Molecular Characterization of Escherichia coli O157:H7 and non-O157 Shiga Toxin Producing E. coli from Retail Meat and Humans

A total of 550 meat samples (300 minced beef and 250 chicken meat) marketed in Zagazig City, Sharkia Governorate, Egypt, as well as 150 human stool samples were examined for Shiga toxin producing E. coli. Results revealed that the isolation rates of E. coli O157:H7 versus non- O157 were 1.7% versus 2.3% in minced beef, 0.8% versus 2% in chicken meat and 0.7% versus 2.7% in human stools. Other identified serotypes were including O111:H8 (25%), O26:H11 (20.8%), O55:H7 (16.7%) and O113:H21 (4.2%). Virulence associated genes were identified in E. coli serotypes, stx1 and stx2 were characterized in 16.7% and 62.5% of the isolates, while, eaeA and hlyA genes were identified in 50% and 70.8% of the examined serotypes, respectively. Genotyping of E. coli O157:H7 serotype from different sources using Enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) fingerprinting revealed heterogenicity of the isolates, however, human and minced beef isolates were grouped in the same cluster indicating potential transmission of infection from contaminated beef to human consumers. In conclusion, ERIC-PCR is a highly discriminatory, reliable and cost-effective tool for tracing sources of infection with bacteria. Public health education and application of strict hygienic measures during slaughtering, transportation and preparation of meat are essential to minimize the risk of contamination and transmission of infection to consumer

Role of chitosan on controlling the characteristics and antifungal activity of bioadhesive fluconazole vaginal tablets

Vaginal fluconazole (FLZ) prolonged release tablets containing chitosan in physical blends with other bioadhesive polymers were designed. Chitosan was mixed with hydroxypropyl methylcellulose (HPMC), guar gum or sodium carboxymethyl cellulose (NaCMC) at different ratios and directly compressed into tablets. In-vitro release profiles of FLZ were monitored at pH 4.8. Compressing chitosan with HPMC at different ratios slowed FLZ release, however, time for 80% drug release (T80) did not exceed 4.3 h for the slowest formulation (F11). Adding of chitosan to guar gum at 1:2 ratio (F3) showed delayed release with T80 17.4 h while, in presence of PVP at 1:2:1 ratio (F5), T80 was 8.8 h. A blend of chitosan and NaCMC at 1:2 ratio (F15) showed prolonged drug release with T80 11.16 h. Formulations F5 and F15 showed fair physical characteristics for the powder and tablets and were subjected to further studies. Fast swelling was observed for F15 that reached 1160.53 ± 13.02% in 4 h with 2 h bioadhesion time to mouse peritoneum membrane compared with 458.83 ± 7.09% swelling with bioadhesion time exceeding 24 h for F5. Extensive swelling of F15 could indicate possible dehydration effect on vaginal mucosa. Meanwhile, antifungal activity against C. albicans was significantly high for F5

Design of taste masked enteric orodispersible tablets of diclofenac sodium by applying fluid bed coating technology

Diclofenac sodium (DS) a non-steroidal anti-inflammatory drug has a bitter taste and is a local stomach irritant. The aim of this study was to formulate taste masked DS orally dispersible tablets (ODTs) with targeted drug release in the intestine. Pellets of DS were designed using sugar sphere cores layered with DS followed by an enteric coat of Eudragit L100 and a second coat of Eudragit E100 for taste masking. The produced pellets had a high loading efficiency of 99.52% with diameters ranging from 493.7 to 638.9 µm. The prepared pellets were spherical with smooth surfaces on scanning electron microscopy examination. Pellets with the 12% enteric coat Eudragit L100 followed by 5% Eudragit E 100 resulted in 1.4 ± 0.5% DS release in simulated gastric fluid (SGF) and complete dissolution in simulated intestinal fluid (SIF). The pellets were then used to formulate ODTs. In vitro disintegration time of ODTs ranged from 20 ± 0.26 to 46 ± 0.27 s in simulated saliva fluid (SSF). Dissolution was less than 10% in SGF while complete drug release occurred in SIF. The release rate was higher for the optimized formulation (F12) in SIF than for the marketed product Voltaren® 25 mg tablets. The optimized ODTs formulation had a palatable highly acceptable taste. Keywords: Diclofenac sodium, Fluid bed coating, Enteric coated pellets, Enteric coated pellets, Taste masking, Orodispersible tablet

Optimization of 5-fluorouracil solid-lipid nanoparticles: a preliminary study to treat colon cancer

<p>Solid lipid nanoparticle (SLNs) formulae were utilized for the release of 5-fluorouracil (5-FU) inside the colonic medium for local treatment of colon cancer. SLNs were prepared by double emulsion-solvent evaporation technique (w/o/w) using triglyceride esters, Dynasan&#8482; 114 or Dynasan&#8482; 118 along with soyalecithin as the lipid parts. Different formulation parameters; including type of Dynasan, soyalicithin:Dynasan ratio, drug:total lipid ratio, and polyvinyl alcohol (PVA) concentration were studied with respect to particle size and drug entrapment efficiency. Results showed that formula 8 (F8) with composition of 20% 5-FU, 27% Dynasan&#8482; 114, and 53% soyalithicin and F14 (20% 5-FU, 27% Dynasan&#8482; 118, and 53% soyalithicin), which were stabilized by 0.5% PVA, as well as F10 with similar composition as F8 but stabilized by 2% PVA were considered the optimum formulae as they combined small particle sizes and relatively high encapsulation efficiencies. F8 had a particle size of 402.5 nm &#177; 34.5 with a polydispersity value of 0.005 and an encapsulation efficiency of 51%, F10 had a 617.3 nm &#177; 54.3 particle size with 0.005 polydispersity value and 49.1% encapsulation efficiency, whereas formula F14 showed a particle size of 343 nm &#177; 29 with 0.005 polydispersity, and an encapsulation efficiency of 59.09%. DSC and FTIR results suggested the existence of the lipids in the solid crystalline state. Incomplete biphasic prolonged release profile of the drug from The three formulae was observed in phosphate buffer pH 6.8 as well as simulated colonic medium containing rat caecal contents. A burst release with magnitudes of 26%, 32% and 28.8% cumulative drug released were noticed in the first hour samples incubated in phosphate buffer pH 6.8 for both F8, F10 and F14, respectively, followed by a slow release profile reaching 50%, 46.3% and 52% after 48 hours.</p

Development and Optimization of Ciprofloxacin HCl-Loaded Chitosan Nanoparticles Using Box&ndash;Behnken Experimental Design

Various chitosan (CS)-based nanoparticles (CS-NPs) of ciprofloxacin hydrochloride (CHCl) have been investigated for therapeutic delivery and to enhance antimicrobial efficacy. However, the Box&ndash;Behnken design (BBD)-supported statistical optimization of NPs of CHCl has not been performed in the literature. As a result, the goal of this study was to look into the key interactions and quadratic impacts of formulation variables on the performance of CHCl-CS-NPs in a systematic way. To optimize CHCl-loaded CS-NPs generated by the ionic gelation process, the response surface methodology (RSM) was used. The BBD was used with three factors on three levels and three replicas at the central point. Tripolyphosphate, CS concentrations, and ultrasonication energy were chosen as independent variables after preliminary screening. Particle size (PS), polydispersity index (PDI), zeta potential (ZP), encapsulation efficiency (EE), and in vitro release were the dependent factors (responses). Prepared NPs were found in the PS range of 198&ndash;304 nm with a ZP of 27&ndash;42 mV. EE and drug release were in the range of 23&ndash;45% and 36&ndash;61%, respectively. All of the responses were optimized at the same time using a desirability function based on Design Expert&reg; modeling and a desirability factor of 95%. The minimum inhibitory concentration (MIC) of the improved formula against two bacterial strains, Pseudomonas aeruginosa and Staphylococcus aureus, was determined. The MIC of the optimized NPs was found to be decreased 4-fold compared with pure CHCl. The predicted and observed values for the optimized formulation were nearly identical. The BBD aided in a better understanding of the intrinsic relationship between formulation variables and responses, as well as the optimization of CHCl-loaded CS-NPs in a time- and labor-efficient manner

Genetic Relatedness, Antibiotic Resistance, and Effect of Silver Nanoparticle on Biofilm Formation by Clostridium perfringens Isolated from Chickens, Pigeons, Camels, and Human Consumers

In this study, we determined the prevalence and toxin types of antibiotic-resistant Clostridium perfringens in chicken, pigeons, camels, and humans. We investigated the inhibitory effects of AgNPs on biofilm formation ability of the isolates and the genetic relatedness of the isolates from various sources determined using RAPD-PCR. Fifty isolates were identified using PCR, and all the isolates were of type A. The cpe and cpb2 genes were detected in 12% and 56% of the isolates, respectively. The effect of AgNPs on biofilm production of six representative isolates indicated that at the highest concentration of AgNPs (100 &micro;g/mL), the inhibition percentages were 80.8&ndash;82.8%. The RAPD-PCR patterns of the 50 C. perfringens isolates from various sources revealed 33 profiles and four clusters, and the discriminatory power of RAPD-PCR was high. Multidrug-resistant C. perfringens isolates are predominant in the study area. The inhibition of biofilm formation by C. perfringens isolates was dose-dependent, and RAPD-PCR is a promising method for studying the genetic relatedness between the isolates from various sources. This is the first report of AgNPs&rsquo; anti-biofilm activity against C. perfringens from chickens, pigeons, camels, and humans, to the best of our knowledge