Studies On Antibacterial Components From Maggot Homogenate And Its Enzymatic Hydrolysate

This study determined the effect of maggot homogenate on the growth of E.coli,Samonella,Staphylococcus aureus and Bacillus subtilis, elucidated anti-infection effect oftissue homogenate on E.coli and Samonella in mice; hydrolyzed the tissue homogenate byusing trypsin and pepsin, determined the effect of enzymatic hydrolasate on the growth offour strains, elucidated anti-infection effect of tissue homogenate on E.coli and Samonellain mice;and then separated the antibacterial components from tryptic hydrolysate by usingSephadexG-15 and RP-HPLC.1. The effect of maggot homogenate on the growth of strains andanti-infection effect on Escherichia.coli and Samonella in miceA sort of actively antibacterial components was obtained from the homogenate ofmaggot treated with phosphate buffer solution. The results indicated that the homogenate ofmaggot exhibits significantly antibacterial activity to Escherichi coli and Samonella, andaccelerates the growth of Staphylococcus aureus and Bacillus subtilis,but the antibacterialactivity disappeared when heated at 100℃. The second part of this study was to elucidateanti-infection effect of maggot homogenate on E.coli and Sarnonella in mice. According totoxicity test, determinate the dosage of E.coli and Samonella in mice. Two batches of 60Male KM mice were randomly assigned to one of three treatments(n=20): normal group(feeding-E.coli control group):Basal diet + physiological saline, Treatment control group:Basal diet + maggot homogenate that heated at 100℃, Treatment group: Basal diet+maggot homogenate.Feeding level of every mouse was 0.3mL with equal amount ofvolume. After fed 7 days, every mouse from Treatment control group to Treatment groupwas fed E.coli or Samonella in the 8th day midday. The feeding level of every mouse is0.3 mL with E.coli 1.8×10¹¹ CFU/L and Samonella 2.7×10¹¹ CFU/L respectively. Observethe healthy of mice in each group until feeding E.coli or Samonella after 56 h, kill the mice,collect chyme from cecum, and using Colony-forming unit (CFU) assay to determine theamount of special strain. The results illustrated that KM mice reacted differently when infected E.coli or Samonella after orally administrated with the maggot homogenate. Themaggot homogenate could significantly increase the level of health (E.coli Treatmentgroup increased 83.25% vs Treatment control group; Samonella Treatment groupincreased 42.85% vs Treatment control group), decrease the amount of special strain inchyme from cecum, (Treatment group vs Treatment control group, p＜0.05). The resultsdemonstrated that maggot homogenate could exhibits significantly antibacterial activity toEscherichicoli and Samonella and increase the immune function in mice, defend theinvasion of E.coli and Samonella, and keep the gut health.2. The effect of enzymatic hydrolysate on the growth of strains andanti-infection effect on Escherichia.coli and Samonella in miceAfter hydrolyzed by trypsin and pepsin, and then effect the growth of strains, Theresults indicated that the tryptic hydrolysate exhibits significantly antibacterial activity toEscherichicoli and Samonella, and accelerates the growth of Staphylococcus aureus andBacillus subtilis, without obvious chang in antibacterial activity after treated at 100℃;peptic hydrolysate accelerates the growth of all four strains. The second part of this studywas to elucidate anti-infection effect of maggot homogenate on E.coli and Samonella inmice. According to toxicity test, determination the dosage of E.coli and SamonellaI in mice.Two batches of 50 Male KM mice were randomly assigned to one of threetreatments(normal group:10,the other two treatments: 20): normal group:Basal diet +physiological saline, Treatment control group: Basal diet + physiological saline, Treatmentgroup: Basal diet + enzymatic hydrolysate. Feeding level of every mouse was 0.3mL withequal amount of volume. After fed 7 days, every mouse from Treatment control group toTreatment group was fed E.coli or Samonella in the 8th day. The feeding level of everymouse is 0.3mL with E.coli 3.38×10¹¹ CFU/L and Samonella 2.87×10¹¹ CFU/L respectively.Observe the healthy of mice in each group until feeding E.coli or Samonella after 56h, killthe mice, collect chyme from cecum, and using Colony-forming unit (CFU) assay todetermine the amount of special strain. The results illustrated that KM mice reacteddifferently when infected E. coli or Samonella after orally administrated with the enzymatichydrolysate. The tryptic hydraolydate could significantly increase the level of health (E.coliTreatment group increased 25.02% vs Treatment control group; Samonella Treatmentgroup increased 14.27% vs Treatment control group), decrease the amount of special strainin chyme from cecum, (Treatment group vs Treatment control group, p＜0.05). The resultsdemonstrated that tryptic hydrolysate could exhibits significantly antibacterial activity to Escherichicoli and Samonella and increase the immune function in mice, defend theinvasion of E.coli and Samonella, and keep the gut health.3. The separation of antibacterial components from tryptichydrolysisWhen isolated with Sephadex G-15 after tryptic hydrolysate, we obtained threeabsorption peaks. The results indicated that components from the third absorption peakexhibit significantly antibacterial activety to Escherichia coli and Samonella butaccelerate the growth of Staphylococcus aureus and Bacillus subtilis, and thecomponents from the other two absorption peaks accelerate the growth of all four strains.And by usingRP-HPLC method, four main peaks were separated from the third absorptionpeak. The experimental proof could be deduced that antibacterial substances existed inthe tryptic hydrolysate.