| Chinese herbal medicines are natural herbal medicine made of additives,so the production of animal products is natural without the addition of toxic and harmful substances.Mycoplasma Synoviae is an acute or chronic infectious disease characterized by exudative synovial bursitis and tenosynovitis in chickens,ducks,geese and wild birds such as cranes,quails and pigeons.The application of Chinese herbal medicine has provided more good quality livestock products for people.So,keeping in view the risk of MS and the importance of Chinese herbs,the present study was conducted to investigate the effects of different Chinese herbs against MS.Drug sensitivity test was carried out to check the efficacy of top five herbal extracts out of from 12 herbal extracts against the MS and on the bases of this experiment we got top five medicine.In this experiment,micro dilution method(minimum inhibitory concentration of mycoplasma cysts)was used by using aseptic snail test tubes and Oxford cup method was used for confirmation.The results showed that,the sensitivity of MS towards Chinese rhubarb and Coptis chinensis was the highest with Bacteriostatic range value of 15.89-16.98 mg/ml,followed by the Honeysuckle,Codonopsis pilosula,and Licorice having the Bacteriostatic range value of 25.86,27.67 and 33.48 mg/ml.The oxford cup bacteriostatic range of Chinese rhubarb and Coptis chinensis,Honeysuckle,Codonopsis pilosula,and Licorice was higher than 24 mm,with the diameter of 24 mm were expressed by S means sensitive,while from 14～24 mm diameter was expressed by I means,moderately sensitive and diameter below from the 14 mm was expressed by R which mean insensitive or resistant.After extraction of herbs,high performance liquid chromatography(HPLC)was performed for every Chinese herbal medicine to quantify herb ingredients.It was ensured that each Chinese herbal medicine contained components having Anti-bacterial properties and each mililiter of solution contained 1 g of plant material.After all processing,Chinese Rhubarb had antibacterial properties via chrysophanol,Gardenia jasminoides had antibacterial properties via Berberine,Honeysuckle had antibacterial properties via luteolin,Codonopsis pilosula had antibacterial properties via alkaloids and Nitrogen-containing components and Licorice had antibacterial properties glycyrrhizin.The fingerprint of samples from different origins had strong characteristics and specificity,and the stability of 10 common peaks was good.The average similarity of fingerprints of was between 0.0928 and 0.0983,which met the requirements of relevant departments of the state.The non-common peak area of each sample accounted for less than 10%of the total peak area,which met the requirements of fingerprint.In order to determine the toxicity of extracts of Chinese herbal medicine after compatibility according to traditional Chinese medicine theory and to verify the safety of the compound preparation of Chinese herbal medicine,acute toxicity test was used to determine the LD50 and its confidence interval in mice,so as to provide reliable basis for clinical safe and rational drug use.Mice(48,196 ICR)having 18-22g weight were randomly divided into 6 groups and 8 mice in each group.One group was the blank and served as a control group and other 5 groups were experimental groups.The experimental groups were injected with herbal extracts with the dosage rate of 10000,20000,40000,80000 and 160000 mg/kg body weight,respectively.According to the formula Slog LD50=d,the standard error Slog LD50=0.01067 was calculated,and the 95%confidence interval of LD50 was 38990.26-42482.05 mg/kg.(d is the logarithmic distance in formula,P is the mortality of each group,n is the number of animals in each group).In this experiment,the LD50 of the compound Chinese medicine preparation was 41438 mg/kg in mice,which was much more than 10000 mg/kg,so the compound Chinese medicine preparation was safe and non-toxic.To find out infection pathway for Mycoplasma Synoviae,12 chickens of six-week-old were divided into four groups,one group was injected with 0.5 ml saline as control,the other three groups were injected with 0.5 ml bacterial solution through respiratory tract,digestive tract and through skin for three consecutive days.Results showed that,after 18 days of infection,the joints of the respiratory tract group were swollen,with fluctuate feeling of hands,slow movement,severe limping,swelling around the eyelids,tears in eyes,mild symptoms in the respiratory group.The temperature of the respiratory tract group fluctuated greatly than other groups.The PCR results showed that Mycoplasma Synoviae was positive in respiratory tract group.No expected amplification fragments were found in other groups.The results of antibody detection showed that all the groups were positive with MS.According to the ELISA kit results determination,the respiratory tract group got infection after 9 days and skin infection group was showed positive signs after 12 days,and other groups was positive after 15 days.From this experiment we can conclude that respiratory tract is the fastest and easy way to catch infection for Mycoplasma synovaie in chicken.This experiment was carried out through respiratory tract infection of chicken with MS.MS bacteria(0.5 ml)were injected through the respiratory tract for three days.Total 60 chicken were used in six group(10chicken/group).Groups were named as,1,2,3,4,5,and 6.Group one was negative control,group 2 was positive control,group three was given antibiotics and group 4,5 and 6 was given herbal extracts@2ml/1,1.5ml/l and lml/l,respectively.After infection,the temperature of the chicken through vent was measured every day.The blood collection was done after every three days(at 3,6,9,12,15 and 18 d)after the infection,and the serum was separated and frozen at-80℃.The results of the antibody test of the six separated serum showed that group 3(antibiotic)and 4 which were supplemented with herbal extracts(2 ml/l)had better recovery response towards MS as compared to other treatments groups,the positive results were found in each group at day 6,while at day 9 and 12,there was no positive sign in group 3 but other groups(4,5 and 6)still showed positive response.At day 15,groups 3 and 4 showed negative response while group 5 and 6 still showed positive response.At day 18,all treatments groups(3,4,5)showed negative,except group 6,At day 18 all treatments groups(3,4,5 and 6)showed negative results towards MS.Based on findings it is concluded that Chinese herbs extract of Chinese rhubarb,Coptis chinensis,Honeysuckle,Codonopsis pilosula and Licorice supplementation in drinking water@2ml/bird/day for 3 days successfully recover mycoplasma synovia.Therefore,Chinese herbs extract of Chinese rhubarb,Coptis chinensis,Honeysuckle,Codonopsis pilosula and Licorice has potential to be alternative of antibiotic against mycoplasma synovia.Compound Chinese Herbal extracts effect on Immune organs weight and immune Performance of chicken were examined randomly divided into 5 treatments,there were 10 chickens in each treatment The result revealed that:Compound Chinese herbal medicine increased the weight and their immune indexes of thymus,spleen and bursa.The weight of spleen and its index of Chinese herb medicine group was higher than the antibiotics group and control group in the third week and the different was significantly.After 7 days of immunization,the immune response of herbal group treatment groups was higher than the control group,and antibiotics group and the difference was significantly.This kind of different would persist to the 21st day.Mean while the result of immune index and immune organs revealed that,compound Chinese herbal medicine increased the weight and their immune indexes of thymus,spleen and bursa,which have a vital role in immunity of birds.The results of ELISA antibodies titer of IgG,IgA,IgM and IgY,in all group are following:at the first week of supplementation of herbal extracts the value of IgG are highest in group 3 followed by group 4,2,1 and 5.Highest value of IgA were found in group 4,followed by group 5,1,2 and 3.while highest value of IgM and IgY in group 4 and 1 as compared to other group,respectively.At the fourth week of supplementation of herbal extracts the value of IgG is the highest in group 3 followed by group 2,4,5 and 1.Highest value of IgA were found in group 3,followed by group 2,4,4and 5.while highest value of IgM and IgY also observed in group 3 as compared to other group,respectively.At the end of trial,two birds from each replicate were randomly picked up,individually weighed and slaughtered for intestinal histomorphology analysis.Samples from distal portion of duodenum,jejunum and ileum of small intestine(without material)were collected from slaughtered birds and fixed in 10%buffered formalin saline.Results revealed that,the highest value of duodenal villus height,crypt depth and villus area was observed in treatment C(group 4)followed by B(group 3),D(group 5),E(group 6)and A(group1),respectively.Meanwhile,the highest values of jejunum and ileum villus height,crypt depth and villus area were observed in treatment C(group 4)followed by B(group 3),D(group 5),E(group 6),and A(group 1),respectively.To determine the haematology and digestibility 40 chicken were used in 4 groups(A,B,C and D),A was control group,B,C and D were given herbal extract@2ml/l,1.5 ml/l and 1 ml/l,respectively.The birds(4 birds/replicate)were slaughtered and blood samples(5 ml)were collected.For digestibility trays were placed under each pen for the feces collection.Feces were collected for forty-eight hours to measure nutrient digestibility.Results showed that addition of Chinese herbal extract into drinking water showed a significant(P<0.05)effect on blood glucose,cholesterol and erythrocytes(RBC).The study finding showed a significant(P<0.05)reduction of blood glucose and cholesterol in the birds of treatments B and C.However,red blood cells(RBC)count significantly(P<0.05)improved in the birds of treatment C.Whereas,leukocytes(WBC),hemoglobin,and packed cell volume stayed same due to the supplementation of these Chinese herbal extracts.The addition of Chinese compound herbal extracts exhibited a significant(P<0.05)influence on digestibility of CP(crude protein)in birds in all treatments in comparison to control treatment.The finding showed crude protein digestibility and crude lipids was significantly(P<0.05)higher in treatment B,while all other treatments including control group showed lower protein digestibility.While the minimum digestion of CF(crude fat)and CP(crude protein)was recorded in the control treatment(A),supplemented water without addition of leaf extract.Conversely,digestibility of CF persisted to be unaffected due to the addition of different HPLE,in all the treatments.Supplementation of compound Chinese herbal medicine exhibited better nutrient digestibility of crude protein and ether extract of the rations fed to the birds in treated groups coupled with improved intestinal histomorphology and hematological parameters studied.Therefore,use of compound Chinese herbal medicine chicken is recommended as inexpensive and efficient growth promoting agent without residual effects like antibiotic growth promoter. |
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