| TB is a kind of zoonoses caused by Mycobacterium tuberculosis, it was reported that in theremote areas where living and health conditions are more poorer, while in these areas, people,cattle, deer and other susceptible animals are more easyer contact with each other, which is also a greater risk of TB transmission, so in rural areas and in the dairy, deer farm and so on TBprevention and control work carried out, so only focusing on the prevention and treatment ofsusceptible animals is not enough, as the treatment measures of eliminating infection sources,preventing the infection media and protecting animals susceptible to infectious disease, it demandsthat we have to pay attention to the animals infected with Mycobacterium tuberculosis, in order todevelop a reasonable livestock tuberculosis program in conjunction with one joint implementationof TB prevention and control plan to human.To this aid, in this trial we collected suspected cow, deer's lymph nodes through Changchunarea. After Shreddering, grinding, digestion, we inoculated the samples to general Roche medium,selective 7H9 liquid medium, sodium pyruvate medium isolation and culturing in 37 , afte℃robtaining the feature colonies, the acid-fast staining, TCH and PNB identification culture and PCRmethod based on hupB gene are carried to identify bacteria type, human or bovine type, throughwhich we can probably understand the tuberculosis prevalence of cattle,deer or other animals inthe rural areas and farms. As a result, we got 100 cases of isolated mycobacteria from the 400collected samples from the lymph nodes, of which includes 90 non-tuberculosis mycobacteriumcases and 10 Mycobacterium tuberculosis cases including 2 human type and 8 bovisMycobacterium, human-type cases up to 20%. It is reported that in human clinical cattle-typeaccounts for about 6%. After the drug sensitivity test of the first-line anti-TB drugs for the clinicalbacteria, 61% mycobacteria are single-drug or multi-drug resistant. From the preliminary results,we know that TB not only an economic issue, but also a serious social problem. High degree ofattention should be aroused to TB prevention and control for livestock.The characteristic of Mycobacterium tuberculosis that it can evade the the attack of humanimmune system, which giving more difficult to prevent and control MT; and the surge of resistant, especially multi-drug resistant MT, which makes the use of the one and the second-line anti-TB medicine more and more difficult. So infected with multidrug-resistant TB tend to keep persistent deterioration of the disease and eventually become fatal, so the research of anti-TB drugs to fight TB is already a priority, since traditional Chinese herbal medicine has been used in anti-TB treatment since ancient times. Therefore, in the absence of new vaccines and drugs, screening an effective Chinese herbal medicine has an important meaning. A few of Chinese herbal monomer compounds for the standard strains of Mycobacterium tuberculosis ATCC 27294 H37Rv and the animal isolated clinical strains, by using the 96-hole plate microdilution susceptibility test Methods (MABA method), from which we got oleanolic acid with a lower MIC value, and using the 96-well micro-plate ot test the joint microdilution test method (FIC) of oleanolic acid with several commonly used first-line anti-TB drugs. When against 27294 the FICI of the combined results of oleanolic acid with rifampicin, isoniazid, streptomycin and ethambutol were 0.531, 1.062, 0.530 and 0.531 respectivelly, exceptible for INH, combined with the RFP, SM and EMB all shown a good synergistic effect; while against the clinical isolates, the joint effect whether targeted at the drug-resistant or susceptible strains of clinical, FICI are much better than 27294. For the drug-resistant clinical isolates of M. tuberculosis, the FICI extent of OA combined with INH, RFP and EMB are 0.121～0.347, 0.113～0.168 and 0.093～0.266 respectivelly. MTT cytotoxicity assay showed that, the maximum non-cytotoxic concentration of OA against the African green monkey kidney cells (vero cells) is 20μg/mL, while inhibition of the growth of the concentration of OA against the strain ATCC 27294 is 16μg/mL, which means that oleanolic acid is not cytotoxic on human cells. So OA is a potential anti-TB drugs.In summary, animal disease pathogens Mycobacterium tuberculosis, is not only bovine type, but also some Mycobacterium tuberculosis type, that is not only bovine type transmission among the animals, may also be due to human and illness in close contact with livestock or human consumption without strict disinfection and sterilization of the diseased livestock foods and animals infected with TB incidence of bovine or human type. When treatment with the incidence of contact with animals infected with TB patients, to the same time, incidence of potentially contaminated animals and animal populations to synchronize the monitoring, prevention and control measures such as immunization and quarantine. As the traditional one and second-line anti-tuberculosis drugs have been used for several decades, natural strains of TB epidemic in part because a large amount of a long-term, second-line anti-tuberculosis drugs, selective stimulation, has been divided many single-drug resistance, multi-drug resistance and even is a super-drug-resistant Mycobacterium tuberculosis, resulting in clinically selected from a limited anti-TB drugs are very difficult, when, and if we failure to provide an alternative and effective drug, will make the drug-resistant tuberculosis is more rampant, making matters of TB prevention and control work more worse . Therefore, the pressing task is to find effects of clear and less toxic anti-TB drugs substitutability as soon as possible. To this aid, the domestic anti-TB China's traditional herbal medicine into the people's attention, want to filter out from the exact effect of Mycobacterium tuberculosis, monomer componant are not toxic to animals, or the body can withstand the toxicity of the smaller monomer compounds, and guide the next step anti-branching Bacillus drug development and drug target against the compounds were the subject of synthetic anti-tuberculosis drug research. |
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