Effect of some antibiotics and alcoholic extracts of Loranthus europaeus on growth of Helicobacter pylori by using nanochitosan

 This study was aimed to an attempt for early detection of Helicobacter pylori bacterium by relying the relationship of ammonia production with changing the color of the media, pH, bacterial growth density and the ulceration type (acute or hemorrhage). The study also aimed to use biodegradable synthetic nanomaterials as a mean of   delivery for therapy as well as knowledge of their efficiency as a therapeutic agent. H. pylori considered as the most causative agent of stomach and duodenum ulcer which may develop into stomach or duodenum cancer.  New broth medium (ZAN-1) and modified media (Brain Heart Infusion BHI, and Tryptic Soybean Broth, TSB) were used to stimulate and support bacterial growth since it is difficult to be grown and isolated. Results of selection acute and hemorrhage isolates were based on ammonia production within 2 hours and changing the color of fast urea solution in addition to the histological tests, immunological tests, confirmation tests and genomic profile. A nano carrier from chitin were developed to produce chitosan by ionic gelation method and used it as carrier and therapeutic agent. Results  obtained could be summarized as follows:
 A total of 115 biopsies specimens were collected  from gastroscopy department in Alkadumia hospital during June to September,2014 from adult aged from 40 to 60 years. Eighty of these samples for males and 35 for females, Ten isolates were selected for each acute and hemorrhage infection depending on the direct tests in hospital; and based on the laboratory and hospital tests , one acute isolate (A-1) and one hemorrhage isolate (H-1) were selected.Genomic profile study revealed that acute and hemorrhage isolates (A-1, H-1) of H.pylori have only genomic DNA and absence of plasmids.Susceptibility test showed that A-1 was less response to antibiotics than H-1. Phenate method was selected to be the best method to estimate the concentrations of ammonia in culture media of this bacterium, results indicated that acute isolate is more active in ammonia production (0.41µg/ml) and changing the color of media comparing with hemorrhage isolate which was (0.38 µg/ml) ammonia. Prepare nanochitosan and test its shape using SEM.Prepare of Loranthus europaeus extract. Preparation of nanochitosan loaded with antibiotic and L. europaeus extract and prepare lyophilized mixtures. Semi- quantitative applications and results showed clear superiority thatA-1 isolate was more sensitive to the nanachitosan  loaded with the antibiotic (ciprofloxacin) and give inhibition zone 48 mm after 48 hr incubation, while when used ciprofloxacin alone it give15 mm inhibition after 48 hr of incubation. Quantitative applications illustrated that the higher  concentration of ammonia production could be detected by spectrophotometer  was 5.2 µg/ml for A-1 isolate using   inoculm  3×10 5 cell/ml  for nanochitosan loaded with ciprofloxacin. Invitro application showed that the nanochitosan with ciprofloxacin have more inhibitory action on the cells. (Ammonia production was 4.5µg /ml compared with the ciprofloxacin alone 41µg/ml).