Isolation And Identification Of Rhodotorula Mucilaginosa TZR₂₀₁₄ And Its Application In Weaned Piglets

The purpose of this experiment mainly isolates Rhodotorula mucilaginosa from the different fruit soil in Chongqing Province.Then using Rhodotorula mucilaginosa to identification of safety tests.It was studied by feeding the weaned piglets experiment.The effectiveness of probiotics on piglets production performance,serum biochemical parameters,pH value of the gastrointestinal tract,organ index,digestive enzymes activities,intestinal mucosal morphology and intestinal flora were used as evaluated index.Trial 1 was isolated Rhodotorula mucilaginosa from the different fruit soil in Chongqing Province.The isolate was identified based on the characteritics of cology morphology.Then identified it through physio logical biochemical method and 26 S rDNA D1/D2 of sequence analysis.The result showed the yeast is Rhodotorula mucilaginosa.Trial 2 conducted the safety evaluation for Rhodotorula mucilaginosa in the experimental model,which include acute toxic ity text,ectopic textsperm.20 rats were randomly divided into 2 groups(10 rats per group): CON group orally administrated 0.1 mL saline;Rhodotorula mucilaginosa group orally administrated 0.1 mL1.0×1010 Rhodotorula mucilaginosa fermentation broth,continued for 3 days.Acute toxicological tests showed that Rhodotorula mucilaginosa will not cause adverse reactions in the signs of mice,behavior,activities,hair color,diet and excrement.There was no significant difference in body weight campare with the control grop(P>0.05).Pathological changes were not found of major organs such as the liver,spleen and kidneys.40 rats were randomly divided into 4 groups(10 rats per group): CON group orally administrated 0.1 mL saline;group 2 orally administrated 0.1 mL 1.0×107;group 2 orally administrated 0.1 mL 1.0×108;group 2 orally administrated 0.1 mL 1.0×109,continued for 30 days.During the experiment observed in the signs of mice,behavior,activities,hair color,diet and excrement.The end of the experiment,tomeasure the weight of body,liver,spleen and conduct ectopic trals.The result showed anomalies in the major oagan of mice,included the liver and spleen.The organs of the mice culture on PDA medium does not appear coloines of yeast limplied that there was no yeast ectopic phenomenon.So we can preliminarily determine that the use of Rhodotorula mucilaginosa.4 groups mice will not cause adverse reactions in the signs of mice,behavior,activities,hair color,diet and excrement.Group 4 the weight of body,liver and spleen compare with other grpups was significant(P<0.05),group 2 and group 3 the weighe of body,liver and spleen campare with group 1 was no significant(P>0.05).The organ of the mice culture on PDA medium does not appear coloines of Rhodotorula mucilaginosa limplied that there was no yast ectopic phenomenon.So we can peliminarily determine that the use of Rhodotorula mucilaginosa.Rhodotorula mucilaginosa acid,bile salt,high temperature drug sensitive resistance results showed that Rhodotorula mucilaginosa can acid,bile salt resistance.The survival rates drop dramatically after 65 ?.It is not sensitive to penicillin and sensitive to chloramphenicol,Achromycin,kanamycin and Gentamicin.Trial 3 was conducted to investigate the effect of dietar Rhodotorula mucilaginosa on growth performance,organ index,plasma parameters,intestinal mucosa morphology,digestive enzymes activities and microflora of colon in weaned piglets.30 weaned piglets were randomly divided into three groups(10 piglets per group): control proup(CON),R.mucilaginosa group,C.utilis group.The experiment lasted 35 d including 7 d adaptation.The final weight,average daily and dry matter intake of weaned piglets in R.mucilaginosa group(P<0.05).Compare with other groups,feed conversion and rate of diarrhea of piglets in R.mucilaginosa group was decreased(P<0.05).The heart index,liver index,thymus index,kidney index and pancreas index of piglets in CON group,R.mucilaginosa group and C.utilis group was no significant(P>0.05).The spleen index and lymph node index were higher than in CON group and C.utilis group.The plasma concentration of alkaline phosphatase and nitric oxide synthase of piglets was no significant.The plasma concentration of total antioxidant capacity and lysozyme of piglets in R.mucilaginosa group were higer than CON group(P<0.05).The plasma concentration of total superoxide sismutase and glutathione of piglets in R.mucilaginosa group were higher than that in CON and C.utilis groups.The plasma concentration of malonic dialdehyde of piglets in R.mucilaginosa group was lower than in CON and C.utilis groups(P<0.05).The jejunum and ileun lenhth index of piglets in three groups was no significant(P<0.05).The weight of jejunum and ileun in R.mucilaginosa group were higher than that in CON and C.utilis groups(P<0.05).The pepsin activity in jejunum of R.mucilaginosa group was improved compare with CON group(P<0.05).There was no significant difference amone each treatment groups of amylase activity(P>0.05).The R.mucilaginosa group increase the activity of lipase in jejunum than that in CON and C.utilis groups(P<0.05).There was no significant difference amone each treatment groups of stomach pH-value(P>0.05).The jejunum pH-value of R.mucilaginosa group was lower compare with other groups(P<0.05).The ileum p H-value,colon pH-value,cecum pH-value were significant difference in R.mucilaginosa group campare with CON group(P<0.05).The number of proteobacteria,actinobacteria in R.mucilaginosa group was significant higher than it in CON and C.utilis groups(P<0.05).The number of Bacteroiddetes,Proteobacteria,Actinobacteria in R.mucilaginosa group was lower than it in CON group(P<0.05).The number of Firmicutes,Verrucomicrobia and Spirochaetae was significant difference in R.mucilaginosa group.There were no significant difference of abundance of the colony and diversity of species amone each treatment group.General conclusions can be drawn based on the above results: R.mucilaginosa can improve the gowth performance,enhance the vitality of digestive enzymes,maintain the intestinal microflora balance.Thereore,as a natural additives,R.mucilaginosa can be applicable to feed production of animal.