哈尔滨部分奶牛场隐性乳房炎流行病学调查及中药治疗研究
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摘要
近年来,黑龙江省已成为我国最大的奶牛养殖基地之一,奶牛养殖规模发展迅猛,但是由于饲养管理水平相对较低,奶牛隐性乳房炎发病率居高不下,成为制约我省奶牛业发展的重要因素之一。本课题旨在研究哈尔滨地区奶牛隐性乳房炎的流行病学,感染性致病菌种类,筛选敏感性中药组方,制成中草药透皮剂并检测临床效果,为防治奶牛隐性乳房炎提供一定的技术支持。
本研究以哈尔滨地区部分奶牛场眼观泌乳期健康的奶牛为试验动物,采用流行病学调查法,乳汁体细胞计数法、CMT法、隐性乳房炎试纸法检测奶牛隐性乳房炎发病率,分析其流行情况;对患有隐性乳房炎的乳汁进行体外细菌培养、分离鉴定,应用中药体外抑菌试验、杀菌试验,随机区组确定奶牛隐性乳房炎致病菌敏感性中药组方,开发隐性乳房炎治疗性透皮剂。结果发现:
1.哈尔滨地区奶牛隐性乳房炎阳性率为23.68%(398/1681),乳区平均阳性率为7.21%;奶牛隐性乳房炎发病率在低温季节及季节转变期、泌乳后期(201~305d)、砖面运动场上发病率较高,并随着奶牛年龄及胎次增加而增高。
2.对50份患有隐性乳房炎的奶样进行细菌的分离,细菌分离率达到96%(48/50)。共分离出6种共计66株细菌,其中金黄色葡萄球菌12株,表皮葡萄球菌11株,腐生葡萄球菌4株,停乳链球菌11株,无乳链球菌6株,乳房链球菌5株,大肠埃希菌9株,棒状杆菌2株,沙门氏菌4株,蜡状芽孢杆菌2株。其中样品中只分离出1种细菌的占90.91%(60/66),样品中分离出2种以上的占9.09%(6/66)。所鉴定的主要致病菌种类为金黄色葡萄球菌、表皮葡萄球菌、停乳链球菌、大肠埃希菌,这四种细菌占总分离细菌的65.15%(43/66)。
3.上述检出的病原菌对头孢曲松、恩诺沙星、氧氟沙星、环丙沙星比较敏感,对其他抗菌素敏感性比较低,并产生了不同程度的耐药性。应用7个中药组方对主要致病菌金黄色葡萄球菌、停乳链球菌、大肠埃希菌采取体外抑菌、杀菌试验及随机区组试验,得到复方Ⅱ:黄连、红花、连翘、黄芩组合为最佳组方。采用东北农业大学外科教研室自行研制的透皮剂制作方法制取中药透皮剂,并在奶牛场选取试验奶牛采取试验,结果表明:中药透皮剂组与氧氟沙星组临床效果基本一致(P>0.05),为哈尔滨地区开发天然、绿色、无药物残留的防治性药剂奠定了技术基础。
In recent years, Heilongjiang province has become one of dairy cow bases. Dairy scale is rapid development, but the level of management is relatively low; Subclinical Mastitis of dairy cow that is high morbidity is one of the most important factors which prohibit the development of dairy industry. In order to provide technical support to prevent Subclinical Mastitis of dairy cow , this research in Harbin investigate its epidemiological conditions , types of transmissibility pathogenic bacteria , screening sensitive traditional Chinese medicine formula and preparation and inspection transdermal preparation .
The experimental animals are the lactation and healthy cows of a part of dairy farms in Harbin. Main methods are as follow : epidemiological experiment , lacto-somatic cell counting , CMT , test paper detecting disease rate ; Milk of Subclinical Mastitis cows implement a series of tests , including cultivating , isolating and identify bacteria in vitro ; antibacterial , bactericidal test of the traditional Chinese medicine in vitro ; we will do randomized block experiment for knowing about pathogenic bacteria sensitive traditional Chinese medicine formula and developing transdermal preparation . The results showed that:
1. In Harbin, positive rate of Subclinical Mastitis is 23.68% (398/1681), the average positive rate of breast area is 7.21%; the incidence of Subclinical Mastitis is higher when cows are in the seasonal changes, in the low temperature season, the period of the late lactation (201 ~ 305d) and live in the brick stadium, and shows positive correlation with age and labour times.
2. Isolate from 50 pairs of bacteria samples of milk suffered subclinical mastitis. The rate of bacterial isolation reached 96% (48/50). We totally got six kinds and a total of 66 isolated strains of bacteria, of which 12 strains of Staphylococcus aureus, Staphylococcus epidermidis 11 strains, S.saprophyticus 4 strains, Streptococcus dysgalactiae 11 strains, Streptococcus agalactiae 6 stains, Streptococcus uberis 5 strains, Escherichia coli 9 strains, 2 strains of Corynebacterium, Salmonella 4 strains, 2 strains of Bacillus cereus. Samples in which only isolated a bacterium accounted for 90.91% (60/66), samples isolated from two or more accounted for 9.09% (6 / 66). The main types of pathogens are Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus dysgalactiae, Escherichia coli, these four bacteria account for 65.15% (43/66) among the total separation of bacteria.
3. Detection pathogen is sensitive to ceftriaxone, enrofloxacin, ofloxacin; ciprofloxacin. The sensitivity to other drugs is relatively low and showed different degrees of drug resistance. Through antibacterial, bactericidal test in vitro and randomized block experiment to the major pathogens (Staphylococcus aureus, Streptococcus dysgalactiae, Escherichia coli) with seven traditional Chinese medicine formulas, we obtained compoundⅡ: coptis chinensis, safflower, Forsythia, baicalin combination is the best group of parties. Apply production methods of transdermal preparation which is self-developed by Northeast Agricultural University, Surgery teaching and research to produce traditional Chinese medicine transdermal agent, and selected cows in dairy to start test. Results showed that: the group of traditional Chinese medicine transdermal agent and the group of ofloxacin showed similar clinical effects (P >0.05), so as to lay a solid foundation to make this square transdermal agent the best one to control Harbin Subclinical mastitis.
本研究以哈尔滨地区部分奶牛场眼观泌乳期健康的奶牛为试验动物,采用流行病学调查法,乳汁体细胞计数法、CMT法、隐性乳房炎试纸法检测奶牛隐性乳房炎发病率,分析其流行情况;对患有隐性乳房炎的乳汁进行体外细菌培养、分离鉴定,应用中药体外抑菌试验、杀菌试验,随机区组确定奶牛隐性乳房炎致病菌敏感性中药组方,开发隐性乳房炎治疗性透皮剂。结果发现:
1.哈尔滨地区奶牛隐性乳房炎阳性率为23.68%(398/1681),乳区平均阳性率为7.21%;奶牛隐性乳房炎发病率在低温季节及季节转变期、泌乳后期(201~305d)、砖面运动场上发病率较高,并随着奶牛年龄及胎次增加而增高。
2.对50份患有隐性乳房炎的奶样进行细菌的分离,细菌分离率达到96%(48/50)。共分离出6种共计66株细菌,其中金黄色葡萄球菌12株,表皮葡萄球菌11株,腐生葡萄球菌4株,停乳链球菌11株,无乳链球菌6株,乳房链球菌5株,大肠埃希菌9株,棒状杆菌2株,沙门氏菌4株,蜡状芽孢杆菌2株。其中样品中只分离出1种细菌的占90.91%(60/66),样品中分离出2种以上的占9.09%(6/66)。所鉴定的主要致病菌种类为金黄色葡萄球菌、表皮葡萄球菌、停乳链球菌、大肠埃希菌,这四种细菌占总分离细菌的65.15%(43/66)。
3.上述检出的病原菌对头孢曲松、恩诺沙星、氧氟沙星、环丙沙星比较敏感,对其他抗菌素敏感性比较低,并产生了不同程度的耐药性。应用7个中药组方对主要致病菌金黄色葡萄球菌、停乳链球菌、大肠埃希菌采取体外抑菌、杀菌试验及随机区组试验,得到复方Ⅱ:黄连、红花、连翘、黄芩组合为最佳组方。采用东北农业大学外科教研室自行研制的透皮剂制作方法制取中药透皮剂,并在奶牛场选取试验奶牛采取试验,结果表明:中药透皮剂组与氧氟沙星组临床效果基本一致(P>0.05),为哈尔滨地区开发天然、绿色、无药物残留的防治性药剂奠定了技术基础。
In recent years, Heilongjiang province has become one of dairy cow bases. Dairy scale is rapid development, but the level of management is relatively low; Subclinical Mastitis of dairy cow that is high morbidity is one of the most important factors which prohibit the development of dairy industry. In order to provide technical support to prevent Subclinical Mastitis of dairy cow , this research in Harbin investigate its epidemiological conditions , types of transmissibility pathogenic bacteria , screening sensitive traditional Chinese medicine formula and preparation and inspection transdermal preparation .
The experimental animals are the lactation and healthy cows of a part of dairy farms in Harbin. Main methods are as follow : epidemiological experiment , lacto-somatic cell counting , CMT , test paper detecting disease rate ; Milk of Subclinical Mastitis cows implement a series of tests , including cultivating , isolating and identify bacteria in vitro ; antibacterial , bactericidal test of the traditional Chinese medicine in vitro ; we will do randomized block experiment for knowing about pathogenic bacteria sensitive traditional Chinese medicine formula and developing transdermal preparation . The results showed that:
1. In Harbin, positive rate of Subclinical Mastitis is 23.68% (398/1681), the average positive rate of breast area is 7.21%; the incidence of Subclinical Mastitis is higher when cows are in the seasonal changes, in the low temperature season, the period of the late lactation (201 ~ 305d) and live in the brick stadium, and shows positive correlation with age and labour times.
2. Isolate from 50 pairs of bacteria samples of milk suffered subclinical mastitis. The rate of bacterial isolation reached 96% (48/50). We totally got six kinds and a total of 66 isolated strains of bacteria, of which 12 strains of Staphylococcus aureus, Staphylococcus epidermidis 11 strains, S.saprophyticus 4 strains, Streptococcus dysgalactiae 11 strains, Streptococcus agalactiae 6 stains, Streptococcus uberis 5 strains, Escherichia coli 9 strains, 2 strains of Corynebacterium, Salmonella 4 strains, 2 strains of Bacillus cereus. Samples in which only isolated a bacterium accounted for 90.91% (60/66), samples isolated from two or more accounted for 9.09% (6 / 66). The main types of pathogens are Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus dysgalactiae, Escherichia coli, these four bacteria account for 65.15% (43/66) among the total separation of bacteria.
3. Detection pathogen is sensitive to ceftriaxone, enrofloxacin, ofloxacin; ciprofloxacin. The sensitivity to other drugs is relatively low and showed different degrees of drug resistance. Through antibacterial, bactericidal test in vitro and randomized block experiment to the major pathogens (Staphylococcus aureus, Streptococcus dysgalactiae, Escherichia coli) with seven traditional Chinese medicine formulas, we obtained compoundⅡ: coptis chinensis, safflower, Forsythia, baicalin combination is the best group of parties. Apply production methods of transdermal preparation which is self-developed by Northeast Agricultural University, Surgery teaching and research to produce traditional Chinese medicine transdermal agent, and selected cows in dairy to start test. Results showed that: the group of traditional Chinese medicine transdermal agent and the group of ofloxacin showed similar clinical effects (P >0.05), so as to lay a solid foundation to make this square transdermal agent the best one to control Harbin Subclinical mastitis.
引文
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