羊伪结核和大肠杆菌病病原分离鉴定及其二联灭活苗的研制
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摘要
羊大肠杆菌病是由致病性大肠杆菌引起的一种以腹泻或急性败血症为特征的羊
传染病。主要感染数日至 6 月龄的羔羊。以高热、呼吸困难、神经症状、胸膜肺炎和
急性死亡为特征的羔羊败血型大肠杆菌病致死率最高,给养羊业带来巨大的经济损
失。从具有典型症状的不同羊场患病羊病料中分离到 20 株革兰氏阴性不溶血短杆菌,
结合生化试验结果和病理变化,确定病原菌为大肠杆菌。经血清型鉴定,发现 20 株
分离菌中有 18 株血清型为O78。分离菌对青霉素、氨苄青霉素耐受,对其它药物敏
感程度存在较大差异。致病性试验和回归试验证实,分离菌可致实验动物和羊发病死
亡。从致死动物病料中分离到了与自然病例相一致的大肠杆菌。
羊的另外一种由伪结核棒状杆菌引起的人畜共患慢性传染病—羊伪结核(羊干酪
性淋巴结炎)的发病率近年来在很多羊场呈逐年上升趋势。该病临床症状不明显,致
死率低,常被人们忽视。其主要特征是在患羊体表淋巴结或某些实质性脏器形成干酪
样脓肿,使患羊消瘦,生产性能低下或死胎,是国际公认的难以防治和彻底消除的传
染病之一。对可疑病羊病料进行病原菌分离鉴定,发现一种无荚膜、无芽孢的革兰氏
阳性多形性球杆菌,对青霉素和利福平等多种药物耐受,对实验动物有一定致病性和
致死性,可复制出与原始病例相一致的动物模型。结合生化试验特点和血清学试验结
果、临床症状等,判定其为伪结核棒状杆菌。
由于药物治疗不但易产生耐药菌株,而且需要大量人力、财力和物力,因此我们
尝试用分离的地方菌株制作联苗对羊的这两种传染病进行预防。以上述鉴定过的羊大
肠杆菌和伪结核棒状杆菌作为制苗菌种接种适宜培养基进行大量增殖,经甲醛灭活后
按一定比例加入佐剂,制成羊伪结核及大肠杆菌病多价二联油乳灭活苗,每毫升疫苗
含两种菌均在 100 亿左右。多项实验室检查证明,该二联苗稳定性好,使用安全,无
不良反应。实验动物免疫后 28 天,经攻毒试验及对血液中淋巴细胞比率和抗体的检
测,发现该疫苗免疫原性好,对实验动物的保护率为 100%。杨凌某羊场 200 多只羔
羊免疫接种该二联苗后,4 个月来未发现一例可疑病例,有效预防和抑制了羊大肠杆
菌病及伪结核病的发生和流行,效果可靠,可进一步推广应用。
Colibacillosis in sheep and goat is an infection disease with diarrhea or
acute septicemia caused by pathogenic E.coli. The lambs of days to six
months old are infected most easily. The mortality of septicemia typal
Colibacillosis in lambs is very high, which characters is high fever, labored
breathing, nerves signs, pleura-pneumonia and acute death. The disease brings
about huge loss to income of sheep and goat-raising. The twenty strains of
short bacillus were isolated died lambs with typical clinical in various plants.
The isolates all are gram negative and not hemolysis. Combinining the results
of biochemical test and pathology signs together, it is indicated that the
pathogenic bacterium is E.coli. Serological reaction test indentified that the
serotypes of eighteen strains are O78. The isolates are resistance to penicillin
and ampicillin, the different of sensibility to other drugs is very vast. Animal
infection test prove that the isolates can cause animal fall disease or death,
and E.coli. same as nature case can be isolated from dead body of these
animals.
In recent years, Pseudotuberculosis in sheep and goat (Caseous
Lymphadenitis) is climbing up continually. It is the chronic zoonotic
infectious disease caused by Corynebacterium pseudotuberculosis. This
infection is often neglected by people because of its unnoticed clinical
symptom and low mortality. It can cause some lymph nodes or organs caseous
abscesses and bring about emaciation, poor doers, fetus die in uterus to sheep
or goat. So, the disease is one of the infections to control and remove in the
world. The same ball-bacillus was isolated from cervix lymphatic abscess of
diseased goats in some plants. It is gram positive without capsule and spore,
resistant to Penicillin G and Rifampin etc.. It can cause experiment animals
fall in diseases or death, and can copy the same animal models as nature case
on goats. Combining results of biochemical test, serology test and clinical
signs, it is concluded that the pathogenicity is Corynebacterium
pseudotuberculosis.
In order to prevent and control colibacillosis and pseudotuberculosis, we
tried to make the bi-combined oil inactivated vaccine from typical isolated
bacterium strains. After the isolated E.coli and Corynebacterium
pseudotuberculosis were cultured on suitable nutrition to proliferate and
Abstract Ⅲ
inactivated by Formaldehyde ,then add into adjuvant in proportion and make
into the mutti-valent bi-combined oil inactivated vaccine of the two species
bacteria. There are 10 billions E.coli CFU and Corynebacterium
pseudotuberculosis CFU in every milliliter vaccine. Laboratory tests indicate
that the vaccine is stable, effective and safe. Examining the lymphocyte
percentages in WBC of goats in 28 days after inoculation and through
infection test, the vaccine is proven that it have fine immunogenicity and the
rate of protection to all experiment animals is 100%. After the vaccine is
injected into about two hundred lambs of a plant in Yangling, one doubtful
case have never been found in later 4 months. This bi-combined oil
inactivated vaccine protected Colibacillosis of sheep and goat to outbreak and
spread, also inhibited pseudotuberculosis as well. So the vaccine should be
improved and utilized extensively.
传染病。主要感染数日至 6 月龄的羔羊。以高热、呼吸困难、神经症状、胸膜肺炎和
急性死亡为特征的羔羊败血型大肠杆菌病致死率最高,给养羊业带来巨大的经济损
失。从具有典型症状的不同羊场患病羊病料中分离到 20 株革兰氏阴性不溶血短杆菌,
结合生化试验结果和病理变化,确定病原菌为大肠杆菌。经血清型鉴定,发现 20 株
分离菌中有 18 株血清型为O78。分离菌对青霉素、氨苄青霉素耐受,对其它药物敏
感程度存在较大差异。致病性试验和回归试验证实,分离菌可致实验动物和羊发病死
亡。从致死动物病料中分离到了与自然病例相一致的大肠杆菌。
羊的另外一种由伪结核棒状杆菌引起的人畜共患慢性传染病—羊伪结核(羊干酪
性淋巴结炎)的发病率近年来在很多羊场呈逐年上升趋势。该病临床症状不明显,致
死率低,常被人们忽视。其主要特征是在患羊体表淋巴结或某些实质性脏器形成干酪
样脓肿,使患羊消瘦,生产性能低下或死胎,是国际公认的难以防治和彻底消除的传
染病之一。对可疑病羊病料进行病原菌分离鉴定,发现一种无荚膜、无芽孢的革兰氏
阳性多形性球杆菌,对青霉素和利福平等多种药物耐受,对实验动物有一定致病性和
致死性,可复制出与原始病例相一致的动物模型。结合生化试验特点和血清学试验结
果、临床症状等,判定其为伪结核棒状杆菌。
由于药物治疗不但易产生耐药菌株,而且需要大量人力、财力和物力,因此我们
尝试用分离的地方菌株制作联苗对羊的这两种传染病进行预防。以上述鉴定过的羊大
肠杆菌和伪结核棒状杆菌作为制苗菌种接种适宜培养基进行大量增殖,经甲醛灭活后
按一定比例加入佐剂,制成羊伪结核及大肠杆菌病多价二联油乳灭活苗,每毫升疫苗
含两种菌均在 100 亿左右。多项实验室检查证明,该二联苗稳定性好,使用安全,无
不良反应。实验动物免疫后 28 天,经攻毒试验及对血液中淋巴细胞比率和抗体的检
测,发现该疫苗免疫原性好,对实验动物的保护率为 100%。杨凌某羊场 200 多只羔
羊免疫接种该二联苗后,4 个月来未发现一例可疑病例,有效预防和抑制了羊大肠杆
菌病及伪结核病的发生和流行,效果可靠,可进一步推广应用。
Colibacillosis in sheep and goat is an infection disease with diarrhea or
acute septicemia caused by pathogenic E.coli. The lambs of days to six
months old are infected most easily. The mortality of septicemia typal
Colibacillosis in lambs is very high, which characters is high fever, labored
breathing, nerves signs, pleura-pneumonia and acute death. The disease brings
about huge loss to income of sheep and goat-raising. The twenty strains of
short bacillus were isolated died lambs with typical clinical in various plants.
The isolates all are gram negative and not hemolysis. Combinining the results
of biochemical test and pathology signs together, it is indicated that the
pathogenic bacterium is E.coli. Serological reaction test indentified that the
serotypes of eighteen strains are O78. The isolates are resistance to penicillin
and ampicillin, the different of sensibility to other drugs is very vast. Animal
infection test prove that the isolates can cause animal fall disease or death,
and E.coli. same as nature case can be isolated from dead body of these
animals.
In recent years, Pseudotuberculosis in sheep and goat (Caseous
Lymphadenitis) is climbing up continually. It is the chronic zoonotic
infectious disease caused by Corynebacterium pseudotuberculosis. This
infection is often neglected by people because of its unnoticed clinical
symptom and low mortality. It can cause some lymph nodes or organs caseous
abscesses and bring about emaciation, poor doers, fetus die in uterus to sheep
or goat. So, the disease is one of the infections to control and remove in the
world. The same ball-bacillus was isolated from cervix lymphatic abscess of
diseased goats in some plants. It is gram positive without capsule and spore,
resistant to Penicillin G and Rifampin etc.. It can cause experiment animals
fall in diseases or death, and can copy the same animal models as nature case
on goats. Combining results of biochemical test, serology test and clinical
signs, it is concluded that the pathogenicity is Corynebacterium
pseudotuberculosis.
In order to prevent and control colibacillosis and pseudotuberculosis, we
tried to make the bi-combined oil inactivated vaccine from typical isolated
bacterium strains. After the isolated E.coli and Corynebacterium
pseudotuberculosis were cultured on suitable nutrition to proliferate and
Abstract Ⅲ
inactivated by Formaldehyde ,then add into adjuvant in proportion and make
into the mutti-valent bi-combined oil inactivated vaccine of the two species
bacteria. There are 10 billions E.coli CFU and Corynebacterium
pseudotuberculosis CFU in every milliliter vaccine. Laboratory tests indicate
that the vaccine is stable, effective and safe. Examining the lymphocyte
percentages in WBC of goats in 28 days after inoculation and through
infection test, the vaccine is proven that it have fine immunogenicity and the
rate of protection to all experiment animals is 100%. After the vaccine is
injected into about two hundred lambs of a plant in Yangling, one doubtful
case have never been found in later 4 months. This bi-combined oil
inactivated vaccine protected Colibacillosis of sheep and goat to outbreak and
spread, also inhibited pseudotuberculosis as well. So the vaccine should be
improved and utilized extensively.
引文