Effects of Chloride Concentration on the Water Disinfection Performance of Silver Containing Nanocellulose-based Composites.

The availability of microbially-safe drinking water is a challenge in many developing regions. Due to the well-known antibacterial effect of silver ions, materials used for their controlled release have been widely studied for point-of-use water disinfection. However, even if it is in principle known that chloride anions can suppress the antibacterial efficiency of silver, the majority of previous studies, surprisingly, have not focused on chloride concentrations relevant for freshwaters and thus for practical applications. Here, we prepared low-cost nanocellulose-aluminium oxyhydroxide nanocomposites functionalized with silver nanoparticles. Field samples obtained from Chennai, India were used as a guideline for choosing relevant chloride concentrations for the antibacterial studies, i.e., 10, 90, and 290 ppm. The antibacterial performance of the material against Escherichia coli and Bacillus subtilis was demonstrated and the influence of chloride concentration on the antibacterial effect was studied with E. coli. A 1 h contact time led to bacterial reductions of 5.6 log10, 2.9 log10, and 2.2 log10, respectively. This indicates that an increase of chloride concentration leads to a substantial reduction of antibacterial efficiency, even within chloride concentrations found in freshwaters. This work enables further insights for designing freshwater purification systems that utilize silver-releasing materials.

Impact of diets with a high content of greaves-meal protein or carbohydrates on faecal characteristics, volatile fatty acids and faecal calprotectin concentrations in healthy dogs.

BACKGROUND: Research suggests that dietary composition influences gastrointestinal function and bacteria-derived metabolic products in the dog colon. We previously reported that dietary composition impacts upon the faecal microbiota of healthy dogs. This study aims at evaluating the dietary influences on bacteria-derived metabolic products associated with the changes in faecal microbiota that we had previously reported. We fed high-carbohydrate starch based (HCS), [crude protein: 194 g/kg, starch: 438 g/kg], high-protein greaves-meal (HPGM), [crude protein: 609 g/kg, starch: 54 g/kg] and dry commercial (DC), [crude protein: 264 g/kg, starch: 277 g/kg] diets, and studied their effects on the metabolism of the colonic microbiota and faecal calprotectin concentrations in five Beagle dogs, allocated according to the Graeco-Latin square design. Each dietary period lasted for three weeks and was crossed-over with washout periods. Food intake, body weight, and faecal consistency scores, dry matter, pH, ammonia, volatile fatty acids (VFAs), and faecal canine calprotectin concentrations were determined. RESULTS: Faecal ammonia concentrations decreased with the HCS diet. All dogs fed the HPGM diet developed diarrhoea, which led to differences in faecal consistency scores between the diets. Faecal pH was higher with the HPGM diet. Moreover, decreases in propionic and acetic acids coupled with increases in branched-chain fatty acids and valeric acid caused changes in faecal total VFAs in dogs on the HPGM diet. Faecal canine calprotectin concentration was higher with the HPGM diet and correlated positively with valeric acid concentration. CONCLUSIONS: The HPGM diet led to diarrhoea in all dogs, and there were differences in faecal VFA profiles and faecal canine calprotectin concentrations.

Effect of high contents of dietary animal-derived protein or carbohydrates on canine faecal microbiota.

BACKGROUND: Considerable evidence suggests that food impacts both the gastro-intestinal (GI) function and the microbial ecology of the canine GI tract. The aim of this study was to evaluate the influence of high-carbohydrate (HC), high-protein (HP) and dry commercial (DC) diets on the canine colonic microbiota in Beagle dogs. Diets were allocated according to the Graeco-Latin square design. For this purpose, microbial DNA was isolated from faecal samples and separated by density gradient centrifugation, resulting in specific profiling based on the guanine-cytosine content (%G+C). In addition, 16 S rRNA gene amplicons were obtained from the most abundant %G + C peaks and analysed by sequence analysis, producing a total of 720 non-redundant sequences (240 sequences per diet). RESULTS: The DC diet sample showed high abundance of representatives of the orders Clostridiales, Lactobacillales, Coriobacteriales and Bacteroidales. Sequence diversity was highest for DC diet samples and included representatives of the orders Lactobacillales and Bacteroidales, which were not detected in samples from the HP and HC diets. These latter two diets also had reduced levels of representatives of the family Lachnospiraceae, specifically Clostridial cluster XIVa. The HC diet favoured representatives of the order Erysipelotrichales, more specifically the Clostridial cluster XVIII, while the HP diet favoured representatives of the order Fusobacteriales. CONCLUSIONS: This study detected Coriobacteriales in dog faeces, possibly due to the non-selective nature of the %G + C profiling method used in combination with sequencing. Moreover, our work demonstrates that the effect of diet on faecal microbiota can be explained based on the metabolic properties of the detected microbial taxa.

Characterization of microbial contaminants in urine.

Modern, molecular microbiological methods were applied to urine samples from control subjects and athletes for characterization of the microbial community. High abundance of lactobacilli, enterococci, and enterobacteria was detected in urine samples, suggesting that gastrointestinal and urogenital tracts act as contamination sources. Athlete samples, but not control samples, showed an abundance of pseudomonads, a bacterial group reported to metabolize steroids. Overall, the bacteria detected are known to be capable of altering steroid profiles, emphasizing the importance of good hygiene at sampling in reliable doping control.

Combination of polydextrose and lactitol affects microbial ecosystem and immune responses in rat gastrointestinal tract.

The effects of various dietary fibres on gut health have been studied extensively but their combined effects are scarcely documented. In the present study the effects of 2 % (w/w) polydextrose (PDX), 2 % (w/w) disaccharide lactitol, or 2 % (w/w) PDX+2 % (w/w) lactitol on gut microflora, microbial metabolism and gut immune responses were investigated in rats. Both PDX and lactitol alone had an effect on many of the studied parameters, but their combination had stronger than additive effects in some parameters. The PDX+lactitol combination altered the microbial community structure as seen by a culture-independent method, percentage guanine+cytosine (%G+C) profiling, increasing the areas of %G+C 35-39 (P<0.0001) and %G+C 45-49 (P=0.0002), and decreasing %G+C 65-74 (P<0.0003). These changes were also reflected in the microbial metabolism so that the production of biogenic amines and branched volatile fatty acids was significantly reduced, by 12 (P=0.03) and 50 % (P=0.002), respectively, indicating a shift from putrefactive towards saccharolytic metabolism. PDX increased the secretion of IgA in the caecum (P=0.007). Secretion of IgA increased even more, almost ten-fold, with the combination of PDX+lactitol (P<0.0001) when compared with the control group. Lactitol increased the production of butyrate by caecal microbes by two- to three-fold when compared with the PDX or control group (P<0.0001). Butyrate is a preferred energy source for mucosal cells; thus a boost in the availability of energy for immune cells may have still added to the synergistic effects of PDX and lactitol on immune cells. It is noteworthy that improvement in the IgA secretion occurred without signs of mucosal inflammation.

GC fractionation enhances microbial community diversity assessment and detection of minority populations of bacteria by denaturing gradient gel electrophoresis.

Effectively and accurately assessing total microbial community diversity is one of the primary challenges in modern microbial ecology. This is particularly true with regard to the detection and characterization of unculturable populations and those present only in low abundance. We report a novel strategy, GC fractionation combined with denaturing gradient gel electrophoresis (GC-DGGE), which combines mechanistically different community analysis approaches to enhance assessment of microbial community diversity and detection of minority populations of microbes. This approach employs GC fractionation as an initial step to reduce the complexity of the community in each fraction. This reduced complexity facilitates subsequent detection of diversity in individual fractions. DGGE analysis of individual fractions revealed bands that were undetected or only poorly represented when total bacterial community DNA was analyzed. Also, directed cloning and sequencing of individual bands from DGGE lanes corresponding to individual G+C fractions allowed detection of numerous phylotypes that were not recovered using a traditional random cloning and sequencing approach.

Selective plating underestimates abundance and shows differential recovery of bifidobacterial species from human feces.

The aim of the present work was to compare the efficacies and levels of selectivity of different culture-dependent and -independent methods for analyzing bifidobacteria in human stool samples. The three different culture media used here significantly differed from each other, particularly with regard to the recovery of Bifidobacterium adolescentis. Bifidobacterium medium failed to recover B. adolescentis; Beerens medium recovered some B. adolescentis organisms (17% of total bifidobacteria), whereas tomato-Eugon medium recovered mainly B. adolescentis organisms (58% of total bifidobacteria). A culture-independent method that combines GC fractionation of bacterial community DNA and 16S rRNA sequencing indicated that B. adolescentis organisms accounted for 85% of all bifidobacteria. Methodological biases, such as those described in this paper, should be taken into account in interpreting earlier studies and designing future experiments.

Intestinal immune responses in wild-type and Apcmin/+ mouse, a model for colon cancer.

C57BL/6J Apc(Min/+) (Apc(Min/+)) mice spontaneously develop pretumoric adenomas into the intestinal mucosa. We studied the relationship between the intestinal immune responses and adenoma formation in Apc(Min/+) mice and compared Apc(Min/+) mice with their wild-type siblings. Three time points (5, 8, and 15 weeks of age) and three high-fat dietary treatments (a non-fiber control with beef or inulin amendment) were included. The numbers of CD8+ T lymphocytes and tissue macrophages (Mac-1+ cells) per villus in ileal mucosa were determined by immunohistology, and the concentrations of secretory IgA, residual prostaglandin E(2) (PGE(2)), tumor necrosis factor alpha, and interleukin (IL)-12 in ileal contents were analyzed by ELISA. The crypt-villus ratio of the ileal mucosa was determined histologically. An immunostimulation, characterized by an increase in several parameters (PGE(2), IgA, Mac-1, and CD8), was observed in both genotypes between weeks 5 and 8. Most of the adenomas in Apc(Min/+) mice also appeared during the same period of sexual maturation. Females had smaller adenomas than males, and the beef group had fewer and smaller adenomas than the control group at 15 weeks. Females had less IgA and fewer Mac-1+ and CD8+ cells in ileal tissue than males at 15 weeks and more luminal IL-12 than males at 8 weeks. Puberty may have affected both tumorigenesis and intestinal immune responses in the Apc(Min/+) mouse. The beef group showed less luminal IgA and tumor necrosis factor alpha but more IL-12 than the control group. The concentration of PGE(2) correlated positively with the number and size of adenomas and was higher in the Apc(Min/+) mice than in wild-type mice at 15 weeks. IgA and Mac-1 were positively correlated with the size of adenomas at 15 weeks. The positive correlations between tumor size and IgA and between tumor number and size and PGE(2) suggest that a balance toward the Th2 type immune response may affect the pace of tumorigenesis in this model. The general similarity of the intestinal immune responses in both genotypes and the lack of intestinal inflammation in the Apc(Min/+) mice suggest that the mutation in the adenomatous polyposis coli gene does not lead to major alterations in intestinal immune function and that the intestinal immunology does not explain tumorigenesis in the Apc(Min/+) mouse model.

Characterization of poultry egg-white avidins and their potential as a tool in pretargeting cancer treatment.

Chicken avidin and bacterial streptavidin are proteins used in a wide variety of applications in the life sciences due to their strong affinity for biotin. A new and promising use for them is in medical pretargeting cancer treatments. However, their pharmacokinetics and immunological properties are not always optimal, thereby limiting their use in these applications. To search for potentially beneficial new candidates, we screened egg white from four different poultry species for avidin. Avidin proteins, isolated from the duck, goose, ostrich and turkey, showed a similar tetrameric structure, similar glycosylation and stability against both temperature and proteolytic activity of proteinase K as chicken avidin. Biotin-binding properties of these avidins, measured using IAsys optical biosensor, were similar to those found in avidin from the chicken. Three of these novel avidins, however, showed different immunological cross-reactivities when compared with chicken avidin. The patient sera responses to duck, goose and ostrich avidins were also lower than those observed for chicken and turkey avidins. Our findings suggest that the use of these proteins offers advantages over chicken avidin and bacterial streptavidin in pretargeting applications.

Culture-independent microbial community analysis reveals that inulin in the diet primarily affects previously unknown bacteria in the mouse cecum.

Inulin is a well-known fructose-based prebiotic which has been shown to stimulate the growth of bifidobacteria, a bacterial group generally considered beneficial for intestinal health. In the present study, we analyzed inulin-associated shifts in the total bacterial community of wild-type mice and mice carrying a genetically inactivated adenomatous polyposis coli tumor suppressor gene by using DNA-based approaches independent of bacterial culturability. Mice were fed a high-fat, nonfiber diet with or without inulin inclusion at a 10% (wt/wt) concentration. Cecal contents were analyzed after 0, 3, and 9 weeks on the experimental diets. Inulin inclusion significantly affected the total bacterial community structure of the cecum as determined by both a nonselective percent-guanine-plus-cytosine-based profiling analysis and a more specific 16S ribosomal DNA sequence analysis. The shifts included stimulation of bifidobacteria and suppression of clostridia, but sequence comparison revealed that the major shifts were within previously unknown bacterial taxa. Concomitantly, significantly higher bacterial densities, determined by flow cytometry, were observed with the inulin-amended diet, and the metabolism of the cecal bacterial community was altered, as indicated by higher levels of residual short-chain fatty acids, particularly lactic acid. With regard to all of the microbiological parameters measured, the wild-type mice and mice carrying a genetically inactivated adenomatous polyposis coli tumor suppressor gene were essentially identical. Studies of the implications of pre- and probiotics may need to be expanded to include careful analysis of their effects on the entire microbial community, rather than just a few well-known species. Further studies are needed to increase our understanding of the possible roles of currently unknown gastrointestinal bacteria in health and disease.