Two-hundred-and-thirty-six isolates were collected from fresh flowers, bees and bee-hives. Of these, 20 isolates preferred D-fructose as carbon source, produced lactic acid and acetic acid but trace amounts of ethanol and were classified as fructophilic. Poor growth was recorded when strains were incubated anaerobically in the presence of D-glucose as sole carbon source. Good growth was, however, recorded when D-glucose was metabolized in the presence of external electron acceptors such as fructose, pyruvate and oxygen. Nineteen of the strains were classified as Lactobacillus kunkeei and one as Lactobacillus brevis based on phenotypic characteristics, 16S rRNA sequences, recA sequences and DNA homology. This is the first description of a fructophilic strain of L. brevis.
Copper particles were incorporated into nanofibers during the electrospinning of poly-D,L-lactide (PDLLA) and poly(ethylene oxide) (PEO). The ability of the nanofibers to prevent Pseudomonas aeruginosa PA01 and Staphylococcus aureus (strain Xen 30) to form biofilms was tested. Nanofibers containing copper particles (Cu-F) were thinner (326 ± 149 nm in diameter), compared to nanofibers without copper (CF; 445 ± 93 nm in diameter). The crystalline structure of the copper particles in Cu-F was confirmed by X-ray diffraction (XRD). Copper crystals were encapsulated, but also attached to the surface of Cu-F, as shown scanning transmission electron microscopy (STEM) and transmission electron microscopy (TEM), respectively. The copper particles had no effect on the thermal degradation and thermal behaviour of Cu-F, as shown by thermogravimetric analysis (TGA) and differential scanning calorimeter (DSC). After 48 h in the presence of Cu-F, biofilm formation by P. aeruginosa PA01 and S. aureus Xen 30 was reduced by 41% and 50%, respectively. Reduction in biofilm formation was ascribed to copper released from the nanofibers. Copper-containing nanofibers may be incorporated into wound dressings.
Silver nanoparticles (AgNPs), synthesized using N,N-dimethylformamide (DMF), were electrospun with nisin in a 50:50 blend of 24 % (w/v) poly(D,L-lactide) (PDLLA) and poly(ethylene oxide) (PEO). Addition of AgNPs decreased the average diameter of the nanofibers [silver nanofibers (SF)] from 588 ± 191 to 281 ± 64 nm, or to 288 ± 63 nm when nisin was co-spun with AgNPs. Nanofibers containing AgNO3 (SF) had a beads-on-string structure, whereas nanofibers with AgNPs and nisin [silver plus nisin nanofibers (SNF)], nanofibers with only nisin [nisin nanofibers (NF)], and nanofibers without AgNPs and nisin [control nanofibers] had a uniform structure. The irregular topography was confirmed by atomic force microscopy. No interactions occurred between silver, nisin, PDLLA, and PEO, as confirmed with Fourier transform infrared spectroscopy. Most of the AgNPs (18 ± 2.8 ppm) and nisin (78.1 ± 1.2 µg/ml) were released within the first 2 h. SF and SNF inhibited the growth of gram-positive and gram-negative bacteria, whereas NF failed to inhibit gram-negative bacteria. A wound dressing with broad-spectrum antimicrobial activity may be developed by the incorporation of nanofibers containing a combination of AgNPs and nisin.
Pseudomonas aeruginosa and Staphylococcus aureus are commonly associated with hospital-acquired infections and are known to form biofilms. Ciprofloxacin (CIP), which is normally used to treat these infections, is seldom effective in killing cells in a biofilm. This is mostly due to slow or weak penetration of CIP to the core of biofilms. The problem is accentuated by the release of CIP below MIC (minimal inhibitory concentration) levels following a rapid (burst) release. The aim of this study was to develop a drug carrier that would keep CIP above MIC levels for an extended period. Ciprofloxacin was suspended into poly(D,L-lactide) (PDLLA) and poly(ethylene oxide) (PEO), and electrospun into nanofibers (CIP-F). All of the CIP was released from the nanofibers within 2 h, which is typical of a burst release. However, 99% of P. aeruginosa PA01 cells and 91% of S. aureus Xen 30 cells (a methicillin-resistant strain) in biofilms were killed when exposed to CIP-F. CIP levels remained above MIC for 5 days, as shown by growth inhibition of the cells in vitro. The nanofibers were smooth in texture with no bead formation, as revealed by scanning electron and atomic force microscopy. A single vibration peak at 1632 cm-1, recorded with Fourier transform infrared spectroscopy, indicated that CIP remained in crystal form when incorporated into PDLLA: PEO. No abnormalities in the histology of MCF-12A breast epithelial cells were observed when exposed to CIP-F. This is the first report of the inhibition of biofilm formation by CIP released from PDLLA: PEO nanofibers.
Antibiotics in feed select for resistant strains and is thus a threat to human health. In this study, the effect of a multi-strain probiotic and antibiotics on the growth and health of broilers was studied. Equal numbers of broilers received on a daily basis either a multi-strain probiotic or a combination of sulphadiazine, colistin and trimethoprim, whereas the control group received standard feed. The villi of immature broilers (19 days old) administered antibiotics had a larger surface area and their lymphocyte and basophil counts were higher compared to broilers from the probiotic and control groups. The cecal microbiomes of mature broilers (29 days old) that received probiotics had higher levels of Enterobacteriaceae, but lower numbers of Clostridiales, Brucellaceae, Synergistaceae, Erysipelotrichaceae and Coriobacteriaceae compared to the antibiotic-treated group. A decline in the bioluminescence of Listeria monocytogenes observed for broilers on probiotics suggested that the probiotic may be used to control bacterial infections. No significant differences in total red blood cell, haemoglobin and haematocrit content, and mean values for corpuscular volume, corpuscular haemoglobin and corpuscular haemoglobin numbers were recorded amongst broilers from the different treatment groups. This study provides valuable information on the health and performance of broilers when administered probiotics and antibiotics as additives.
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