Impact of diet and synbiotics on selected gut bacteria and intestinal permeability in individuals with excess body weight - A Prospective, Randomized Study.

Overweight and obese individuals may have leaky intestinal barrier and microbiome dysbiosis. The aim of this study was to determine whether body mass reduction with diet and synbiotics in an adult person with excess body mass has an influence on the gut microbiota and zonulin concentration. The study was a single blinded trial. 60 persons with excess body mass were examined. Based on randomization, patients were qualified either to the intervention group (Synbiotic group) or to the control group (Placebo group). Anthropometric measurements, microbiological assessment of faecal samples and zonulin concentration in the stool were performed before and after observation. After 3-months, an increase in the variety of intestinal bacteria (increase in the Shannon-Weaver index and the Simpson index) and a decrease in concentration of zonulin in faecal samples were observed in the Synbiotic group. Also, statistically significant correlation between zonulin and Bifidobacterium spp. (Spearman test, R=-0.51; p=0.0040) was noticed. There were no significant relationships between the body mass, BMI and changes in the intestinal microbiota or zonulin concentrations. The use of diet and synbiotics improved the condition of the microbiota and intestinal barrier in patients in the Synbiotic group.

Proteus spp. as Putative Gastrointestinal Pathogens.

Proteus species, members of the Enterobacteriaceae family, are usually considered commensals in the gut and are most commonly recognized clinically as a cause of urinary tract infections. However, the recent identification of Proteus spp. as potential pathogens in Crohn's disease recurrence after intestinal resection serves as a stimulus to examine their potential role as gut pathogens. Proteus species possess many virulence factors potentially relevant to gastrointestinal pathogenicity, including motility; adherence; the production of urease, hemolysins, and IgA proteases; and the ability to acquire antibiotic resistance. Gastrointestinal conditions that have been linked to Proteus include gastroenteritis (spontaneous and foodborne), nosocomial infections, appendicitis, colonization of devices such as nasogastric tubes, and Crohn's disease. The association of Proteus species with Crohn's disease was particularly strong. Proteus species are low-abundance commensals of the human gut that harbor significant pathogenic potential; further investigation is needed.

The association between ethnicity and vaginal microbiota composition in Amsterdam, the Netherlands.

OBJECTIVE: To evaluate whether ethnicity is independently associated with vaginal microbiota (VMB) composition in women living in Amsterdam, the Netherlands, as has been shown for American women. METHODS: Women (18-34 years, non-pregnant, N = 610) representing the six largest ethnic groups (Dutch, African Surinamese, South-Asian Surinamese, Turkish, Moroccan, and Ghanaian) were sampled from the population-based HELIUS study. Sampling was performed irrespective of health status or healthcare seeking behavior. DNA was extracted from self-sampled vaginal swabs and sequenced by Illumina MiSeq (16S rRNA gene V3-V4 region). RESULTS: The overall prevalence of VMBs not dominated by lactobacilli was 38.5%: 32.2% had a VMB resembling bacterial vaginosis and another 6.2% had a VMB dominated by Bifidobacteriaceae (not including Gardnerella vaginalis), Corynebacterium, or pathobionts (streptococci, staphylococci, Proteus or Enterobacteriaceae). The most prevalent VMB in ethnically Dutch women was a Lactobacillus crispatus-dominated VMB, in African Surinamese and Ghanaian women a polybacterial G. vaginalis-containing VMB, and in the other ethnic groups a L. iners-dominated VMB. After adjustment for sociodemographic, behavioral and clinical factors, African Surinamese ethnicity (adjusted odds ratio (aOR) 5.1, 95% confidence interval (CI) 2.1-12.0) and Ghanaian ethnicity (aOR 4.8, 95% CI 1.8-12.6) were associated with having a polybacterial G. vaginalis-containing VMB, and African Surinamese ethnicity with a L. iners-dominated VMB (aOR 2.8, 95% CI 1.2-6.2). Shorter steady relationship duration, inconsistent condom use with casual partners, and not using hormonal contraception were also associated with having a polybacterial G. vaginalis-containing VMB, but human papillomavirus infection was not. Other sexually transmitted infections were uncommon. CONCLUSIONS: The overall prevalence of having a VMB not dominated by lactobacilli in this population-based cohort of women aged 18-34 years in Amsterdam was high (38.5%), and women of sub-Saharan African descent were significantly more likely to have a polybacterial G. vaginalis-containing VMB than Dutch women independent of modifiable behaviors.

In vitro activity of three commercial bacteriophage cocktails against multidrug-resistant Escherichia coli and Proteus spp. strains of human and non-human origin.

OBJECTIVES: Bacteriophages may represent a therapeutic alternative to treat infections caused by multidrug-resistant (MDR) pathogens. However, studies analysing their activity against MDR Enterobacteriaceae are limited. METHODS: The in vitro lytic activity of three commercial bacteriophage cocktails (PYO, INTESTI and Septaphage) was evaluated against 70 Escherichia coli and 31 Proteus spp. of human and non-human origin. Isolates were characterised by phenotypic and genotypic methods and included 82 MDR strains [44 extended-spectrum-ß-lactamase (ESBL)-producers (18 CTX-M-15-like, including ST131/ST648 E. coli); 27 plasmid-mediated AmpC ß-lactamase (pAmpC)-producers (23 CMY-2-like, including ST131 E. coli); 3 ESBL+pAmpC-producers; and 8 carbapenemase-producers]. Phage susceptibility was determined by the spot test. RESULTS: E. coli susceptibility to PYO, INTESTI and Septaphage was 61%, 67% and 9%, whereas that of Proteus spp. was 29%, 39% and 19%, respectively. For the subgroup of ESBL-producing E. coli/Proteus spp., the following susceptibility rates were recorded: PYO, 57%; INTESTI, 59%; and Septaphage, 11%. With regard to pAmpC-producers, 59%, 70% and 11% were susceptible to PYO, INTESTI and Septaphage, respectively. Five of eight carbapenemase-producers and three of four colistin-resistant E. coli were susceptible to PYO and INTESTI. CONCLUSIONS: This is the first study analysing the activity of the above three cocktails against well-characterised MDR E. coli and Proteus spp. The overall narrow spectrum of activity observed could be related to the absence of specific bacteriophages targeting these contemporary MDR strains that are spreading in different settings. Therefore, bacteriophages targeting emerging MDR pathogens need to be isolated and integrated in such biopreparations.

Antibiotic Sensitivity pattern of Bacterial Isolates of Neonatal Septicemia in Peshawar, Pakistan.

BACKGROUND: Septicemia plays an important role in neonatal morbidity and mortality, especially in developing countries. OBJECTIVE: To investigate the bacterial pathogens causing neonatal sepsis and their antibiotic susceptibility profile. METHODOLOGY: A total of 2,685 neonates aged 0-28 days were included in the study. Blood from each neonate was cultured and isolates were identified using standard biochemical tests. Antibiotic sensitivity pattern was analyzed using modified Kirby-Bauer disc diffusion method. RESULTS: Blood culture positivity was observed in 1,534 (57.1%) samples. Most of the cases (1089 counts - 71%) were of early onset sepsis while 445 (29%) were of late onset sepsis. The incidence of sepsis was higher in males 856 (55.8%) than females 678 (44.2%) with a 1:2 ratio. Similarly, 58.3% of septicemic patients were neonates with low birth weights. Twelve hundred and six (78.6%) isolates were gram negative while 328 (23.4%) were gram positive bacteria. E. coli was the dominant pathogen seen in 811 (52.8%) followed by Staphylococcus aureus 300 (19.5%), Pseudomonas 199 (13%), Klebsiella 102 (6.7%), Proteus 87 (5.7%), Staphylococcus epidermidis 28(1.8%) and Salmonella in 7 (0.5%) samples. All bacterial isolates showed high sensitivity to Imipenem, Enoxacin, Ofloxacin and Ciprofloxacin while low sensitivity was observed for other antibiotics (n = 16). The Proteus species showed high level of multiple resistances to all antibiotics (5.9%). CONCLUSION: Imipenem, Enoxacin, Ofloxacin and Ciprofloxacin can be used as an effective antibiotic regimen for treatment of bacterial sepsis in neonates.

Sonication: a valuable technique for diagnosis and treatment of periprosthetic joint infections.

BACKGROUND: Periprosthetic joint infection (PJI) is the most severe complication, following joint arthroplasty. Identification of the causal microbial factor is of paramount importance for the successful treatment. PURPOSE: The aim of this study is to compare the sonication fluid cultures derived from joint prosthetic components with the respective periprosthetic tissue cultures. METHODS: Explanted prosthesis components for suspected infection were placed into a tank containing sterile Ringer's solution and sonicated for 1 minute at 40 kHz. Sonication fluid cultures were examined for 10 days, and the number and identity of any colony morphology was recorded. In addition, periprosthetic tissue specimens (>5) were collected and cultured according to standard practice. The duration of antimicrobial interruption interval before culture sampling was recorded. RESULTS: Thirty-four patients composed the study group. Sonication fluid cultures were positive in 24 patients (70.5%). Sixteen of thirty four periprosthetic tissue cultures (47.1%) were considered positive, all revealing the same microbial species with the respective sonication fluid cultures: 3 tissue samples showed polymicrobial infection. All tissue cultures were also found positive by the sonication fluid culture. CONCLUSIONS: Sonication fluid cultures represent a cheap, easy, accurate, and sensitive diagnostic modality demonstrating increased sensitivity compared to periprosthetic tissue cultures (70.5 versus 47.1%).

Deciphering simultaneous bioelectricity generation and dye decolorization using Proteus hauseri.

This first-attempt study disclosed how and why electron-shuttling mediators were capable to stimulate bioelectricity-generating capabilities of dye-bearing microbial fuel cells (MFCs) using Proteus hauseri. Due to significant biotoxicity of 4-aminophenol (4AP) and the absence of electron-mediating potential of 3AP, only 2AP among all isomers could work as an exogenous mediator to stimulate bioelectricity generation of P. hauseri. Dye toxicity to cells on anodic biofilm in MFCs apparently affected the performance of simultaneous bioelectricity production and color removal (SBP&CR) in MFCs. Plus, dose-response analysis upon toxicity potency of reactive blue 160 revealed that cells on anodic biofilm in MFCs had a higher tolerance to reactive blue 160 than suspended cells. Apparently, augmentation of electron mediator(s) with low toxicity was a feasible means to facilitate bioelectricity-generating capability of SBP&CR.

Hemoperitoneum increases the incidence of bacterial translocation.

BACKGROUND/AIMS: Severe hemorrhage may induce bacterial translocation (BT) from the bowel. Presence of hemoperitoneum is supposed to further increase the incidence of BT. METHODOLOGY: Blood was drawn from the femoral artery of rats and hemoperitoneum was created by replacing the drawn blood. Rats were randomly segregated into 5 groups. Control group rats received a sham operation. Rats in groups 1 and 2 received mild hemorrhage (15mL blood/kg body weight withdrawn) with and without hemoperitoneum respectively. Rats in groups 3 and 4 received severe hemorrhage (25mL blood/kg body withdrawn) with and without hemoperitoneum respectively. Twenty-four hours after the above manipulation, mesentery lymph nodes, livers, spleens, and finally cecums were removed for bacterial cultures. RESULTS: Rats that received severe hemorrhage had a significantly higher incidence of BT, both in tissues and in individuals, than rats that received mild hemorrhage did. group 1 rats had a higher incidence of BT in tissues compared with group 2, although the difference in individuals was not significant. On the other hand, group 3 had a higher incidence of BT either in tissues or in individuals compared with group 4. Cecal populations of bacteria assessment showed that groups with hemoperitoneum had higher levels of bacteria in comparison with groups without hemoperitoneum. CONCLUSIONS: Severe hemorrhage in rats increases the incidence of BT and the incidence is even greater in the presence of hemoperitoneum.

[Proteus bacilli: features and virulence factors]. / Bakterie z rodzaju Proteus--cechy i czynniki chorobotwórczosci.

In this article, different aspects of virulence factors of Proteus bacilii (P. mirabilis, P. vulgaris, P. penneri i P. hauseri) are presented. These are opportunistic pathogens that cause different kinds of infections, most frequently of the urinary tract. These bacteria have developed several virulence factors, such as adherence due to the presence of fimbriae or afimbrial adhesins, invasiveness, swarming phenomenon, hemolytic activity, urea hydrolysis, proteolysis, and endotoxicity. Below we focus on data concerning the molecular basis of the pathogenicity of Proteus bacilli.

A clinical assessment of the performance of a sensor to detect crystalline biofilm formation on indwelling bladder catheters.

OBJECTIVES: To test the ability of a sensor developed to signal infection by the organisms that generate the crystalline biofilms that encrust catheters, to give an early warning that encrustation was occurring on patients' catheters, as the care of many patients undergoing long-term bladder catheterization is complicated by the encrustation and blockage of their catheters. PATIENTS AND METHODS: Twenty patients were followed prospectively for the lifetime of one of their catheters. Sensors based on cellulose acetate/bromothymol blue were placed in the urine-collection bags, which were changed as usual at weekly intervals. The bacteriology was assessed and pH determined weekly on urine samples. Photographic records were made of the sensors twice weekly. On removal, each catheter was examined for encrustation and blockage. RESULTS: Proteus mirabilis was not isolated from five patients and in these cases the sensor colour remained golden-yellow to brown. The catheters drained for the scheduled period and showed no signs of encrustation. By contrast, the sensors turned dark blue/black in the urine of all 15 patients infected with P. mirabilis. All these patients' catheters were encrusted and in 12 the catheters blocked. The mean interval between the sensor signalling and the catheter blocking was 12 days. CONCLUSION: The cellulose acetate/bromothymol blue sensors placed in the urine collection bags are capable of signalling infection by P. mirabilis. They also signal the early stages of catheter encrustation and allow catheter replacement in ample time to avoid the clinical crises and emergency referrals caused by catheter blockage.

Quantitative bacterial micro-assay for rapid detection of serum phenylalanine on dry blood-spots: application in phenylketonuria screening.

Phenylketonuria is an inherited metabolic disease, which is characterized by an increased level of serum phenylalanine. Quantitative measurement of phenylalanine in the serum of phenylketonuria patients is necessary to confirm the disease, and to distinguish phenylketonuria from other forms of hyperphenylalaninemia. In this study, we report a rapid and inexpensive micro-assay for simultaneous detection and quantitative measurement of serum phenylalanine on dry blood-spots. Analysis of the standard curve showed a broad linear range for phenylalanine from 120 to 1800 micromol/L. Application of this method, the standard Guthrie bacterial inhibition assay and a high-performance liquid chromatography (HPLC) method for analysis of 34 samples from phenylketonuria patients and control samples produced comparable results, with the regression equation Y = 0.994X + 0.996. The advantage of this method over the Guthrie bacterial inhibition assay is its ability to measure serum phenylalanine quantitatively without false positive results. The method was successfully applied to dried blood-spots, serum and whole blood. The cost per sample is approximately 20-50 US cents, which is much less than for HPLC and commercial enzyme kits. The method can be automated, and is thus suitable for neonatal and mass screening for phenylketonuria, especially in developing countries where funding is a limiting factor.

[The effect of selective bowel decontamination and mechanical bowel preparation on bacterial translocation due to intraabdominal hypertension]. / Kariniçi hipertansiyon modelinde gelisen bakteriyel translokasyon üzerine selektif bagirsak dekontaminasyon ve mekanik bagirsak temizliginin etkileri.

BACKGROUND: The objective of our study is to evaluate the preventive effects of selective digestive decontamination (SDD) and mechanical bowel preparation in rats with experimentally induced bacterial translocation. METHODS: Fourty adult male Sprague Dowley rats weighing 250-300 g. were divided equally into four groups as Group 1 (sham [control]), Group 2 (experimentally induced IAH at 19 mmHg), Group 3 ( SDD group) and Group 4 (SDD and mechanical bowel preparation with 19 mmHg intraabdominal pressure). Group 3 and 4 were treated at 12 hours intervals with oral gentamycine 5 mg/kg and IM sefotaxime 100mg/kg Mechanical bowel preparation was performed by oral administration of sodium phosphate. After 24 hours all rats were sacrified; mesenteric lymph nodes, spleen and liver biopsy specimens were harvested aseptically. Specimens were diluted and cultured in McConkey medium and the colony-forming units (CFU/gr ) were calculated. RESULTS: In Kruskal Wallis tests there were no significant differences between Group 1 and 3 or 4, and also Group 3 and 4 (p>0.05, p=0.872 respectively), while differences between Group 1 and 2, and also Group 3 and 4 were statistically significant (p<0.001) with respect to CFU/g estimates. CONCLUSION: These data indicate that selective intestinal decontamination and mechanical bowel preparation prevent bacterial translocation due to intraabdominal hypertension.

Classification of Proteus vulgaris biogroup 3 with recognition of Proteus hauseri sp. nov., nom. rev. and unnamed Proteus genomospecies 4, 5 and 6.

Strains traditionally identified as Proteus vulgaris formed three biogroups. Biogroup 1, characterized by negative reactions for indole production, salicin fermentation and aesculin hydrolysis, is now known as Proteus penneri. Biogroup 2, characterized by positive reactions for indole, salicin and aesculin, was shown by DNA hybridization (hydroxyapatite method) to be a genetic species separate from biogroup 1 and from biogroup 3 which is positive for indole production and negative for salicin and aesculin. In this study, 52 strains were examined, of which 36 strains were Proteus vulgaris biogroup 3, which included the current type strain of the species P. vulgaris (ATCC 29905T), and compared to seven strains of Proteus vulgaris biogroup 2 and nine type strains of other species in the genera Proteus, Providencia and Morganella. By DNA hybridization, these 36 strains were separated into four distinct groups, designated as Proteus genomospecies 3, 4, 5 and 6. DNAs within each separate Proteus genomospecies were 74-99% related to each other in 60 degrees C hybridization reactions with < or = 4.5% divergence between related sequences. Proteus genomospecies 3 contained the former P. vulgaris type strain and one other strain and was negative in reactions for salicin fermentation, aesculin hydrolysis and deoxyribonuclease, unlike the reactions associated with strains considered as typical P. vulgaris which are positive in reactions for salicin, aesculin and DNase. Genomospecies 3 can be distinguished from Proteus genomospecies 4, 5 and 6 because it is negative for Jordan's tartrate. Proteus genomospecies 4, containing five strains, was differentiated from Proteus penneri, genomospecies 3 and 6 and most, but not all, strains of genomospecies 5, by its ability to ferment L-rhamnose. Proteus genomospecies 5 and 6, containing 18 and 11 strains, respectively, could not be separated from each other by traditional biochemical tests, by carbon source utilization tests or SDS-PAGE of whole-cell proteins. In an earlier publication, a request was made to the Judicial Commission that the former type strain of P. vulgaris (ATCC 13315) be replaced by P. vulgaris biogroup 2 strain ATCC 29905T, a strain considered more biochemically typical of P. vulgaris strains. This would have the effect of assigning the name P. vulgaris to P. vulgaris biogroup 2. Since this request has been acceded to, the name Proteus hauseri is herein proposed for Proteus vulgaris genomospecies 3. Its type strain is ATCC 700826T. Proteus genomospecies 4, 5 and 6 will remain unnamed until better phenotypic differentiation can be accomplished. All Proteus genomospecies were similar in their antimicrobial susceptibility patterns. Nineteen strains were isolated from urine, four from faeces, two from wounds, nine from other human sources and two from animals.

Plant growth promotion and fungicidal activity in a siderophore-producing strain of Proteus sp.

A proteus strain inhibited mycelial growth of Fusarium oxysporum in vitro. Seed bacterization showed significant plant growth promotion and Fusarium-wilt suppression activity of Phaseolus mungo in a gnotobiotic system. The culture filtrate of this strain exhibited three prominent bands in UV-VIS spectra between 300 and 400 nm. The growth promotion assay of the extracted compound against different indicator organisms indicated the production of a compound related to a 2-oxoacid-type siderophore. The HPLC of the purified ethyl acetate extract of the strains and standard 4-methyl-2-oxopentanoate (2-oxoisocaproate) revealed a single peak, similarly as the coinjection of the extract and the standard. The production of siderophore, probably 2-oxoisocaproate, was demonstrated.

Thinking about bacterial populations as multicellular organisms.

It has been a decade since multicellularity was proposed as a general bacterial trait. Intercellular communication and multicellular coordination are now known to be widespread among prokaryotes and to affect multiple phenotypes. Many different classes of signaling molecules have been identified in both Gram-positive and Gram-negative species. Bacteria have sophisticated signal transduction networks for integrating intercellular signals with other information to make decisions about gene expression and cellular differentiation. Coordinated multicellular behavior can be observed in a variety of situations, including development of E. coli and B. subtilis colonies, swarming by Proteus and Serratia, and spatially organized interspecific metabolic cooperation in anaerobic bioreactor granules. Bacteria benefit from multicellular cooperation by using cellular division of labor, accessing resources that cannot effectively be utilized by single cells, collectively defending against antagonists, and optimizing population survival by differentiating into distinct cell types.

Potential virulence factors of Proteus bacilli.

The object of this review is the genus Proteus, which contains bacteria considered now to belong to the opportunistic pathogens. Widely distributed in nature (in soil, water, and sewage), Proteus species play a significant ecological role. When present in the niches of higher macroorganisms, these species are able to evoke pathological events in different regions of the human body. The invaders (Proteus mirabilis, P. vulgaris, and P. penneri) have numerous factors including fimbriae, flagella, outer membrane proteins, lipopolysaccharide, capsule antigen, urease, immunoglobulin A proteases, hemolysins, amino acid deaminases, and, finally, the most characteristic attribute of Proteus, swarming growth, enabling them to colonize and survive in higher organisms. All these features and factors are described and commented on in detail. The questions important for future investigation of these facultatively pathogenic microorganisms are also discussed.

Different biological conditions influencing bacterial adherence assay.

Adherence of bacteria to animal cells is considered the first step in the pathogenesis of many infectious diseases. The most suitable techniques developed in vitro to check the capacity of bacteria to adhere to different tissues use monolayers of established cell lines. We studied the influence of incubation time (1, 2, 3 hours), cell substrates (Hep-2, H-407) and the number of bacteria per cell (1, 10, 100, 1000) on the adherence index (number of adherent bacteria per cell determined by microscopic count) of the fimbriated Escherichia coli 454 strain, Proteus rettgeri 25 and Enterobacter cloacae 10. The data were analyzed with different statistical methods and the results evidenced that all the conditions considered affect either the end-point of the test or the adherence index. Our observations indicate that the different methods used make it impracticable to compare many data from the literature and suggest the need to search for more homogeneity in this type of assay.