تاثير السكر المتعدد الخارجي والخلايا المقتولة حراريا لعزلات محلية من بكتريا حامض اللاكتيك في تثبيط نمو الخلايا السرطانية وكمضادات اكسده ومايكروبيه == Effect of the expolysacaccarid and heat killed cells for local isolates of Lactic acid bacteria in growth inhibition of cancer cells and as antioxidation and antimicrobial
Author name:
فاطمة حسن احمد اللعيبي
Supervisor name:
قيثار رشيد مجيد | ناهي يوسف ياسين
General topic:
Agricultural sciences
Specific topic:
Food
Degree:
Master
University:
University Of Basrah - Faculty Of Agriculture
Language:
Arabic
University location:
Basrah
First pages:
31T1843 - p.pdf
Abstract:
Lactobacillus bacteria was Isolated from different food samples included (bananas, tomatoes, apples, mandarin orange, cheese, buffalo milk, local yogurt and imported yogurt) and from infant feaces . 65 isolates were obtained out of 64 food samples, and 11 samples of infant feaces belong to Lactobacillus spp. and identified depending on selective medium MRS agar as well as phenotypic tests, microscopic tests and biochemical examinations including catalase test, production of Co2 from glucose, gelatin liquefaction, ammonia reduction from arginine, nitrate reduction to nitrite, growth examination in different temperatures, indole test, salinity tolerance test, motility test and carbohydrate fermentation test. As well as, confirmed test was done by using PCR technique. Purification and identification conducted and showed it contain (20) isolates of Lactobacillus acidophilus, (23) isolates of Lactobacillus plantarum, (6) isolate of Lactobacillus casei, (12) isolates of Lactobacillus bulgaricus, (3) isolates of bacteria Lactobacillus johnsonii and one isolates of Lactobacillus helveticus. The identified isolates of Lactic acid bacteria were tested on the production of polysaccharide and the optimal conditions for the production by using the modified medium by adding lactose to the medium and vaccine volume was 0.1% and the temperature of incubating was 37c° for (24 - 48) hrs and pH was (6.2 - 6.5) and centrifuged (11000) rpm. Detction of polysaccharide for all obtained bacteria was done . Results showed that the bast of amount production of polysaccharide to(Lactobacillus plantarum )which isolate from imported yoghurt (yR5) because it got the best amount of polysaccharide production which was (850.35) m / ml.SummaryThe presence of polysaccharide was detected by using thin - layer chromatography technique TLC to make sure from the presence of sugar molecules by using sulfuric acid and then the produced polysaccharide was detected by using High Performance Liquid Chromatography Technology HPLC with the existence of standard saccharides for the purpose of detection and comparison.Inhibition activity of both crude bacteria cells and produced polysaccharide were studied against gram positive and negative bacteria which included Staphylococcus aureus, Escherichia coli, Streptococcus spp., Pseudomonandas aeruginosa and E. coli O157 : H7 as well as the inhibitory activity against fungi was determined for each crude bacteria cells and produced polysaccharide especially against Penicillium spp. and Asprgills niger Result showed that in habithion effect was apparel towel all microorganism however no effect E. coli O157 : H7.Anti - oxidant activity of crude bacterial isolates were tested and the best anti - oxidant activity (Lactobacillus acidophilus (was the isolate which isolated from bananas source (B4) Mar 51.01% reached while less effective was the isolation (Lactobacillus bulgaricus) isolated from the local yogurt source (yL2) if it reaches 14.04% and the highest antioxidant activity of polysaccharide was from the isolate(Lactobacillus bulgaricus) which isolated from the local yogurt source (yL6) , amounting to 67.51% while it was less effective than isolation (Lactobacillus acidophilus ) isolated from the local chess source (Cha2) as it was 1.29% compared with the industrial anti - oxidation BHT and the natural anti - oxidation α - tocopherol in concentration (0.05 g / ml), results showed that the anti - oxidant activity by using the industrial anti - oxidant BHT reached to 82.70%, while the highest inhibitory activity of bacteriaSummaryanti - oxidant compounds 51.01% and for polysaccharide the highest percentage of inhibition was 67.51%, the bacteria anti - oxidant compounds and polysaccharide activity were close, while the activity of the natural anti - oxidants α - tocopherol was 73.96%.The results of sensitivity toward 10 antibiotics showed that all isolates of lactobacillus spp. Were sensitive to clindamycin. While only 16% of isolates were sensitive to nalidaxic acid.The cytotoxicity effect of the Heat killed (HK) cells and polysaccharide extract (EPS) of local isolates of Lactobacillus spp. on three of the cancer cells lines, human cervix uteri epitheloid carcinoma (Hela), muscular cancer cells line (RD) and rat embryogenic fibroblast (REF) as normal cell line , were studied . In this study, six concentrations of both polysaccharide extract and the (HK) cells which were (93.5, 187.5 , 375, 750,1500, 3000) mcg/ml. The results showed that all types of dead cells of lactic acid bacteria and polysaccharide product which clear effect on tumor cells differed depending on the degree of influence, polysaccharide extract of Lactobacillus johnsonii showed highest percentage of inhibition and the highest percentage of inhibition for (Hela) line was (93.64%) at exposure time of 72 hours. while the highest percentage of inhibition recorded for Lactobacillus acidophilus in (RD) line cells was (90.63%) after 72 hours. For the (HK) cells, the highest percentage of inhibition for Lactobacillus casei was 89.77 after 48 hours. While it was no effect of polysaccharide and HK on (Rat Embroy Fibroblast) ofter exposure time 72h.
