demonstrating that an aqueous extract of S. frutescens possessed potent immune-stimulatory activity. This work was carried out with murine macrophages, an immune cell type that plays a pivotal role in host defense from infection and in shaping host inflammatory and immune responses. Here, we conducted a series of follow-up experiments to explore the impact of consuming S. frutescens on host response to bacterial challenge using healthy mice. We found that feeding mice a diet containing S. frutescens failed to significantly alter host response to systemic infection by either a gram-positive or gram-negative bacterium (i.e., L. monocytogenes and E. coli, respectively). In contrast to the in vitro observations, we found no evidence that S. frutescens consumption stimulated in vivo inflammatory responses; instead, consumption of S. frutescens tended to diminish in vivo inflammatory responses. Several possible reasons for this are discussed.


Subject(s)
Cytokines/metabolism , Escherichia coli Infections/immunology , Fabaceae/chemistry , Listeriosis/immunology , Plant Extracts/administration & dosage , Administration, Oral , Africa, Southern , Animals , Cells, Cultured , Escherichia coli/drug effects , Escherichia coli Infections/diet therapy , Female , Gene Expression Regulation/drug effects , Listeria monocytogenes/drug effects , Listeriosis/diet therapy , Macrophages/cytology , Macrophages/drug effects , Macrophages/immunology , Male , Mice , Plant Extracts/pharmacology , Plants, Medicinal/chemistry
14.
Mol Nutr Food Res ; 59(7): 1292-306, 2015 Jul.
Article in English | MEDLINE | ID: mdl-25917127

ABSTRACT

Urinary tract infections (UTI) are one of the most frequent extraintestinal infections caused by Escherichia coli (ExPEC). Cranberry juice has been used for decades to alleviate symptoms and prevent recurrent UTI. The putative compounds in cranberries are proanthocyanidins (PAC), specifically PAC with "A-type" bonds. Since PAC are not absorbed, their health benefits in UTI may occur through interactions at the mucosal surface in the gastrointestinal tract. Recent research showed that higher agglutination of ExPEC and reduced bacterial invasion are correlated with higher number of "A-type" bonds and higher degree of polymerization of PAC. An understanding of PAC structure-activity relationship is becoming feasible due to advancements, not only in obtaining purified PAC fractions that allow accurate estimation, but also in high-resolution MS methodologies, specifically, MALDI-TOF MS. A recent MALDI-TOF MS deconvolution method allows quantification of the ratios of "A-type" to "B-type" bonds enabling characteristic fingerprints. Moreover, the generation of fluorescently labeled PAC allows visualization of the interaction between ExPEC and PAC with microscopy. These tools can be used to establish structure-activity relationships between PAC and UTI and give insight on the mechanism of action of these compounds in the gut without being absorbed.


Subject(s)
Proanthocyanidins/chemistry , Proanthocyanidins/pharmacology , Uropathogenic Escherichia coli/drug effects , Vaccinium macrocarpon/chemistry , Biological Availability , Chromatography/methods , Escherichia coli Infections/diet therapy , Humans , Proanthocyanidins/pharmacokinetics , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Spectrophotometry/methods , Structure-Activity Relationship , Urinary Tract Infections/diet therapy , Urinary Tract Infections/microbiology , Uropathogenic Escherichia coli/pathogenicity
15.
J Appl Microbiol ; 117(1): 217-26, 2014 Jul.
Article in English | MEDLINE | ID: mdl-24674595

ABSTRACT

AIMS: To identify a fast, economic and reliable method for preselecting lactic acid-producing bacterial (LAB) isolates to control enterotoxigenic Escherichia coli (ETEC). METHODS AND RESULTS: Two assays with porcine intestinal epithelial IPEC-J2 cells or Caenorhabditis elegans for selecting effective probiotic candidates were compared. Both assays were based on measuring death of cells or worms caused by ETEC strain JG280. Six of 13 LAB isolates showed ≥50% protection in each assay, among which only four isolates (≥50% protection) were consistently selected by both assays. Isolate CL9 (Lactobacillus reuteri) was further studied. It reduced gene expression of estA, estB and elt in JG280 in both assays. Furthermore, the isolate protected IPEC-J2 and C. elegans from cell and worm death caused by STa, STb or LT enterotoxin expressed in E. coli DH5α. CL9 also promoted host defensive responses by decreasing IL-8 and increasing IL-10 production in IPEC-J2 cells and expression of antimicrobial peptide genes clec-60, clec-85 in C. elegans. CONCLUSIONS: Caenorhabditis elegans is useful for preselecting probiotic candidates to control ETEC after initial screening with IPEC-J2 cells. SIGNIFICANCE AND IMPACT OF THE STUDY: A combination of IPEC-J2 cell and C. elegans assays can improve the effectiveness in preselecting probiotic candidates.


Subject(s)
Caenorhabditis elegans/drug effects , Enterotoxigenic Escherichia coli/drug effects , Epithelial Cells/drug effects , Limosilactobacillus reuteri/physiology , Probiotics/pharmacology , Animals , Antibiosis , Antimicrobial Cationic Peptides/agonists , Antimicrobial Cationic Peptides/biosynthesis , Caenorhabditis elegans/microbiology , Cell Line , Enterotoxigenic Escherichia coli/growth & development , Enterotoxins/antagonists & inhibitors , Enterotoxins/biosynthesis , Epithelial Cells/microbiology , Escherichia coli Infections/diet therapy , Escherichia coli Infections/microbiology , Escherichia coli Infections/veterinary , Interleukin-10/agonists , Interleukin-10/metabolism , Interleukin-8/antagonists & inhibitors , Interleukin-8/metabolism , Intestines/drug effects , Intestines/microbiology , Models, Biological , Swine , Swine Diseases/diet therapy , Swine Diseases/microbiology
16.
PLoS One ; 8(6): e66280, 2013.
Article in English | MEDLINE | ID: mdl-23840434

ABSTRACT

N-carbamylglutamate (NCG) has been shown to enhance performance in neonatal piglets. However, few studies have demonstrated the effect of NCG on the intestinal mucosal barrier. This study was conducted to determine the effects of dietary NCG supplementation on intestinal mucosal immunity in neonatal piglets after an Escherichia coli (E. coli) challenge. New-born piglets (4 d old) were assigned randomly to one of four treatments (n = 7), including (I) sham challenge, (II) sham challenge +50 mg/kg NCG, (III) E. coli challenge, and (IV) E. coli challenge +50 mg/kg NCG. On d 8, pigs in the E. coli challenge groups (III and IV) were orally challenged with 5 mL of E. coli K88 (10(8) CFU/mL), whereas pigs in the sham challenge groups (I and II) were orally dosed with an equal volume of water. On d 13, all piglets were sacrificed, and samples were collected and examined. The results show that average daily gain in the E. coli challenged piglets (III and IV) was decreased (PE.coli<0.05). However, it tended to be higher in the NCG treated piglets (II and IV). Ileum secretory IgA, as well as IFN-γ, IL-2, IL-4 and IL-10 in ileal homogenates, were increased in E. coli challenged piglets (III and IV). Similarly, ileum SIgA and IL-10 levels, and CD4(+) percentage in NCG treated piglets (II and IV) were higher than no-NCG treated piglets (PNCG<0.05). However, the IL-2 level was only decreased in the piglets of E. coli challenge + NCG group (IV) compared with E. coli challenge group (III) (P<0.05). No change in the IL-2 level of the sham challenged piglets (III) was observed. In conclusion, dietary NCG supplementation has some beneficial effects on intestinal mucosal immunity in E. coli challenged piglets, which might be associated with stimulated lymphocyte proliferation and cytokine synthesis. Our findings have an important implication that NCG may be used to reduce diarrhea in neonatal piglets.


Subject(s)
Cytokines/metabolism , Escherichia coli Infections/diet therapy , Glutamates/administration & dosage , Intestinal Mucosa/immunology , Swine Diseases/diet therapy , Animals , Animals, Newborn , Dietary Supplements , Escherichia coli Infections/immunology , Escherichia coli Infections/veterinary , Glutamates/pharmacology , Immunity, Mucosal/drug effects , Intestinal Mucosa/drug effects , Male , Random Allocation , Swine , Swine Diseases/immunology , Swine Diseases/microbiology
17.
Z Gastroenterol ; 50(2): 209-12, 2012 Feb.
Article in German | MEDLINE | ID: mdl-22298100

ABSTRACT

A 29-year-old man presented with abdominal cramps and bloody diarrhoea. Blood tests revealed elevated C-reactive protein (21.3 mg/dL; normal range 0.01 - 0. 82 mg/dL) and white blood cells (28200/µL, normal range 4000 - 10000/µL). Stool tests were negative for enteropathogenic bacteria and Clostridium difficile toxins A/B. Ultrasound and computed tomography showed massive swelling of the transverse colon and right colonic flexure. At endoscopy, circular necrosis of the mucosa was encountered in the proximal segments of the colon whereas distal parts of the organ showed patchy inflammation of minor severity. Extended stool testing identified Escherichia coli type O104:H4 as the causative microorganism. There was no evidence for haemolytic uraemic syndrome. Under conservative treatment the patient recovered clinically, serologically and endoscopically. At follow-up endoscopy, longitudinal ulcers and vital mucosa were present. In this case report the segmental pattern of mucosal necrosis in a patient with EHEC infection is noteworthy.


Subject(s)
Colitis/diagnosis , Colitis/microbiology , Colon/diagnostic imaging , Colon/pathology , Enterohemorrhagic Escherichia coli/isolation & purification , Escherichia coli Infections/diet therapy , Escherichia coli Infections/microbiology , Adult , Colitis/therapy , Escherichia coli Infections/therapy , Humans , Male , Necrosis/diagnosis , Radiography
18.
Phytomedicine ; 19(6): 506-14, 2012 Apr 15.
Article in English | MEDLINE | ID: mdl-22306419

ABSTRACT

Consumption of cranberries is known to exert positive health effects, especially against urinary tract infections. For this reason, presumably, they are widely used in folk medicine. Different aspects of cranberry phenolics activity were studied in individual papers but complex study in this matter is missing. The aim of the present study is to provide complex data concerning various aspects of cranberry extract activity. We studied the effects of subinhibitory concentrations of commercially available extract (Zuravit S·O·S(®)) against two Escherichia coli strains isolated from urine of patients with pyelonephritis. Additionally the main extract anthocyanins were characterized. The activity of extract against lipid peroxidation and its radical scavenging ability were also assessed. Zuravit S·O·S(®) decreased the hydrophobicity of one of the studied E. coli strains, reduced swimming motility and adhesion to epithelial cells of both studied strains, it also limited the ability of bacteria to form biofilm. Expression of curli was not affected by cranberry extract, the assessment of P fimbriae expression was not reliable due to extract-induced agglutination of erythrocytes. Cranberry extract caused filamentation in both studied E. coli strains. It also showed pronounced antioxidant and radical scavenging properties. The properties of the studied cranberry extract show that it could be effectively used in prevention and/or elimination of urinary tract infections, specially the recurrent ones.


Subject(s)
Biofilms/drug effects , Escherichia coli Infections/diet therapy , Phytotherapy , Plant Extracts/pharmacology , Uropathogenic Escherichia coli/drug effects , Uropathogenic Escherichia coli/pathogenicity , Vaccinium macrocarpon/chemistry , Anthocyanins/pharmacology , Antioxidants/pharmacology , Drug Evaluation , Fruit/chemistry , Humans , Pyelonephritis/drug therapy , Urinary Tract Infections/diet therapy , Urinary Tract Infections/prevention & control , Urine/microbiology , Uropathogenic Escherichia coli/physiology
19.
Foodborne Pathog Dis ; 7(10): 1159-64, 2010 Oct.
Article in English | MEDLINE | ID: mdl-20590426

ABSTRACT

Enterotoxigenic Escherichia coli (ETEC) is a major cause of diarrhea among infants and children in developing countries, as well as among travelers to these areas. The major virulence factors of ETEC are the colonization factor antigens (CFAs) and a heat-labile enterotoxin (LT) and/or a heat-stable enterotoxin (ST). Among Israeli recruits serving under military field conditions, 107 of all examined isolates expressed LT or ST, and CFAs could be characterized in 68% of the isolates, in which CFAs of the CFA/II group and CS6 were the most prevalent. Additionally, 31% of the 107 ETEC isolates showed resistance to three or more of the antimicrobial agents examined, and the percentage of resistant isolates expressing LT was significantly higher than those expressing ST or LT+ST. These results may be important for development of an effective vaccine and for facilitation of an empirical choice of antibiotic treatment or prophylaxis for traveler's diarrhea in this area.


Subject(s)
Diarrhea/microbiology , Enterotoxigenic Escherichia coli/drug effects , Enterotoxigenic Escherichia coli/pathogenicity , Escherichia coli Infections/microbiology , Adolescent , Antigens, Bacterial/analysis , Bacterial Vaccines , Drug Resistance, Bacterial , Enterotoxigenic Escherichia coli/chemistry , Enterotoxins/analysis , Escherichia coli Infections/diet therapy , Escherichia coli Infections/prevention & control , Feces/microbiology , Fimbriae Proteins/analysis , Hot Temperature , Humans , Israel , Military Personnel , Phenotype , Young Adult
20.
J Anim Sci ; 87(1): 148-56, 2009 Jan.
Article in English | MEDLINE | ID: mdl-18791156

ABSTRACT

We tested the effect of Trp addition to a standard weaning diet and oral challenge with enterotoxigenic Escherichia coli K88 (ETEC) on growth and health of piglets susceptible or nonsusceptible to the intestinal adhesion of ETEC. Sixty-four pigs weaned at 21 d of age were divided into 3 groups based on their ancestry and BW: a control group of 8 pigs fed a basal diet (B), the first challenged group of 28 pigs fed B diet (BCh), and the second challenged group of 28 pigs fed a diet with Trp (TrpCh). The Trp diet was produced by the addition of 1 g of l-Trp/kg to the basal diet. On d 5, pigs were orally challenged with 1.5 mL suspension containing 10(10) cfu ETEC/mL or placebo, and killed on d 9 or 23. Based on in vitro villus adhesion assay, the pigs (except the B group) were classified as susceptible (s(+)) or nonsusceptible (s(-)) to the intestinal ETEC adhesion. Thus, after the challenge, treatments were B, BChs(-), BChs(+), TrpChs(-), and TrpChs(+). Pigs susceptible to ETEC were 50.0% in the BChs(+) group (3 pigs lost included) and 46.4% in the TrpChs (+) group (1 pig lost included). During the first 4 d after challenge, the challenge reduced ADG (P < 0.05), and this reduction was greater in susceptible pigs (P < 0.05) than nonsusceptible ones. Tryptophan increased ADG and feed intake in susceptible pigs (P < 0.05) from challenge to d 4, but not thereafter. Tryptophan supplementation did not improve the fecal consistency and did not reduce the number of pigs positive for ETEC in feces on d 4 after the challenge. The K88-specific immunoglobulin A activity in blood serum tended to be greater in challenged pigs (P = 0.102) and was not affected by the addition of Trp. Villous height was affected by the addition of Trp and challenge in different ways, depending on the site of small intestine. The need to consider the phenotype for the adhesion of the ETEC in studies with different supply of Trp was clearly evident. When compared with practical weaning standard diets, Trp supplementation allowed susceptible pigs to partially compensate for the effects of ETEC challenge by increasing feed intake and maintaining an adequate BW growth. This is of practical importance for the formulation of diets for pigs selected for lean growth because of the presence of an association between this trait and the susceptibility to the intestinal adhesion of ETEC.


Subject(s)
Diet/veterinary , Dietary Supplements , Disease Susceptibility/veterinary , Eating/physiology , Escherichia coli Infections/veterinary , Swine/physiology , Weaning , Animals , Antibodies, Bacterial/blood , Bacterial Adhesion , Disease Susceptibility/diet therapy , Escherichia coli/physiology , Escherichia coli Infections/diet therapy , Feces/microbiology , Immunoglobulin A/blood , Intestines/anatomy & histology , Intestines/microbiology , Swine/growth & development , Swine/immunology , Tryptophan/administration & dosage , Weight Gain/physiology
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