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南寧某奶牛場(chǎng)乳房炎監(jiān)測(cè)與病原調(diào)查

發(fā)布時(shí)間:2018-07-14 07:29
【摘要】:奶牛乳房炎(Bovine mastitis)在奶牛養(yǎng)殖業(yè)中是多見(jiàn)的疾病之一,它造成奶產(chǎn)量的減產(chǎn)以及低劣的原料乳,常常引發(fā)一些公共衛(wèi)生學(xué)問(wèn)題與食品安全問(wèn)題,奶牛乳房炎造成的危害,對(duì)于奶牛養(yǎng)殖戶,乳品加工行業(yè)來(lái)說(shuō)都是一個(gè)非常沉重的經(jīng)濟(jì)壓力。本次實(shí)驗(yàn)的奶樣都來(lái)至于南寧某奶牛場(chǎng),樣品先進(jìn)行乳汁質(zhì)量的檢測(cè);再進(jìn)行奶牛乳房炎主要病原菌分離鑒定及藥敏試驗(yàn);使用PCR檢測(cè)金黃色葡萄球菌的毒力基因和耐藥基因。具體研究如下:試驗(yàn)一:乳房炎與乳汁質(zhì)量的動(dòng)態(tài)監(jiān)測(cè)在2014年10月至2016年10月間,每月(個(gè)別月份缺失)定期隨機(jī)采集奶樣,應(yīng)用荷蘭產(chǎn)DEITA牌Somascope SmartTM體細(xì)胞檢測(cè)儀對(duì)奶樣進(jìn)行乳汁體細(xì)胞計(jì)數(shù),按不同SCC分類,SCC在50~100萬(wàn)/ml為隱性乳房炎。SCC50萬(wàn)/ml為正常乳汁,SCC數(shù)量越少,提示牛奶質(zhì)量越好。統(tǒng)計(jì)分析不同時(shí)期奶牛乳房炎的發(fā)生情況以及牛奶質(zhì)量情況。結(jié)果表明:不同泌乳月份奶牛個(gè)體SCC變化不同,泌乳月份奶牛個(gè)體乳SCC呈波動(dòng)性變化?傮w上來(lái)看,不同泌乳月份奶牛體乳SCC變化不同,泌乳月份奶牛體乳SCC呈波動(dòng)性變化,奶牛夏季體乳SCC呈波動(dòng)性變化最為明顯,冬季體乳SCC呈波動(dòng)性最小。在這20個(gè)月中,奶質(zhì)逐漸變好,其中2016年的奶質(zhì)是最好的。夏季、秋季的體細(xì)胞數(shù)比春季、冬季的高。合格奶樣整體呈現(xiàn)上升的走勢(shì),不合格奶樣整體也是呈現(xiàn)下降的走勢(shì)。試驗(yàn)二:奶牛乳房炎主要病原菌分離鑒定與藥敏試驗(yàn)通過(guò)細(xì)菌分離鑒定與生化鑒定,獲156株可疑菌株,生化鑒定確定出金黃色葡萄球菌57株,占36.5%、無(wú)乳鏈球菌13株、停乳鏈球菌7株、乳房鏈球菌6株,鏈球菌占16.7%、大腸桿菌73株,占46.8%。其中金葡菌的耐藥率是33.7%、鏈球菌的耐藥率是31.1%、大腸桿菌的的耐藥率是35.2%。鏈霉素、青霉素、四環(huán)素對(duì)大腸桿菌已經(jīng)100%失去作用;鏈霉素、青霉素也對(duì)金黃色葡萄球菌已經(jīng)是100%失去作用,恩諾沙星有較高的耐藥性,而對(duì)氨芐西林、羧芐西林、多西環(huán)素產(chǎn)生部分不敏感;鏈球菌對(duì)氨芐西林、鏈霉素、青霉素、紅霉素已經(jīng)100%耐藥,但三類菌對(duì)丁胺卡那、羧芐西林、頭孢哌酮、頭孢曲松、氧氟沙星都具有較高敏感性。試驗(yàn)三:金黃色葡萄球菌毒力基因檢測(cè)對(duì)上述生化鑒定的57株黃色葡萄球菌,再利用PCR檢測(cè)它的nuc、h1a、h1b、sea、seb毒力基因,nuc基因僅為金黃色葡萄球菌所具有而且具有高度保守性。結(jié)果表明:攜帶nuc基因的有57株(100%,)攜帶H1a基因的有13株(22.8%),攜帶H1b基因的有15株(23.6%)。同時(shí)攜帶有兩種的占7.0%。攜帶sea基因的有12株(21.1%),攜帶seb基因的有8株(14.0%),同時(shí)攜帶有兩種基因的占3.5%。在食品安全問(wèn)題上,仍需要重視。試驗(yàn)四:金黃色葡萄球菌耐藥基因檢測(cè)金黃色葡萄球菌是引起奶牛乳房炎最主要的病原菌之一,由于其感染率高,且呈現(xiàn)多重耐藥性。通過(guò)在線軟件設(shè)計(jì)金黃色葡萄球菌的紅霉素耐藥基因ermB、ermC和耐甲氧西林金黃色葡萄球mecA基因的引物,使用PCR檢測(cè),ermB、ermC毒力基因,結(jié)果表明:攜帶ermB基因的有10株(17.5%),ermC基因的有16株(28.1%),57株金黃色葡萄球菌基因的PCR檢測(cè),未檢測(cè)到攜帶mecA基因?茖W(xué)合理的使用抗生素,盡量避免耐藥株的產(chǎn)生。
[Abstract]:The dairy cow mastitis (Bovine mastitis) is one of the most common diseases in the dairy farming industry. It causes the reduction of milk production and the poor raw milk. It often causes some public health problems and food safety problems, and the harm caused by dairy cow mastitis. It is a very heavy classics for dairy farmers and dairy industry. The milk sample of this experiment came to a dairy farm in Nanning, the samples were first tested for milk quality, and the main pathogenic bacteria of Dairy Mastitis were separated and identified and the drug sensitivity test was carried out. PCR was used to detect the virulence genes and resistance genes of Staphylococcus aureus. Milk samples were collected regularly from October 2014 to October 2016. The milk samples were counted with the DEITA Somascope SmartTM somatic cell detector produced in Holland. According to different SCC classification, SCC was 50~100 million /ml as recessive mastitis.SCC50 million /ml as normal milk, and the less SCC, indicating milk quality. The quality of dairy cow mastitis and milk quality in different period of milking were analyzed. The results showed that the individual SCC changes of dairy cows in different lactation months were different, and the individual milk SCC of dairy cows fluctuated in month of lactation. On the whole, the SCC changes of dairy cows in different lactation months were different, and the milk SCC of dairy cows was fluctuant in lactating month. During the summer, milk quality was the least. In the 20 months, milk quality became better. In the 20 months, milk quality became better. In 2016, the milk quality was the best. In summer, the number of somatic cells in autumn was higher than that in spring and winter. Trend. Test two: the main pathogen isolation and identification and drug sensitivity test of dairy cow mastitis, 156 strains of Staphylococcus aureus, 13 strains of Streptococcus Lactococcus, 7 Streptococcus, 6 Streptococcus, 16.7% Streptococcus, 73 strains of Escherichia coli, and 73 strains of Escherichia coli, were identified by bacterial isolation and biochemical identification, 57 strains of Staphylococcus aureus, 13 Streptococcus, 7 Streptococcus, 6 strains of Streptococcus, 16.7% Streptococcus, 73 strains of Escherichia coli, 46.8%. The resistance rate of Staphylococcus aureus was 33.7%, the drug resistance rate of Streptococcus was 31.1%, the resistance rate of Escherichia coli was 35.2%. streptomycin, penicillin, and tetracycline had lost 100% effect on Escherichia coli; streptomycin, penicillin had also lost 100% to Staphylococcus aureus, and enrofloxacin was highly resistant to ampicillin and carboxyl. In benzicillin, doxycycline produced partial insensitivity; Streptococcus has 100% resistance to ampicillin, streptomycin, penicillin and erythromycin, but three types of bacteria have high sensitivity to amikacin, cicillin, cefoperazone, ceftriaxone and ofloxacin. Test three: 57 strains of Staphylococcus aureus virulence gene test for the above biochemical identification Staphylococcus aureus, and then using PCR to detect its NUC, H1a, H1B, sea, SEB virulence genes, the NUC gene is only with Staphylococcus aureus and highly conserved. The results show that 57 (100%) carrying the H1a gene carrying NUC (22.8%), and 15 (23.6%) carrying H1b genes, and two kinds of 7.0%. carrying the gene. 12 (21.1%) with sea gene, 8 (14%) carrying SEB gene and two genes in the food safety problem, still need to be paid attention to. Test four: Staphylococcus aureus resistance gene detection of Staphylococcus aureus is one of the most important pathogens causing cow mastitis, because of its high infection rate and more than one. Heavy drug resistance. Primers for the erythromycin resistant gene ermB, ermC and methicillin resistant Staphylococcus aureus mecA gene were designed by online software, and PCR detection, ermB, ermC virulence genes were used. The results showed that 10 (17.5%), 16 (28.1%) of ermC gene and 57 strains of Staphylococcus aureus gene were in PC, and 57 strains of Staphylococcus aureus. R detection showed that mecA gene was not detected. Scientific and rational use of antibiotics should be avoided as far as possible.
【學(xué)位授予單位】:廣西大學(xué)
【學(xué)位級(jí)別】:碩士
【學(xué)位授予年份】:2017
【分類號(hào)】:S858.23

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