山羊流产胎儿绵羊附睾种布鲁菌(Brucella ovis)的分离和鉴定

从山羊流产胎儿中分离出1株绵羊附睾种布鲁菌(Brucella ovis),经培养发现该菌需要CO2,不产生H2S,且经硫堇、碱性复红和A、M、R因子血清凝集试验及噬菌体裂解试验等方法鉴定,该菌株与国际标准菌株绵羊附睾种特性完全一致。在山羊中分离出绵羊附睾种布鲁菌株(Br.ovis)在国内尚属首次发现。     

鸵鸟新城疫病毒和大肠埃希菌感染的诊断

从某鸵鸟养殖场发病死亡的鸵鸟肝脏和脾脏用鸡胚分离到一株病毒,经血凝试验和血凝抑制试验鉴定为新城疫病毒,动物致病性试验表明该病毒对雏鸡有较强的致病性.同时分离到一种细菌,通过培养特性、菌体形态、菌落形态、染色特性及生化试验等一系列鉴定,确定为大肠埃希菌.药敏试验表明该株大肠埃希菌对头孢类抗生素高度敏感,而对红霉素、氨苄青霉素、四环素等抗生素表现为不同程度的耐药.     

一株A型副鸡嗜血杆菌的分离鉴定

从山东省某蛋鸡场发病鸡群中分离到一株细菌,经培养特性观察、生化试验、V因子生长试验、致病性试验和PCR试验,证实分离菌为一株致病性副鸡嗜血杆菌,血清型鉴定该菌为A型。药物敏感性试验表明分离菌对头孢噻肟、头孢曲松、丁胺卡那霉素高度敏感,对硫酸新霉素、强力霉素中度敏感,对磺胺间甲氧嘧啶钠、环丙沙星、庆大霉素耐药。     

禽源布利丹沙门菌的鉴定及耐药性分析

本试验从临床送检的病料中分离得到3株革兰阴性杆菌,对分离菌进行分离培养,观察生长特性,进行生化特性和药物敏感性鉴定,对其进行了invA鉴定后确定为沙门菌,后对其进行沙门菌血清型鉴定,鉴定结果表明,该菌为布利丹沙门菌,血清型模式为O9,12:H:g,m,q。药敏试验显示,分离株对庆大霉素、头孢噻肟、舒巴坦、左氧氟沙星、卡那霉素、丁胺卡那霉素、氟苯尼考有较高的敏感性,对多粘菌素B、链霉素有较高的耐药性。     

猪源缓慢葡萄球菌的分离鉴定及致病性研究

某猪场发生一种高发病率、高死亡率的传染病。本研究从该场濒死猪只的心脏血液及肺脏中分离出1株细菌,经镜检、菌落形态观察、培养特性观察及法国生物梅里埃API鉴定系统鉴定,确定该分离菌株为缓慢葡萄球菌。药敏试验表明该菌对多种药物具耐药性,但对氟哌酸、红霉素和强力霉素等敏感;致病性试验表明,分离菌可以引起实验小鼠的慢性死亡,猪只腹腔注射可复制病例,表明缓慢葡萄球菌具有致病性并很可能会对公共卫生安全具有潜在威胁。     

14型副猪嗜血杆菌的分离鉴定及其致病性检测

为探究河南省某大型养猪场育肥猪发生疫情的原因,本研究从该猪场采集的病料样品中进行细菌分离,并对分离菌经革兰氏染色及电镜观察其形态、16S rRNA PCR鉴定;采用多重PCR鉴定其血清型并将扩增的funAB基因与GenBank中的其它副猪嗜血杆菌(HPS)参考株的相应基因进行同源性分析并构建该基因的进化树;采用纸片扩散法鉴定分离菌株的药敏特性并利用小鼠进行了其致病性试验。结果显示:经形态观察、培养特性鉴定以及16S rRNA的PCR鉴定,该分离菌为HPS;经多重PCR并经测序鉴定分离菌为14型HPS,将其命名为JZ-2019;funAB基因的同源性分析及进化树结果显示,JZ-2019株与14型HPS IA-84-22113株(KC795520.1)的同源性为100%。药敏试验结果显示,分离菌对β-内酰胺类、大环内酯类以及氨基糖苷类药物敏感,其中对大环内酯类药物最为敏感;小鼠致病性试验结果显示,小鼠均出现临床症状,甚至死亡,且小鼠各脏器组织均出现不同程度的出血等剖检症状;经PCR扩增从死亡小鼠的脑、腹水中扩增出与分离菌株一致的细菌。表明,该分离菌对小鼠有较强的致病性,且能通过小鼠血脑屏障。本研究证明了河南省猪群中存在14型HPS的感染,为中原地区防控HPS病提供了参考依据。     

乌鳢舒氏气单胞菌的分离鉴定及药敏试验

对广东省佛山市某乌鳢养殖鱼场患病的乌鳢进行病原分离,将疑似分离菌S-1株进行形态和培养特性、生化特性鉴定、PCR鉴定、药敏试验和人工感染等试验。结果表明,该分离菌为舒氏气单胞菌。药敏试验结果表明,本分离菌对壮观霉素、米诺环素、庆大霉素、氧氟沙星、诺氟沙星、头孢类等多种药物敏感。本研究为舒氏气单胞菌病细菌学检测及治疗提供了科学依据。     

致羔羊腹泻大肠埃希茵的分离鉴定及防治

从陕西省某羊场发生腹泻羔羊的肝脏和脾脏分离到一株细菌,通过培养特性、茵体形态、菌落特征、染色特性、生化试验等一系列的系统鉴定,确定为致病性大肠埃希菌。动物致病性试验表明,该菌对小白鼠有较强的致病性,引起其死亡。毒素试验表明,分离菌产生致小白鼠死亡的外毒素。血清学鉴定该茵血清型为O114。药敏试验表明,该茵对大观霉素、头孢噻肟等高度敏感,对米诺环素、克林霉素、克拉霉素等不敏感。分离茵制成铝胶佐剂灭活苗免疫羊群,能有效的预防该羊场羔羊腹泻的发生。     

猪链球菌2型的分离鉴定及免疫预防

从某猪场分离到一株细菌,通过培养特性、菌体形态、菌落形态、染色特性、生化试验及血清学试验等一系列的系统鉴定,确定为猪链球菌2型。动物致病性试验表明该菌对小白鼠和仔猪有较强的致病性,引起其死亡。药敏试验证明该菌对青霉素、氯霉素等高度敏感,对头孢拉定、卡那霉素等中度敏感,对氟派酸等不敏感。分离菌制成油佐剂灭活苗能有效的预防该猪场猪群的发病。     

牛病毒性腹泻病毒四川株SC的分离鉴定及其生物学特性分析

为分离鉴定牛病毒性腹泻病毒(BVDV)四川流行毒株,并掌握其生物学特性和遗传特征,通过病毒分离培养特征观察、RT-PCR检测、间接免疫荧光试验、中和试验4种方法分离鉴定出1株BVDV,命名为SC株。生物学特性检测显示,该毒株对乙醚、氯仿、胰蛋白酶敏感,耐碱不耐酸,对温度敏感,属于RNA病毒;5'UTR遗传进化表明显示该毒株与BVDV-1b型同源性较高。本研究成功分离鉴定1株BVDV四川流行毒株,为BVDV感染机制的研究、疫苗的研制以及防控措施的制定奠定了基础。     

Serodiagnosis of Histophilus ovis-associated epididymitis in rams

An ELISA for the detection of antibodies to Histophilus ovis was used to evaluate the association of epididymal lesions in rams with serologic response to His ovis. Comparison of ELISA results for His ovis in groups of rams with epididymal lesions with ELISA results of clinically normal rams (control group) revealed a significant difference (P less than 0.01) between the control group and those rams from which His ovis was isolated. A significant difference (P less than 0.01) was noticed between the control group and rams with lesions from which an organism other than His ovis or Brucella ovis was isolated. Additionally, a significant difference (P less than 0.01) was noticed in ELISA results between the control group and affected rams from which no organism was recovered and in which the epididymal lesion was not limited to the head of the epididymis. A difference was not detected in the His ovis ELISA results between control rams and rams with lesions associated with a B ovis infection or rams from which no organism was recovered and in which the epididymal lesion was limited to the head of the epididymis. The serologic findings in our study suggest that His ovis is more important in the development of epididymitis in rams than culture results alone would indicate.     

Development of serum antibody activity as determined by enzyme-linked immunosorbent assay to Psoroptes ovis (Acarina:Psoroptidae) antigens in cattle infested with P. ovis

Calves infested with Psoroptes ovis (Hering, 1838) for the first time (naive) or previously infested calves were examined for serum antibody activity to P. ovis (obtained from rabbits) antigens by enzyme-linked immunosorbent assay (ELISA) to determine the temporal appearance of specific serum antibody activity. The development of the serum antibody activity to P. ovis antigens was then correlated with the development of lesions (% scab) and with changes in the number of P. ovis. The serum antibody activity [ELISA OD414 value (EODV) greater than 0.290] to P. ovis antigens in naive calves, in most cases, is first detected at the same time or slightly after the detection of mites and mite-caused lesions; and the development of specific serum antibody activity paralleled the increase in the P. ovis population and the percentage scab until 7 weeks post-infestation (PI). In previously infested calves, if serum antibody activity to P. ovis antigens was not already present from the previous infestation, specific serum antibody activity was detected at the same time and developed in a similar manner as in the naive calves. The serum antibody activity to P. ovis antigens could be detected after calves were relieved of their P. ovis burdens by pesticide treatment or after the P. ovis population began to decline when the calves were allowed to groom themselves. Serum antibody activity to P. ovis antigens was not detected in any of the control calves during the test period.     

Evaluation of electrophoretic immunoblotting for Brucella ovis infection in deer using ram and deer serum

AIMS: Recently the first case of natural infection of deer with Brucella ovis was discovered. The aim of this study was to develop and evaluate an electrophoretic immunoblotting method for testing deer serum for specific B. ovis antibodies. METHODS: An existing immunoblotting method for sheep serum was altered by using a recombinant protein G-alkaline phosphatase conjugate and Tris-buffered saline containing 3% non-fat dry milk powder for the blocking step and the serum and conjugate dilutions. The method was evaluated using 106 sheep sera from B. ovis - negative, accredited flocks, 69 sera from chronically infected rams shedding B. ovis in their semen, 110 sera from a B. ovis-infected flock, 18 sera from stags from which B. ovis was isolated, and 48 sera from deer flocks free from B. ovis infections. The immunoblotting method was applied to another 85 deer sera. RESULTS: The sensitivity of the new immunoblotting method was 98.6% for sheep and 94.4% for deer, and the specificity 99.1% for sheep and 100% for deer. Sixty-nine out of 97 deer sera, originating from the property from which the first B. ovis deer case had been reported, tested positive or suspicious in the complement fixation test. Of these, 53 sera exhibited staining patterns in blots typical for B. ovis infections and also one serum which was negative in the CFT. Only six out of 1498 deer sera. from throughout New Zealand had positive or suspicious reactions in the B. ovis complement fixation test. Of these, one exhibited a staining pattern in the blot suggestive of a B. ovis infection, while four showed patterns of suspicious reactions. CONCLUSION: The new immunoblotting technique is useful as a confirmatory serological test method for B. ovis infections in deer.     

SEROLOGICAL COMPARISON BETWEEN HISTOPHILUS OVIS, ACTINOBACILLUS SEMINIS AND BRUCELLA OVIS

SUMMARY: The serological relationships between 4 strains of Histophilus ovis , the neotype strain of Actinobacillus seminis and Brucella ovis were examined using a cross-absorption complement-fixation technique.
It was found that the 4 strains of H. ovis were serologically homologous and that an incomplete relationship existed between these organisms and A. seminis. Anteriserums prepared against one strain of H. ovis and the strain of A. seminis gave a weak, apparently nonspecific cross-reaction with Br. ovis antigen.
The practical significance of these results is discussed.     

Isolation and identification of Babesia ovis in Extremadura (Spain).

A morphological and morphometric comparison was made of a Babesia ovis strain isolated in Extremadura (Spain) and a B. ovis strain from Turkey. Strains were morphologically similar, with a clear predominance of single sub-spherical or anaplasmoid forms. The use of an image analyser to measure B. ovis merozoites revealed significant differences in the long axis, short axis and volume between the two strains. Both strains however behaved in a similar manner in the indirect fluorescent antibody (IFA) test, which suggests a high degree of common antigenicity, despite morphometric differences. Conversely, no cross-reaction was observed between these strains of B. ovis and the other antigens used: B. motasi (The Netherlands), B. motasi (Turkey), B. crassa (Iran) and Theileria ovis (Turkey). B. ovis is therefore considered to be the aetiological agent of babesiosis in the area under study.     

羊附红细胞体病PCR检测方法的建立

根据羊附红细胞体的16S rRNA基因参考序列，设计1对特异性引物．建立了检测羊附红细胞体的PCR技术。用本方法从感染血样中特异扩增出1条预期大小为1169bp的片段。该方法灵敏、快速、特异性高．可用于羊附红细胞体病的早期快速诊断和流行病学调查。     

A modified indirect immunofluorescent assay for the detection of antibody to Eperythrozoon ovis in sheep

Experimental ovine eperythrozoonosis was studied using Giemsa staining of blood films and a modified indirect immunofluorescent antibody assay (IFAA). The serums of 21 Border Leicester Merino cross lambs between 12 weeks and 7 months-of-age were analysed before and after infection with Eperythrozoon ovis (E. ovis) using the IFAA test. No rise in the IFAA titre was seen until day 7 and this coincided with the first detection of E. ovis organisms in blood smears stained with Giemsa. The percentage of E. ovis infected red blood cells peaked on day 14, but the IFAA titre did not peak until day 35. Titres to E. ovis, on average, had begun to drop by day 63. There was considerable individual variation in response to E. ovis infection as measured by the IFAA. Titres as high as 6,400 were observed in individual sheep at the peak of E. ovis parasitaemia of red cells. One sheep had a titre of 51,200 nineteen days after infection, and titres of 3,000 were maintained for several months in a few sheep. The assay proved reliable, and up to 100 samples per day could be tested. The antigenicity of the slide preparations was found to be satisfactory after storage for 6 months at -20 degrees C and 4 degrees C and for 28 months at -70 degrees C. Temperature fluctuations during storage rendered slides unsuitable for the IFAA after these times. A method of storing E. ovis infected blood in liquid nitrogen is described.     

Effects of vaginal Brucella ovis infection of red deer hinds on reproductive performance, and venereal transmission to stags

AIMS: To investigate the effects of vaginal Brucella ovis infection on the reproductive performance of red deer (Cervus elaphus) hinds. To determine whether stags may become infected with B. ovis by venereal transmission from mating infected hinds. METHODS: Thirty mixed-age red deer hinds serologically negative for B. ovis antibodies were synchronised for oestrus on 22 March 2000. B. ovis was inoculated into the vagina of each hind at oestrus and again, 18 days later. At oestrus, hinds were randomly allocated to six groups, each joined with a 16 month-old red deer stag seronegative for B. ovis, for 55 days. Hinds were blood sampled and scanned for pregnancy using rectal ultrasonography at monthly intervals. Six pregnant and four non-pregnant hinds were slaughtered pre-calving and three hinds were slaughtered post-calving. Reproductive tracts and foetuses were examined grossly, histologically and microbiologically. Calves were identified and blood sampled within 3 days of birth. Hinds and calves were blood sampled in February and May 2001 and vaginal swabs were collected from hinds for B. ovis culture. Blood was collected from stags, 5 and 19 days after mating and semen was collected for B. ovis culture. The 17 remaining hinds were mated in 2001 to two mixed-age wapiti (Cervus canadensis) stags. Both stags were blood sampled after mating. Sera were tested in a B. ovis complement fixation test (CFT) and enzyme-linked immunosorbent assay (ELISA). RESULTS: All 30 hinds developed B. ovis antibody levels, measurable using either the CFT or ELISA, but these did not remain elevated. There was no evidence of infection, either by gross pathology, histopathology or microbiological culture in the ten hinds or six foetuses slaughtered pre-calving. All remaining 20 hinds produced normal calves, 15 of which survived until weaning. Three hinds experienced dystocia and gave birth to dead calves and two calves died within 4 days of birth. One hind which had dystocia was euthanased. Samples from this hind and from 3/5 dead calves showed no evidence of B. ovis infection. B. ovis was cultured from the vagina of 1/19 hinds 48 weeks after inoculation, at which time B. ovis CFT and ELISA results for this hind were negative. Most calves had B. ovis serum antibodies at 1-3 days of age but levels were negligible when sampled at 10-15 weeks of age. Foetuses and dead calves were all seronegative. Three of the five red deer stags used for mating became infected with B. ovis. The two wapiti stags used to mate the remaining 17 hinds the following year remained seronegative. CONCLUSIONS: B. ovis is unlikely to have significant detrimental effects on the reproductive performance of red deer hinds. Venereal transmission via the vagina of hinds is a possible route of transmission between stags. It is possible that survival of the organism in the vagina of some hinds could create difficulties in disease control programmes. CLINICAL RELEVANCE: B. ovis infection of hinds at the time of mating is unlikely to cause significant reproductive losses. Venereal transmission of B. ovis between stags via the hinds may occur when groups of hinds are joined with more than one stag.     

Transmission of Brucella ovis from rams to red deer stags

AIM: To determine whether B. ovis will transmit from infected rams to non-infected red deer stags (Cervus elaphus) grazing together in the same paddock. METHODS: Six rams artificially infected with B. ovis were grazed with six non-infected 14-month-old red deer stags for a four and a half month period from March 4 to July 20, 1999. Stags were blood sampled at one- to six-weekly intervals to test for B. ovis antibodies using a complement fixation test. Stags that seroconverted were semen sampled to test for B. ovis infection by bacteriological culture. RESULTS: Between day 92 and day 124 of grazing together (June 4 and July 6), sera from five of the six stags became positive in the B. ovis complement fixation test. B. ovis was cultured from semen samples from four of the seropositive stags. CONCLUSIONS: Brucella ovis can be transmitted from infected rams to non-infected stags grazing in the same paddock, suggesting that B. ovis infection in farmed deer in New Zealand initially came from infected rams. Whether transmission occurs from direct contact between rams or stags, or indirectly by environmental contamination needs to be established.     

Meningoencephalitis and other conditions associated with Histophilus ovis infection in sheep

Histophilus ovis was isolated from 29 sheep in 20 flocks and 2 artificial insemination (AI) centres in southern New South Wales from 1984 to 1990. The clinical and pathological findings were consistent with previous reports and included polyarthritis (7 flocks), epididymo-orchitis (5), meningoencephalitis (3), pneumonia (3), septicaemia (2), mastitis (1) and metritis (1). Six sheep had meningoencephalitis, a syndrome not previously associated with H ovis infection in sheep, which was similar pathologically to thromboembolic meningoencephalitis in cattle, caused by the related organism, Haemophilus somnus. H ovis was isolated from the semen of 12-month-old rams in a flock that had polyarthritis due to H ovis, in 4-month-old ram lambs and from the uterus of a ewe in a flock that had sporadic cases of H ovis septicaemia.