本文選題：豬源鏈球菌 + 流行病學(xué)�。� 參考：《河南科技大學(xué)》2017年碩士論文

【摘要】：近年來,豬鏈球菌病在我國廣泛流行并致病,已成為嚴(yán)重危害養(yǎng)豬業(yè)發(fā)展的主要病原菌之一。本研究對豬源鏈球菌的病原流行病學(xué)、毒力特性、生物學(xué)特性等進(jìn)行了系統(tǒng)研究,得到了以下研究結(jié)果:1.通過對豬源鏈球菌的分離鑒定(2014.09~2015.12),并對前期實(shí)驗(yàn)室分離鑒定結(jié)果進(jìn)行統(tǒng)計(jì)分析(2011.01~2014.08),結(jié)果表明,2011~2015年間,從河南及其周邊地區(qū)規(guī)模養(yǎng)殖場和部分散養(yǎng)農(nóng)戶2 521份發(fā)病豬的肺臟等病料組織中分離鑒定出772株鏈球菌(Streptococcus),總分離率為30.6%。通過基于gdh基因的特異性PCR方法對其中310株豬源鏈球菌進(jìn)行了PCR鑒定,共鑒定出135株豬鏈球菌(S.suis),占43.5%。通過基于cps基因的特異性PCR方法對135株豬鏈球菌進(jìn)行了血清型鑒定,共鑒定出72株豬鏈球菌2型(占53.3%),14株7型(占10.4%),14株9型(占10.4%),2株1型(占1.5%),33株其他型(占24.4%)。這表明在我國河南及其周邊省份的豬群中,鏈球菌為分離率最高的細(xì)菌性病原,其中豬鏈球菌仍占重要地位(占43.5%),在豬鏈球菌中,血清2型占主導(dǎo)地位(占53.3%)。2.2011~2015年間,豬源鏈球菌的分離率介于23.5~45.5%之間,其中2011年和2014年顯著偏高(p0.01)。另外發(fā)現(xiàn)其分離率具有明顯的季節(jié)性,其中2~5月和8月份顯著偏高(p0.01)。在分離出鏈球菌的772份病料中,有274份僅分離出鏈球菌,沒有發(fā)現(xiàn)其他常見病原菌,占35.5%;有498份同時(shí)分離出各種不同的共感染菌,占64.5%。鏈球菌最常見的共感染菌是副豬嗜血桿菌占所有共感染病料的47.4%;(236/498),其次是大腸埃希菌(35.3%;176/498)、多殺性巴氏桿菌20.9%;(104/498)、支氣管敗血波氏桿菌(15.3%;76/498)、葡萄球菌(6.2%;31/498)、沙門氏菌(6.1%;30/498)和胸膜肺炎放線桿菌(1.0%;5/498)。這表明在豬鏈球菌病的發(fā)生具有明顯的季節(jié)性,且與其他病原菌的共感染情況非常嚴(yán)重。3.對分離鑒定的49株豬鏈球菌,包括2型26株、7型14株、9型9株,和實(shí)驗(yàn)室前期分離鑒定的2株C群馬鏈球菌獸疫亞種強(qiáng)毒菌株進(jìn)行了小鼠毒力試驗(yàn),結(jié)果表明C群馬鏈球菌獸疫亞種的菌株毒力表現(xiàn)最強(qiáng),其次是豬鏈球菌2型、7型和9型菌株。分別選擇小鼠毒力試驗(yàn)中表現(xiàn)最強(qiáng)的豬鏈球菌2型3株、7型3株、9型3株和2株C群馬鏈球菌獸疫亞種菌株(共11株鏈球菌),進(jìn)一步進(jìn)行仔豬毒力試驗(yàn),結(jié)果表明2株C群馬鏈球菌獸疫亞種菌株和3株豬鏈球菌2型菌株的攻毒仔豬均能出現(xiàn)典型豬鏈球菌病的臨床癥狀和病理變化,3株豬鏈球菌9型菌株中有1株(SS9-6)能導(dǎo)致攻毒仔豬出現(xiàn)典型臨床癥狀和病理變化;但是3株豬鏈球菌7型菌株均不能。這表明在我國豬鏈球菌病的病原中,C群馬鏈球菌獸疫亞種和豬鏈球菌2型菌株的致病力較強(qiáng),而豬鏈球菌7型和9型較弱。另外,各血清型豬鏈球菌的小鼠毒力試驗(yàn)結(jié)果和仔豬毒力試驗(yàn)結(jié)果并不完全一致。4.以C群馬鏈球菌獸疫亞種M1株和豬鏈球菌2型SS2-2株為代表菌株進(jìn)行了培養(yǎng)基篩選試驗(yàn)。在含10%新生牛血清的TSA培養(yǎng)基、牛血清瓊脂培養(yǎng)、鮮血瓊脂培養(yǎng)基基、麥康凱瓊脂培養(yǎng)基和巧克力瓊脂培養(yǎng)基共5種固體培養(yǎng)基中,2株菌株均在含10%新生牛血清的TSA中生長最快。在含10%新生牛血清的TSB培養(yǎng)基、馬丁肉湯培養(yǎng)基、硫乙醇酸鹽培養(yǎng)基(TM)、THB和BHI共5種液體培養(yǎng)基中,2株菌株均在含10%新生牛血清的TSB中生長活菌含量最高。對上述的11株鏈球菌進(jìn)一步研究了其在TSA固體培養(yǎng)基上的培養(yǎng)試驗(yàn),結(jié)果表明,豬鏈球菌2型、7型、9型菌株之間的生長速度無明顯差異,其24h培養(yǎng)菌落的直徑介于1.62~1.88 mm之間,但C群菌株的生長速度顯著偏快(p0.01),其平均直徑為3.24 mm。在TSB液體培養(yǎng)基中的生長曲線測定結(jié)果表明,11株鏈球菌的最高活菌含量介于8.4×108~1.2×109 CFU/mL之間,均能高密度發(fā)酵培養(yǎng)。21種生化反應(yīng)的試驗(yàn)比較結(jié)果表明,11株鏈球菌的生化反應(yīng)均不一致,即使同一血清型的不同菌株之間也具有較大差別。藥敏試驗(yàn)表明,11個(gè)強(qiáng)毒菌株對青霉素G、氨芐西林、阿莫西林、頭孢噻肟、頭孢拉定、頭孢克洛、阿奇霉素等均高度敏感。這些試驗(yàn)篩選出了適合豬源鏈球菌培養(yǎng)的最佳培養(yǎng)基,同時(shí)比較了11株強(qiáng)毒菌株的生化特征、生長特性、藥物敏感性等生物學(xué)特性,為后期滅活疫苗研發(fā)奠定基礎(chǔ)。
[Abstract]:In recent years, Streptococcus suis are widely popular in China and has become one of the main pathogens, pathogenic bacteria and serious harm to the development of pig industry. The epidemiology of swine streptococcus, the virulence characteristics, biological characteristics were studied, the main results are as follows: 1. through the isolation and identification of Streptococcus suis (2014.09~2015.12), and the identification results of the laboratory separation for statistical analysis (2011.01~2014.08), the results showed that during 2011~2015, identified 772 strains isolated from Henan and the surrounding area the size of farms and backyard farmers 2521 incidence of swine lung diseases such as material organization (Streptococcus), the total separation rate of 30.6%. by specific the PCR method based on GDH gene of the 310 strains of Streptococcus suis were identified using PCR method, we identified 135 strains of Streptococcus suis (S.suis), accounting for 43.5%. based on CPS gene The specific PCR method was used to identify the serotype of 135 strains of Streptococcus suis were identified in 72 strains of Streptococcus suis type 2 (53.3%), 14 strains of type 7 (10.4%), 14 strains of type 9 (10.4%), 2 strains of type 1 (1.5%), 33 strains of other type (24.4%). Show in Henan and its surrounding provinces in herds, streptococcus was the highest isolation rate of bacterial pathogens, including Streptococcus suis still occupies an important position (43.5%), in Streptococcus suis serotype 2, the dominant (53.3%).2.2011~2015 years, between the separation rate between 23.5~45.5% swine streptococcus, which in 2011 and in 2014 was significantly higher (P0.01). Also found that the separation rate has obvious seasonal, which 2~5 in April and August were significantly higher (P0.01). In the separation of 772 cases of Streptococcus, 274 only isolated Streptococcus, not found in other common pathogens, accounted for 35.5%; 498 copies and isolated different Co infection of bacteria, accounted for the most common 64.5%. co infection of streptococcus bacteria Haemophilus parasuis disease material co infection accounted for all 47.4%; (236/498), followed by Escherichia coli (35.3%; 176/498), Pasteurella multocida (20.9%; 104/498), b.bronchiseptica (15.3%; 76/498). Staphylococcus (6.2%; 31/498), Salmonella (6.1%; 30/498) and Actinobacillus pleuropneumoniae (1%; 5/498). This shows that in the highly seasonal occurrence of Streptococcus suis disease, and other pathogenic bacteria infection is very serious on the identification of.3. from 49 strains of Streptococcus suis type 2, including 26 strains, 14 strains of type 7, type 9 and 9 strains of lab isolated 2 strains of group C Streptococcus equi subsp.zooepidemicus virulent strain of mouse virulence test, the results showed that C of Streptococcus equi subspecies Gunma strains showed the highest virulence, followed by Streptococcus suis type 2, type 7 and type 9 choose small strain. In the performance of the strongest virulence test of 3 strains of Streptococcus suis type 2, type 7 and 3 strains, 3 strains of type 9 and 2 strains of group C Streptococcus equi subsp.zooepidemicus strains (11 strains), further piglets toxicity test, clinical symptoms and pathological changes showed that the 2 strains of group C Streptococcus equi subsp. Strains and 3 strains of Streptococcus suis type 2 strains of inoculated piglets were typical Streptococcus, 3 strains of Streptococcus suis type 9 strains in 1 strains (SS9-6) can cause the inoculated piglets showed typical clinical symptoms and pathological changes; but 3 strains of Streptococcus suis serotype 7 strains were not. This shows that the Streptococcus suis disease in China the pathogen, Streptococcus equi subspecies C strong horses and the pathogenicity of Streptococcus suis type 2 strains, and 7 Streptococcus suis type 9 and weak. In addition, mouse virulence test results of various serotypes of Streptococcus suis in piglets and virulence test results are not entirely consistent with the.4. C group streptococcus beast NDV M1 strain and subspecies of Streptococcus suis serotype 2 strain SS2-2 as the representative of the culture medium screening test strains. Cultured in TSA containing 10% fetal calf serum, bovine serum agar, blood agar base, Jimmy Kang Kai agar and chocolate agar 5 solid medium, 2 strains were in TSA containing 10% fetal calf serum in the culture medium. The fastest growth in TSB containing 10% fetal calf serum, Martin broth, thioglycollate medium (TM), THB and BHI 5 kinds of liquid medium, 2 strains were in TSB containing 10% fetal calf serum in growth the content of the highest living bacteria. 11 strains of Streptococcus further research the culture test in TSA solid medium showed that Streptococcus suis type 2, type 7, no significant difference in growth rate between the 9 strains, the 24h culture colony diameter ranged from 1.62~1.88 mm, but C strains of the students Long too fast speed significantly (P0.01), the average diameter of 3.24 mm. growth curve in TSB liquid medium were tested. The results show that among the 11 strains of the highest viable content ranging from 8.4 * 108~1.2 * 109 CFU/mL, were able to test high density fermentation.21 biochemical reactions and the comparison results showed that the biochemical Reaction 11 strains are inconsistent, even between different strains of the same serotype has great difference. The drug sensitivity test showed that 11 virulent strain of penicillin G, amoxicillin, ampicillin, cefotaxime, Cefradine, cefaclor, spectinomycin, were highly sensitive. These tests selected the best medium for cultivation of swine streptococcus, and compare the biochemical characteristics of 11 strains of virulent strains of growth characteristics, biological characteristics of drug sensitivity, laying the foundation for the later development of the inactivated vaccine.

【學(xué)位授予單位】：河南科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S852.611

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